Ohio State University

NIH Research Projects · FY 2026 · 2021-12

PROJECT SUMMARY/ABSTRACT Melanoma is more prevalent in men than women, suggesting sex hormones may influence this disease. Clinical studies correlate decreased estrogen receptor beta (ERβ) expression with disease progression. However, the mechanisms by which the receptor protects against melanoma formation and progression remain unknown. Our preliminary data show that ERβ loss accelerates tumor formation in a murine melanoma model thereby confirming the tumor suppressor activity implicated in the clinical data. The melanocyte ERβ cistrome overlaps with key melanocyte transcription factors that act as master regulators of differentiation, proliferation, and migration. Estrogen-regulated genes in melanocytes are associated with differentiation and migration pathways supporting a co-regulatory link between ERβ and these master regulators. In addition to the tumor suppressor function of ERβ in melanocytes, ERβ has a melanocyte-nonautonomous function that results in reduced immune infiltrates within the tumor. Furthermore, an ERβ-specific agonist can activate T cells, reduce immune checkpoint inhibitor expression, and increase T cell activation. These data lead to the overarching hypothesis that ERβ activity represses melanoma initiation and progression by modulating melanocyte-intrinsic master regulator activity and enhancing immune responses to the tumor. In this proposal, the hypothesis will be tested by 1) Defining the melanocyte-intrinsic ERβ activities that repress melanoma onset and progression; 2) Determining the influence of ERβ-regulated immune activities on melanoma initiation and therapeutic response.

NIH Research Projects · FY 2026 · 2021-12

Project Summary Sudden, unexpected death in epilepsy (SUDEP) is a fatal complication, occurring in patients with epilepsy, who were otherwise healthy. Dravet syndrome (DS), which results from a loss of function of the sodium channel (NaV) isoform NaV1.1, is associated with a particularly high risk of SUDEP. Notably, the unexpected mortality of SUDEP mirrors cases of sudden cardiac death (SCD) – unexpected death resulting from cardiac arrhythmias. Emerging evidence points to physiological commonalities between the brain and the heart, suggesting that the very molecular mechanisms that underlie epileptic disorders could also directly impact the heart, leading to life- threatening cardiac arrhythmias and thereby, SCD. However, the precise mechanisms by which DS directly impact the heart to promote cardiac arrhythmias and SCD remain unknown. Thus, we propose to utilize models of DS to address this gap in knowledge and to explore the role of direct cardiac remodeling in SCD. Using these models, we will test the hypothesis that defects in NaVs that underlie inherited epileptic disorders, such as Dravet syndrome, dysregulate cardiac Na+/Ca2+ cycling, thereby promoting life-threatening arrhythmias and contributing to SCD. Thus, we propose to: 1) Define the phenotypic and structural impacts of tissue-specific expression of DS-associated defects. 2) Determine the functional impact of DS-associated Na+/Ca2+ signaling nanodomain remodeling on cardiac Ca2+ handling. 3) Assess efficacy of targeting Na+/Ca2+ signaling nanodomains for SCD prevention. Information gleaned from these studies will be used to develop new therapeutic approaches to treat SCD in DS.

NIH Research Projects · FY 2026 · 2021-12

Humans are generally social and these social relationships can promote a healthier lifestyle and increase longevity. Yet, when these social relationships are lost or come to an end, the perceived social isolation and accompanying loneliness can lead to significant physiological, psychological, and social consequences that impair an individual’s ability to function and greatly impairs their quality of life. Although it is widely recognized that social bonds contribute to health, the mechanism(s) by which social bond disruption translates into compromised mental and physical health are not fully appreciated. An understanding of the biological substrates that drive social bonds, and the negative sequelae following loss of social bonds, is essential to develop a framework for pharmacological interventions to reduce health risks associated with loneliness. The conditions and disorders associated with loneliness have each been linked to inflammation and metabolic disruption. The proposed studies will determine the extent to which social bond disruption engages inflammatory and metabolic substrates in the brain of a social species. Given that the negative effects of loneliness are not simply ameliorated by the presence of other individuals in humans, it is essential to examine the underlying protective mechanisms of social bonds, and the biology engaged when bonds are broken, in order to develop interventions aimed at optimizing physical and psychological outcomes for those lacking positive social support. The overarching hypothesis of this line of inquiry is that the neural-glial response engaged by social bond disruption increases neuroinflammation and compromises neural mitochondrial function through disruptions in oxytocin (OT) signaling. Using the socially and genetically monogamous California mouse (Peromyscus californicus), we will determine the extent to which pair bond dissolution increases the neuroinflammatory response to challenge and the extent to which stress-related behaviors can predict the anticipated inflammatory response to pair bond dissolution (Aim 1). We will also test whether pair bond dissolution disrupts synaptosome mitochondrial function (Aim 2). Lastly, we will determine to what extent OT can ameliorate the impact of pair bond dissolution on the neural-glial axis (Aim 3). The impact of social experiences on the brain may be critical to understanding the biological drivers of mental health disorders and neurodegenerative conditions, like Alzheimer’s Disease and related dementias.

NIH Research Projects · FY 2024 · 2021-12

Project Summary: Cochlear implants (CIs) are an effective treatment for adults with severe-to-profound sensorineural hearing loss, and ideally lead to improved real-world communication skills and social engagement. However, we currently do not have a good understanding of relevant, modifiable factors that contribute to the enormous variability in communication outcomes observed in both new and experienced CI users. Further, the current method by which we quantify CI outcomes, via clinical measures of speech recognition, tells us little about the real-world functioning of our CI patients. Our long-term goal is to better understand real-world communication, learning, and adaptation in adult CI users, and the sensory and cognitive-linguistic processes that support maximizing real-world CI benefits, in order to identify future targets for rehabilitative efforts. New CI users largely learn to adapt to their CIs through their everyday experiences. Differences in the real-world auditory- social experience of new CI users likely contribute to communication outcomes, but currently our knowledge of how real-world experience promotes adaptation in adult CI users is limited. Further, the extent to which auditory- social experience changes following implantation remains unclear. Therefore, the overall objectives of the proposed research are to determine how diverse auditory-social experience facilitates speech recognition abilities in new CI users, and to characterize changes in their auditory-social experience, using a multi- disciplinary approach. Here, we focus on the amount and variability of new CI users’ speech input (“speech input attributes”), through assessment of their social networks and communication practices. Our central hypothesis is that speech input variability experienced in the real-world auditory-social environment promotes robust speech recognition abilities in new CI users, and that access to diverse real-world auditory-social experience expands as a benefit of implantation. Our hypothesis will be tested in two Specific Aims: Aim 1 will determine the contribution of speech input attributes experienced in the real-world auditory-social environment to robust speech recognition abilities in new CI users. We will assess the association between new CI users’ experience of speech input variability and amount and speech recognition abilities in challenging conditions, pre- operatively and over 6 months of CI use. Aim 2 will investigate changes in the real-world auditory-social experience of new CI users following implantation, and the moderating role of robust communication skills. We will compare speech input attributes pre-operatively and at 6 months of CI use, and evaluate the relation between changes and speech comprehension ability. The proposed research will lead to a better understanding of modifiable factors (i.e. auditory-social environments) that best support adaptation and will help to define real- world social engagement benefits of implantation. Findings will provide a scientific basis for the development of new rehabilitation protocols and clinical assessment tools to optimize real-world outcomes in CI patients.

NIH Research Projects · FY 2025 · 2021-11

Renewal of my Midcareer Investigator Award in Patient-Oriented Research (K24) will allow me to continue my strong efforts directed toward successful original investigations and mentoring to generate research capacity in mobile technologies to improve health (mHealth). The impact of the initial K24 on my mentees has been striking, with nearly two dozen of my mentees being PI on NIH funded studies (under K, R, or Loan Repayment Program mechanisms). The impact the K24 on my own research has been substantial. Since 2014, I have been Principal Investigator (PI) on six federally funded awards: 1) one newly funded 4-year R01 from NIDA; 2) one R56 from NIDA; 3) one two-year R01 from NIDA; 4) one 4-year R01 from NIDA; 5) my K24 from NIDA; and 6) one newly funded 4-year DARPA research award. In addition, I received a William J Fulbright Foreign Scholarship to Malaysia. As my K24 evolved, so has my research focus. My overall research objective is to devote sufficient time to investigate use of next generation (“NextGen”) technologies, invisible biosensing, and machine learning in substance abuse research. My first research project, Virya, uses “digital phenotyping” with invisible biosensing and machine learning, to identify acute pain exacerbations in individuals on opioids, a population at risk for problematic opioid use. Finally, the R01-funded MyTPill project compares the NextGen technology of unobtrusive ingestible biosensors that provide vivid, indisputable measures of medication ingestion. NextGen research discoveries become commercial products, but this process of introducing new discoveries into the healthcare economy remains deeply unfamiliar—yet increasingly important— to academic clinician-scientists. The objective of my Career Development Activities, therefore, involves learning how to navigate regulatory processes, FDA approvals, valuations, venture capital, contracts and intellectual property rights. Because of the growing importance of commercialization of research findings to the health and sustainability of academic careers, my overall mentoring objective of this K24 is to develop researchers who not only have the skills to conduct rigorous NextGen mHealth studies, but also to extend the mass of NIH-funded mHealth research into the healthcare economy. My mentoring approach incorporates several components that contributed to the success of my initial K24: dedicated funding for mentee’s early investigations; outstanding institutional support; several sources of referral of high-quality mentees; and renowned co- mentors who will help me establish a pipeline of mentees that have risen to that “cutting edge” where successful academic careers begin. This proposed K24 renewal, with its focus on NextGen technologies, invisible biosensing, and machine learning, is highly relevant to NIH’s mission because the lessons learned from this substance abuse research are directly applicable to the care of patients with common, intractable, and expensive conditions.

NIH Research Projects · FY 2025 · 2021-09

This project develops, estimates, and simulates a cutting-edge model of the health and economic impacts of infectious diseases and policy responses to them. The model has four original features: First, it allows for two-way interactions between infections and economic outcomes. These are important because even though infectious diseases are fundamentally a matter of public health, it is essential to account for how diseases and the policy responses to them affect the economy and how, in turn, the economic impacts affect health. Second, it builds policies into a model that accounts for geosocial spread because infectious diseases are spread through contact with others and policy decisions in one area affect the rest of the country. Third, it allows for incidence rates that are measured only through imperfect proxies, which is particularly important for modeling the prevalence of an infectious disease. Lastly, the model accounts for differences in impacts across demographic groups (e.g., by age). This work extends earlier pilot projects that develop a model with the first and second features. Once complete, the model will make it possible to identify the best sets of economic and health outcomes, including infections and mortality, that can been achieved and the policies that would produce those best-case scenario outcomes. It will also make it possible to rigorously quantify the health and economic costs of deviating from optimal policies. The model, which will focus on the United States, can also be applied to cross-country analysis and its features are intended to apply to public health crises more generally, such as the opioid epidemic. The ability to apply the model to a range of epidemics will allow policy makers to compare and contrast the impacts of different epidemics as well as the same epidemic in different locations using a common approach. Additionally, the project conducts several less structured analyses that document the health and labor market impacts of infectious diseases and the policy responses to them. This work will generate results of interest in their own right as policy makers weigh and measure the efficacy of public health responses, will help identify key phenomena to incorporate into our model, and will help us to ensure that the qualitative simulation results are robust to a range of plausible parameter estimates.

NIH Research Projects · FY 2025 · 2021-09

Imbalance contributes to falls, fall risk increases with age, and falls cause accidental death and injury. While causes of falls and imbalance are multifactorial, modern multivariate statistical analyses allow us to quantify the impact of age-related vestibular declines on falls as well as the impact of age-related vestibular declines on age- related balance declines. For example, we have recently reported that nearly 50% of cross-sectional age-related balance declines found using a standard Romberg test variant (“eyes closed; standing on foam”) are mediated by an age-related increase in roll tilt thresholds, suggesting that a single sub-clinical facet of vestibular function is a substantial contributor to age-related balance declines. Furthermore, an analysis of over 5,000 Americans showed that individuals who failed to complete this same balance test condition had significantly increased odds to have reported “difficulty with falling” in the past 12 months (odds ratio of 6.3). We propose to build upon these findings to quantify the multiple links between aging, vestibular function, balance, and falls. We posit the following links: (i) vestibular function declines with age, (ii) these age-related vestibular declines impact balance, (iii) imbalance contributes to falls, and (iv) falls cause death and injuries. Specifically, we posit that correlations between aging, vestibular function, and balance arise, in part, because vestibular imprecision (i.e., vestibular thresholds or vestibular “noise”) increases with age as demonstrated by large increases in vestibular thresholds above the age of 40. Increased vestibular imprecision, in turn, contributes to instability, which contributes to falls. To quantify these correlations and hypothesized causal links, we propose comprehensive assays of vestibular function, broad multi-faceted balance assays, as well as reports of falls. We also propose to further develop and test candidate interventions designed to improve tilt thresholds (“vestibular precision”), which could also improve balance and reduce falls. Given life-or-death significance and broad impact, we propose: Aim 1: To quantify the links between age, vestibular function, balance, and falls in healthy individuals using a comprehensive vestibular function test battery, broad multi-faceted balance tests, & fall-related surveys. Aim 2: To quantify both tilt thresholds and balance before and after a training intervention that has subjects performing a standard tilt threshold task, except that feedback (“correct” or ”incorrect”) is provided after each trial. This task is designed to improve vestibular thresholds (“precision”), which may also improve balance. For Aim 2A, we propose to test young adults (age 18-40). For Aim 2B, we propose to test older adults (age 65-89). Aim 1 will quantify links between vestibular function and falls for the first time and will also quantify new links between vestibular function and balance. This will define potential vestibular and/or balance tests that can be used as screening tests to determine who would benefit from intervention. Aim 2 will quantify the impacts of a training paradigm designed to reduce tilt thresholds. This will develop and test an intervention designed to improve vestibular precision, which could also improve balance and reduce falls.

NIH Research Projects · FY 2025 · 2021-09

PROJECT ABSTRACT/SUMMARY. Lung cancer is a disease of older adults, who make up the least studied population of patients in cancer research. Evidence gaps persist in understanding the impact of newer treatments such as immunotherapy on functional status, the ability to recover from disability (resiliency), and clinical outcomes (e.g. symptom burden and treatment toxicity) among older adults. Current evidence indicates that the majority of older adults with cancer prioritize maintaining functional independence (i.e. no disability) over survival. Despite this common desire among older adults, disability in basic activities of daily living (ADLs), instrumental activities of daily living (IADLs), and mobility is a common consequence of cancer treatment. Functional status in terms of ADLs, IADLs, and mobility is often unmonitored throughout the treatment course. Among older adults with lung cancer, poor physical capability and psychological symptoms such as anxiety and depression are common and represent potentially modifiable risk factors to achieve resiliency rather than disability. Interventions that focus on helping older adults enhance resiliency and prevent functional decline during treatment are urgently needed. Therefore, the objective of this application is to test a novel supportive care intervention specifically designed to enhance resiliency among older adults with advanced lung cancer. This supportive care intervention, called Resiliency among Older Adults Receiving Lung Cancer Treatment (ROAR-LCT), is designed to help patients engage in behavior change to preserve functional status, by optimizing a resilience response to cancer treatment, and is delivered virtually to decrease treatment burden for older adults. The first aim of this proposal is to conduct the ROAR-LCT pilot randomized clinical trial, comprising physical therapy plus relaxation versus standard-of-care, to evaluate a) feasibility and b) the preliminary effect on functional status (primary outcome). The second and third aims are to compare secondary outcome differences (physical capability and psychological symptoms) and exploratory outcome differences (symptom burden and treatment toxicity) between the two study groups. As a geriatric thoracic oncologist, my overall career goal is to become an independent physician-scientist conducting research to understand the problem of disability following a lung cancer diagnosis and developing empirically supported strategies to improve a resilience response and prevent disability among older adults. This award and training proposal will provide the support to advance my knowledge of patient-reported functional outcomes for older adults, implement this knowledge in the design of behavioral clinical trials, and share this knowledge in new leadership roles. To help ensure the success of this application, I have developed relationships with a distinguished mentorship team having expertise in behavioral interventions, cancer control, lung cancer, and geriatrics. This award will provide me with the additional training and career development I need to become an independent physician-scientist focused on cancer and aging and to achieve my long-term goal of becoming an international leader in geriatric oncology and a role model and mentor for women in academic medicine.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY There is a global upsurge of falls in older adults that impacts nearly every family across the world. Millions of older adults fall each year in the United States, leading to catastrophic injuries, deaths and soaring healthcare costs. Over the last decade, 911 fall calls have tripled while transport rates to the hospital after a fall have significantly decreased. Instead, 911 is increasingly used for lift assists (falls that do not result in transport). Deployment of emergency medical services for lift assists diverts care from higher acuity emergencies and costs more than 200 million dollars annually in the United States. There is a potentially powerful yet underutilized solution if we leverage the hidden opportunities of fall events, such as lift assists that do not result in catastrophic consequences, to activate prevention strategies. This study aims to develop a scalable strategy for early identification of individuals at high risk of falls and activate prevention solutions. We hypothesize that a systematic 911 fall call intake which has a broader concept of frailty, Frailty And Cognition+Environment (FaCE), will better account for the compounding and cascading nature of fall risks in older adults. At the completion of this project a scalable machine learning model which incorporates FaCE factors to predict high utilization of 911 for falls will be developed. In addition, we will characterize barriers and facilitators for adoption, implementation, and maintenance of fall prevention strategies in the home for patients with FaCE risk factors. This project will utilize a blend of systems science and community-based participatory research approaches and state of the art predictive analytics to elucidate the FaCE of falls, develop a scalable fall prevention solution that can be implemented nationwide and inform a larger-scale implementation trial for using 911 fall calls to activate effective fall prevention strategies in homes.

NIH Research Projects · FY 2025 · 2021-09

Liquid Biopsy for Rapid Detection and Real Time Monitoring of FGFR-altered Cancers Patients with advanced cancers driven by fibroblast growth factor receptor (FGFR) alterations, including gene fusions and single nucleotide variants (SNVs), are benefiting from several new FGFR kinase inhibitors. Erdafitinib and pemigatinib were recently FDA approved for bladder cancer and cholangiocarcinoma, respectively, and other FGFR inhibitors have received fast-track designation and are being explored in tumor agnostic basket trials. Patients are experiencing improved overall survival and progression free survival, with high overall response rates. Unfortunately, virtually all patients eventually develop resistance to these inhibitors, oftentimes through acquisition of secondary FGFR mutations. FGFR inhibitor development is hindered by the lack of accurate and comprehensive methods to 1) rapidly detect FGFR alterations in order to qualify patients for clinical trials, 2) monitor therapeutic response, and 3) characterize emerging drug resistance. The development of novel testing strategies, such as non-invasive liquid biopsies, can fulfill these unmet needs for diagnosis, prognosis, and therapy selection. Liquid biopsies evaluate a blood sample, from which cell-free DNA (cfDNA) shed by the tumor is sequenced to detect genomic alterations. The technical specifications for existing commercial cfDNA tests show that these assays are not validated for FGFR fusions and have insufficient sensitivities of ~30% for FGFR fusions. Additionally, commercial tests are too large and costly to repeat frequently for therapy and disease monitoring. Thus, we propose to develop and validate an FGFR-focused, accurate, and cost-effective cfDNA sequencing assay (FGFR-Dx) for real-time testing to support rapid detection, response monitoring, and early detection of resistance. Our team at Ohio State University has six active FGFR inhibitor clinical trials, a large cohort of FGFR true positives, and a Clinical Laboratory Improvement Amendments (CLIA)-compliant Cancer Genomics Lab with extensive experience performing clinical-grade tumor sequencing and bioinformatics analysis for detection and interpretation of gene fusions and single nucleotide variants. Further, we have paired this cfDNA development with a rapid research autopsy study that will enable the first systematic evaluation of detection limits for cfDNA by assessing how accurately the heterogeneity observed across tumor samples from multiple sites is represented in cfDNA. We propose the following Aims to address the criteria for PAR-18-317: 1) Analytically validate a targeted liquid biopsy assay (FGFR-Dx) to detect fusions and single nucleotide variants (SNVs) in FGFR1-3; 2) Establish the clinical validity of FGFR-Dx to detect FGFR fusions and SNVs. In summary, coupling the development of an FGFR-focused liquid biopsy with rapid research autopsy can broadly impact the field’s understanding and application of cfDNA approaches in patients with FGFR-altered and other cancers.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY/ABSTRACT Individual cells within a given cancer type are capable of expressing a diversity of phenotypic states resulting from an underlying heterogeneity of genetic, epigenetic, transcriptomic, and molecular features. How this diversity evolves and influences therapeutic response is an essential question in cancer biology. One emerging mechanism of developing such heterogeneity is the evolution of microenvironmental niches within tumors that support a cancer stem cell (CSC) state. CSCs are defined by their functional capabilities such as long-term self- renewal, the capacity to give rise to a range of differentiated cell types, and enhanced tumor-forming ability. They are also believed to drive resistance to anti-tumor therapies, such as radiation therapy. Pancreatic ductal adenocarcinoma (PDAC) has a dismal prognosis, with a 5-year overall survival of <10% and a dire need for novel therapeutic strategies. Radiation is an integral part of PDAC therapy, however not all cancer cells respond. Identifying mechanisms of radioresistance would transform the clinical management of PDAC. Using pancreatic tumor models, our preliminary results suggest that secreted Wnt ligands produced by one cancer cell subset drive a Lgr5+ stem-like state in another cancer cell subset, in essence forming a supportive niche that promotes stemness within pancreatic tumors. In this proposal, I will test the central hypothesis that Wnt-driven cellular phenotypic heterogeneity and stemness promote radiotherapy resistance in pancreatic adenocarcinoma. I will examine the role of pancreatic tumor cell subpopulations in radiation resistance, including the Lgr5+ cells (Aim 1) and the Wnt producing niche (Aim 2). Under Aim 1, I will characterize the cancer stem cell properties of Lgr5+ cells in pancreatic cancer, their relative resistance to radiation therapy, and their role in tumor repopulation. These results will determine whether Lgr5+ cells are CSCs and drivers of radioresistance in established tumors and inform their molecular characteristics, which may provide added means to target these cells. Under Aim 2, I will investigate the targeting of the Wnt-producing niche in combination with radiation. Specifically, I will use genetic or pharmacologic perturbation of the Wnt pathway in combination with radiation and assay tumor response and animal survival. These efforts will test the therapeutic potential of Wnt inhibitors as radiosensitizers in PDAC. Collectively, this work will facilitate basic mechanistic insights into both cellular heterogeneity and radioresistance, with the ultimate goal of translating these discoveries and developing improved treatment strategies for PDAC patients.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY Many groups are investigating why some lung cancer patients respond well to radio- and immuno-therapies and some do not. One variable is tumor hypoxia, and many groups have shown it can significantly inhibit the effectiveness to these therapeutic modalities. Clinical studies have identified hypoxia as an independent prognostic indicator of poor patient outcomes, but even though this connection has been known for decades, no FDA-approved intervention exists to clinically overcome hypoxia. Some investigators have tried to deliver more oxygen to the tumor, but this approach remains constrained due to the poorly formed tumor vasculature. We have taken an innovative approach and asked if we can reduce demand for, rather than increase supply of, oxygen to reduce hypoxia. We have found that the FDA-approved vasorelaxant papaverine (PPV) has an off- target ability to inhibit mitochondrial complex 1, and reduce oxygen consumption rapidly, in low micromolar concentrations in every cell line tested in vitro. We have also shown that PPV can enhance the effectiveness of radiation and immune checkpoint blockade (ICB) in preclinical models of lung and other cancers, without sensitizing well-oxygenated normal tissue. Reducing hypoxia reverses immune privilege, decreases terminally- exhausted T cells, and increases progenitors that are responsive to PD-1 blockade. We have more recently developed new derivatives of PPV that have lost their vasorelaxant capability and increased their duration of action so that they can be improved immuno-sensitizers. We now propose to test the hypothesis that PPV can effectively enhance the radio- and immuno-therapeutic treatment of preclinical models of lung cancer, and that it is feasible to add PPV to standard of care therapy for advanced non-small cell lung cancer (NSCLC). We have examined TCGA databases and found that lung cancer driver mutations in the KEAP1/NRF2 pathway lead to high levels of mitochondrial gene expression that can cause elevated oxygen metabolism contributing to hypoxia. In Aim 1, we will investigate the effects of oncogenic NRF2 activation human and murine cells and model tumors to determine the dependence of these cells on mitochondrial function, how increased oxygen metabolism contributes to tumor hypoxia, and if therapy-refractory tumors are sensitized by PPV or its derivatives. In Aim 2, we will examine the effect of tumor hypoxia on the migration and activation of T-cells in model tumors and how the immune infiltrate changes after reduction of hypoxia with PPV or its derivatives. Finally, in Aim 3 we will perform a phase 1 clinical trial to determine if the addition of PPV is feasible for patients receiving standard of care chemoradiation followed by immunotherapy for advanced NSCLC. We will look for effectiveness in changing tumor oxygenation using paired blood level oxygen determination (BOLD) MRIs, and for changes in immune populations of peripheral blood mononuclear cells. These studies will let us know if, and how, to use PPV or its novel derivatives in future clinical trials for the treatment of NSCLC.

NIH Research Projects · FY 2024 · 2021-09

Title: Determining Age-Dependent Metabolic Changes in Tumors and Their Microenvironment Project science areas: 6 MCB, 8 HIB Project Summary/Abstract: Aging is one of the primary risk factors for cancer and deregulated metabolism is a unifying hallmark for both aging and cancer. Yet, we do not fully understand how ageassociated metabolic changes contribute to cancer initiation and progression. Although many animal models are created to fatefully recapitulate the oncogenic events that lead to cellular transformation and cancer progression, most are carried out in young animals that likely have metabolically different tumor microenvironments and robust immune systems. To this date, few studies have systematically examined how age-dependent alterations of cellular and organismal metabolism impact tumor growth and metastasis. We hypothesize that age-associated metabolic dysregulation will significantly alter cellular metabolism both within tumor cells, as well as in cells within the tumor microenvironment, which may influence cancer progression and response to therapies. We propose a multipronged, single cell approach to characterize and resolve metabolic changes over time, both within the tumor and its microenvironment. Finally, we will assess the impact of environmental factors, such as dietary nutrients and microbiome derived metabolites, on cancer initiation and progression. Collectively, we think our findings will be transformative and will answer key questions of how aging rewires cellular metabolism and how it affects tumor growth and metastasis. We hope to elucidate important questions about cellular plasticity, metabolic heterogeneity and identify metabolic liabilities that can be accentuated by limiting specific nutrients or metabolites in new combinatorial therapies.

NIH Research Projects · FY 2025 · 2021-09

MECHANISMS OF CHROMATIN REGULATION OF TRANSCRIPTION PROJECT SUMMARY Over the past decade, the Poirier lab has worked to contribute to the fields of chromatin biology and transcription regulation by developing a research program that is focused on the mechanisms by which chromatin and nucleosome structural dynamics regulate genome accessiblity to transcription regulatory complexes. We have developed and applied on our own and through collaborations, a wide range of experimental tools that enable mechanistic studies including single molecule fluorscence, single molecule force spectroscopy, ensemble fluorescence, and histone chemical ligations. In combination, we have quantitatively investigated how chromatin regulators including histone post translational modifications (PTMs), PTM binding proteins (readers), chromatin remodelers, linker histones, and transcripton factors function to control genome accessiblity to chromatin regulatory complexes. Building off this work, we propose to investigate two central questions in the fields of chromatin biology and transcription in the coming 5 years: (i) How do pioneer transcription factors target their recognition sites within compact nucleosomes and chromatin, and then facilitate chromatin decompaction? (ii) How do epigenetic regulators function together to synergistically or antagonistically regulate genome accessibility to transcription regulatory complexes? Pioneer factors (PFs) are master regulators of cell differentiation, are correlated with nucleosome depletion, and somehow access their binding sites within compact chromatin that are inaccessible to canonical transcription factors (TFs). Furthermore, PF disfunction is strongly correlated with disease most notably cancer. By combining single molecule and ensemble studies we will directly test distinct mechanisms of PF function, and determine what differentiates PFs from canonical TFs. Furthermore, through collaborative work we will investigate the same PFs in vivo to determine how the PF mechanisms that emerge from our in vitro studies impact their functions in vivo. This first direction will provide key insights into how pioneer factors gain access to their sites within compact chromatin and what differentiates PFs from canonical TFs. Epigenetic regulators including histone PTMs, and their readers and writers are critical to organismal development, aging and numerous diseases including cancer. We and others have investigated these regulators individually, yet how they function in combination remains largely unknown. Leveraging our ability to prepare core histone and most recently linker histones with any combination of PTMs, histone H3 readers, and most recently biochemical quantities of endogenous human SAGA and ATAC complexes, we will use single molecule, ensemble and ES cell-based methods to determine how biologically relevant combinations of these regulators control accessibility to canonical TFs and influence PFs. This second direction will provide a mechanistic and functional foundation for understanding how key epigenetic transcriptional regulators differentially control chromatin dynamics, accessibility and transcription.

NIH Research Projects · FY 2025 · 2021-09

Moderate-severe traumatic brain injury (TBI) results in physical, behavioral, and cognitive impairments that can have a devastating impact on functioning in the community. Comprehensive interdisciplinary inpatient rehabilitation can maximize function and reduce complications. However, clinicians and researchers are unable to answer the question, “Which of the wide range of rehabilitation practices can most effectively advance recovery and improve outcomes?” Due to the complexity of the rehabilitation process and the heterogeneity of the TBI patient population, the standard approach to comparative effectiveness research, the randomized controlled trial, is inadequate. Nevertheless, with growing limitations on healthcare resources and shorter lengths of stay, it is urgent and critical to identify the specific rehabilitation approaches that can optimize outcomes for persons with TBI. A pragmatic, prospective observational study to close this evidence gap is proposed. By leveraging the infrastructure of the National Institute on Disability, Independent Living and Rehabilitation Research’s TBI Model Systems, the largest longitudinal study of moderate-severe TBI in the world, the data needed to compare the effectiveness of different rehabilitation approaches will quickly accumulate. Recent advances in data capture, through electronic medical records (EMR), and in advanced statistical methods provide the avenue by which the complexity of rehabilitation can be scientifically studied. Aim 1: This study aims to leverage EMR technology to ensure data identified as critical to rehabilitation treatment are captured through standardized documentation during the natural course of a patient’s hospitalization. Aim 2: Incorporating the findings of preliminary studies on comparative effectiveness of treatment approaches, this study will aim to evaluate the impact of different approaches to treatment on patient outcomes. It is hypothesized that rehabilitation interventions directly targeting performance of real-life activities (ContextTx) and higher-level functions (AdvTx) will individually and in combination improve community participation at 1-year post-injury, as well as on functional independence at discharge and at 1-year post-injury. Aim 3: Identify time-varying patient and setting factors that can change over the course of rehabilitation and that modify the effects of treatment. Advanced statistical analyses coupled with data capture made feasible by effectively designed EMR documentation from frontline providers will provide the data necessary to identify which treatment approaches are associated with better patient outcomes. This in turn will arm clinical providers with valuable knowledge to design the most effective treatment plans for patients. The findings of this study will further empower clinical operators with the necessary information to advocate and promote evidence-based treatments for TBI recovery to fiscal stewards, credentialing bodies, and regulatory agencies.

NIH Research Projects · FY 2024 · 2021-09

Atherosclerotic cardiovascular disease (ASCVD) is a leading cause of death in the United States; and there is ~2-fold greater risk for CVD events for people living with HIV (PLWH).1,2 The mechanisms underlying increased CVD prevalence in PLWH are not fully understood, but likely involve heightened activation of monocytes and macrophages. We hypothesize that monocytes will be exposed to disparate inflammatory signals during differentiation in PLWH; as a result, macrophages will be functionally and phenotypically different and exacerbate CVD in this population. Our data suggest that alterations in the lipidome may influence inflammation, monocyte activation, and the differentiation of macrophages. Antiretroviral therapy may alter lipid profiles by reducing oxidative phosphorylation (OxPHOS) and fatty acid oxidation (FAO) as a consequence of ART-induced mitochondrial dysfunction. Our studies have identified differences in the lipidomes of HIV- and HIV+ populations, even when traditional lipid panels in these groups were similar22. Biomarkers associated with CVD risk (IL-6, sCD14, TNFR1),10-15 were directly related to fatty acid composition and pro-inflammatory lipid classes in PLWH. We have also identified an expansion of pro-coagulant, vascular homing monocytes in PLWH42-44 and linked monocyte activation to altered lipid profiles19,20,45-47. Aim 1: To identify unique and common phenotypic, transcriptomic, and functional MDM profiles associated with the presence or absence of ASCVD in PLWH and HIV- individuals. 1A: To compare MDM phenotypic, functional, and transcriptomic profiles across our 4 groups. 1B: To sort and differentiate monocyte subsets (based on CD14 and CD16 expression)43,49,50 into MDMs in order to determine whether subset origin determines differential functional and phenotypic outcome. 1C: To sort macrophages from atherosclerotic plaques (carotid endarterectomies) and compare the transcriptional profiles of these cells to the profiles identified in MDMs from our 4 participant groups. Aim 2: To characterize drivers of MDM activation in PLWH and HIV- persons with and without ASCVD. 2A: To characterize the plasma lipid profiles of participants using advanced lipidomics (Lipidyzer) as well as the lipidomic profiles of atherosclerotic plaques collected from carotid endarterectomies. 2B: To explore the consequences of in vivo and in vitro lipid profile modulation on MDM gene expression and functional capabilities; statin treatment will improve the lipidome51 and as a consequence, decrease MDM activation, and in vitro exposure of MDMs to proinflammatory lipids (i.e CERs, SaFAs) will activate these cells. 2C: To determine the drug-specific effects of ART exposure on MDM profiles. Aim3: To elucidate profiles associated with ASCVD in HIV- and HIV+ individuals using a comprehensive, multi-dimensional approach and in vitro pathway inhibitor experiments to explore differential drivers and inhibitors of signaling cascades on MDM transcription and functional profiles.

NIH Research Projects · FY 2024 · 2021-09

PROJECT SUMMARY/ABSTRACT We propose herein a methodology to identify synthetic peptide-based binders to non-canonical structural motifs in RNA. Reagents that selectively target these biologically important motifs would be transformative theranostic tools for study and modulation of RNA-governed biology. The long-term goal of our research program is to develop sequence and context-selective reagents for targeting of any such non-canonical structural motif in lncRNAs. The objective of this application is to synthesize a small library of peptide-derived reagents and quantitatively rank their competence in non-canonical hybridization to defined RNA structures, using novel and robust functional screening methods in vitro and in cell culture. In contrast to the striking progress of synthetic biology at the coding interface, non-canonical targeting remains in development. We hypothesize that progress in this area is limited in a number of ways: 1) a prior focus on targeting Watson-Crick (WC) paired bases rather than non-canonical pairs; 2) an over-emphasis on intercalation-driven binding; 3) lack of exploration of secondary structure in RNA targeting reagents; 4) lack of a unified functional assay to rigorously evaluate binding. We further hypothesize that synthetic binding solutions exist for every non-canonical motif; if these solutions could be found, then non-canonical hybridization could be programmed in the same way that duplex hybridization is programmed. Such an advance would enable precise interrogation of nucleic acid biology with novel chemical tools. The rigor of the prior research lies in the known efficiency of synthetic bases in targeting select non-canonical pairs, as well as preliminary data demonstrating tunable and expansive binding selectivity via backbone and base modification. We will test our central hypothesis and accomplish the overall objective of this application via the following three specific aims: 1) Synthesis and evaluation of bPNAs targeting non-canonical sites via base-triple formation; 2) Enzymatic and functional assays for bPNA-RNA targeting efficacy; 3) In vitro and intracellular bPNA targeting and selectivity for structural motifs in native RNAs.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY/ABSTRACT Our goal is to determine the molecular mechanisms of myosin-3 motors, unique kinase-myosin hybrids in the myosin superfamily, that are involved in fundamental cellular processes including cargo transport and the organization of the actin cytoskeleton in sensory cells. Despite their critical role in human biology, the molecular mechanisms of myosin-3 motors are not well understood. We propose that different modes of regulation together with different kinase substrates and binding partners determine the molecular mechanisms of myosin-3 motors. An integrated and innovative biochemical, biophysical, cell biological and high-resolution structural approach will be used to (i) determine the enzymatic profile and regulation, (ii) the unique structure (iii) and pathways that control myosin-3 function and regulation in vitro and in cells. Collectively, these studies will reveal novel mechanistic insights into the regulation of myosin-3 motors and have broad implications in the understanding of how different myosins are tuned to organize actin networks in cells.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY/ABSTRACT Childhood behavior problems are a top mental and public health priority. Problem behavior such as aggression, defiance, and noncompliance emerge as early as 2 years of age and can persist through adolescence and adulthood leading to long term functional and psychological maladjustment. Children born very premature (VPT; gestational age at birth < 32 weeks) are twice as likely to demonstrate behavioral problems as their peers. Despite the risk and occurrence of behavior problems in the VPT population, early intervention tends to focus on treatment of developmental delays with very few resources for parents to manage behavior issues. Behavioral parent training (BPT) is an effective treatment for families of children with problem behaviors. However, little is known about the use of BPT programs for former VPT infants with their unique medical, developmental, and psychological risk profiles. In addition, system and individual barriers effect parent access and engagement in BPT. In an effort to address tailored needs of these families and service barriers, BPT delivery via digital applications and engagement strategies like telephone coaching calls can provide accessible and tailored approaches to support parents of young children. Currently, there is no effective, accessible BPT intervention for former VPT infants. The purpose of the study, “Parent training for parents of toddlers born very premature: A factorial design to test web delivery and telephone coaching” is to develop a new, accessible and effective form of BPT delivery to address the unmet and unique needs of parents of VPT children. The BPT program proposed in this study is an adaptation of the evidence-based based Chicago Parent Program (CPP) called the ezParent. The study design is a factorial design to test a hybrid approach of digital ezParent and telephone coaching calls. Coaching calls are meant to reinforce parent learning and support tailoring strategies to their child’s developmental level and needs. Parent and child (aged 20-30 months corrected) dyads will be recruited from two Midwest NICU follow up clinics within Chicago, IL and Columbus, OH. Parents (n=220) will be randomized to one of four conditions: (1) ezParent+coach, (2) ezParent, (3) Active Control+coach, or (4) Active Control. We will test the independent and combined effects of ezParent and coaching calls on parent and child outcomes. We will evaluate the differences in ezParent dose and engagement with and without coaching calls. Parent and child data will be assessed at baseline, and 3-, 6- and 12-months post baseline. Providing low intensity, digitally delivered interventions has the potential for influencing engagement and address challenges in service delivery for parents of VPT children at higher risk for behavioral problems and with elevated behavior problems.

NIH Research Projects · FY 2024 · 2021-09

PROJECT SUMMARY The nucleoside reverse transcriptase inhibitors (NRTIs) have potent activities against HIV, but their therapeutic benefit in patients undergoing NRTI therapies is limited by significant adverse drug reactions (ADR), resulting in poor patient compliance and compromised drug efficacy. Our group has recently described an indispensable role of a lysosomal nucleoside transporter ENT3 in lysosomal homeostasis via deletion of ENT3 in mice. Intriguingly, ENT3 KO mice manifest clinical phenotypes closely resembling NRTI ADR. The overall objective of this application is to evaluate ENT3-loss driven lysosomal toxicity as a putative mechanism involved in the chronic adverse sequelae of NRTIs. The central hypothesis of this proposal is that NRTIs that do not interfere with ENT3-supported lysosomal homeostasis or the inclusion of lysosomal signaling agents will minimize the occurrence of NRTI ADR. Aim 1 will evaluate the strategies to avoid NRTI toxicity without compromising drug efficacy. Our working hypothesis for this aim is that disruption of interaction between ENT3 and NRTIs or the inclusion of pharmacological agonists of lysosomal-autophagy pathway will mitigate the onset and severity of NRTI ADR. The preliminary studies that demonstrate the involvement of the cell surface NRTI transporters (e.g., ENT1, CNT) and not the lysosomal ENT3 for NRTI efficacy, the misregulation of the AMPK and mTOR signaling axis in the Ent3-/- mice, and the functional rescue of multi-organ dysfunction in Ent3-/- mice using a pharmacological AMPK activator AICAR; all support this aim. Aim 2 will elucidate the mechanism(s) of occurrence of NRTI-specific ADR signs. Our working hypothesis for this aim is that NRTIs, when present at clinically relevant blood concentrations, will inhibit the ENT3-regulated adult stem cell functions resulting in disruption of tissue repair and regeneration. In addition, we hypothesize that NRTIs will differentially impact the ENT3 function in adult tissues to bring distinct inflammatory, metabolic and degenerative changes that coupled with stem cell alterations, will explain the clinically observed NRTI ADR signs. The preliminary studies that demonstrate the transport of many ADR-producing NRTIs by ENT3, the inhibition of lysosomal adenosine transport by NRTIs and the perturbation of lysosomal recycling of adenosine in Ent3-/- mice leading to adult stem cell exhaustion, tissue inflammation and degeneration, and breaches of mesodermal tissue integrity, which taken together supports this aim. The project will utilize biochemical and molecular approaches, novel ENT3 probes, newly generated ENT3 mouse models, metabolomics, tissue engineering, pharmacophore modeling, synthetic and screening procedures and PKPD to accomplish the goals. The successful completion of the project will provide new insights into the mechanisms of occurrence of NRTI ADR and may have translational benefit for optimizing treatments (such as long-term efficacy, adherence, tolerability, etc.) in patients undergoing NRTI therapies.

NIH Research Projects · FY 2024 · 2021-09

PROJECT SUMMARY Although impressive progress has been made to reduce the burden of heart disease, the illness remains the leading cause of death in the United States. Notably, many types of persistent heart injury can cause pathological remodeling of the heart and progression to heart failure (HF). Our goal is to identify novel molecular pathways responsible for such transition, since targeting them therapeutically presents an exciting opportunity to reverse heart remodeling and improve survival. While transcriptional regulators of progression to HF have been studied extensively, the importance of post-transcriptional regulation, such as chemical modification of messenger RNA, has been overlooked. Recent studies recognized that methylation of mRNA in position N6 of adenosines (m6A) was essential for the heart’s ability to adapt to stress, but the master regulator of this mechanism remains unknown. This proposal is aimed to identify the undiscovered cardiac mediator of m6A-dependent mechanism and elucidate how exactly m6A methylation acts to maintain homeostasis in the heart. After methylation, m6A- modified mRNAs can be recognized by specific mRNA-binding proteins belonging to the YTH domain family (YTHDF), of which YTHDF1 is a key member expressed in the heart. Our preliminary proteomics data from mouse hearts show that YTHDF1 is the only member in its family to interact with the protein Hook homolog 3 (HOOK3), which binds motor proteins to activate cargo movement along the microtubules. Since ribonucleoproteins can use microtubular routes to reach their sites of localized translation, our central hypothesis is that YTHDF1 is critical for maintenance of cardiac homeostasis post-stress by modulating m6A-mRNA translation through its interaction with HOOK3. To test this hypothesis, we have already generated a novel conditional mouse model with cardiomyocyte-specific deletion of YTHDF1. In Aim 1, we will subject control and cardiomyocyte-specific YTHDF1 knockout mice to a pressure-overload model of heart failure to define the requirement of YTHDF1 for cardiac stress adaptation. In Aim 2, we will then perform ribosome profiling to determine the global impact of YTHDF1 on mRNA translation and crosslinking immunoprecipitation assays to identify its key mRNA targets in adult cardiomyocytes. Finally, in Aim 3 we will characterize YTHDF1 interaction with HOOK3 and analyze its functional significance using a variety of exciting molecular biology techniques. Overall, our approach is innovative because it aims to define the entirely novel function of YTHDF1 in the heart. This proposal is also significant because it will fill knowledge gaps about the mechanism for YTHDF1-dependent regulation of m6A-mRNA fate through our newly discovered HOOK3 interaction. The proposed study is also significant because targeting direct effectors of mRNA processing such as YTHDF1 opens new avenues for correcting pathological protein synthesis in the heart. Markedly, such knowledge can offer unique transcription- independent targets for restoring healthy biology of cardiac muscle cells.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY Insufficient sleep, sleep disorders, and resulting problems with health and cognition are increasingly common in the United States. Many sleep disorders may be associated with abnormal sleep homeostasis: an innate regulatory process that balances sleep need, sleep intensity, and sleep amount as a function of prior time spent awake. Sleep homeostasis requires a feedback circuit to maintain the system within defined limits. However, the cellular components and protein signaling pathways of this feedback circuit remain incompletely defined. Our understanding of sleep homeostasis thus far is primarily based on the study of neurons, but I showed that non-neuronal cells (i.e. astrocytes) also play a role. I posit that the homeostatic feedback circuit includes a neuronal waking signal that reflects sleep need and an astroglial integrator of the neuronal waking signal. I propose that the wake-promoting neurotransmitter noradrenaline (NA) is a candidate for the neuronal waking signal that interacts with astrocytes. I further propose that calcium (Ca2+) is the astroglial integrator of sleep need because 1) NA increases astroglial Ca2+ activity and 2) I showed that astroglial Ca2+ plays a role in sleep homeostasis. My overall hypothesis is that wake-promoting neurons increase astroglial Ca2+ signaling during elevated sleep need. I will test this hypothesis in two AIMS: 1) Determine how NA impacts astroglial Ca2+ dynamics before, during, and after sleep deprivation (SD); 2) Determine how sleep loss impacts astroglial protein signaling. For AIM 1, I will use a multifaceted approach to optogenetically inhibit or stimulate NA neurons while imaging Ca2+ dynamics in adjacent astrocytes and recording electroencephalographic brain state activity in freely behaving mice. Optogenetics, Ca2+ imaging, and electroencephalographic recordings will occur simultaneously under baseline conditions and during SD & recovery. Using this multimodal approach, I can temporally register cell-type specific neuronal activity and astroglial Ca2+ dynamics within distinct arousal states in freely behaving mice. For AIM 2, I will determine which astroglial proteins respond to changes in sleep need. I will use ultra-performance liquid chromatography-tandem mass spectrometry to quantify astroglial proteins from rested and SD mice using targeted and untargeted proteomics. Targeted proteomics will include NA- & Ca2+-related signaling proteins as well as synaptic, metabolic, and gap junction proteins because these proteins are implicated in sleep homeostasis. I will also determine the phosphorylation status of these proteins because phosphorylation status changes with sleep need and is an important post-translational modification in astrocytes. Astrocytes will be isolated from brains of wild type mice and mutant mice with reduced astroglial Ca2+ signaling. In this way, I can determine which astroglial proteins are 1) responsive to changes in sleep need and 2) Ca2+-dependent. The proposed studies use innovative methods to define biological substrates of sleep homeostasis. These findings, in turn, will further characterize the contribution of non-neuronal cells in the regulation of sleep-wake behavior and will expand our understanding of physiological and disordered sleep.

NIH Research Projects · FY 2025 · 2021-09

Project Summary Sudden cardiac death is a leading cause of mortality in the United States and often results from cardiac arrhythmias. Mutations in Na+ channels, particularly in their carboxy terminal domains (CTDs), dysregulate beat- to-beat cycling of Na+ and Ca2+, and thereby, precipitate arrhythmias. Similarly, mutations in calmodulin (CaM), which closely regulates these channels, are also linked to arrhythmias. However, the role of Na+ channels in arrhythmogenesis remains unclear. NavCTD mutations, which alter the affinity of Nav CaM, impair fast inactivation and induce proarrhythmic late Nav current (INa). This particularly is evident for Nav isoforms which exhibits the lowest CaM affinity and correspondingly, the largest of late INa magnitude relative to peak INa. Our recent studies indicate an important role for these Nav isoforms in late INa-mediated arrhythmias. However, the arrhythmogenic impact of Nav dysregulation in calmodulin-driven arrhythmias remains unclear. Based on strong preliminary data from our laboratory, we hypothesize that affinity of CaM for Nav will dictate the magnitude of arrhythmogenic late INa. Diminished CaM binding to the NavCTD will increase late INa, while enhanced binding will hasten channel inactivation, mitigating proarrhythmic late INa. Thus, we propose to: 1) Assess the extent and mechanism of NaV dysregulation by CaM. 2) Elucidate the relative contribution of mutant CaM-mediated Nav dysfunction to calmodulinopathy associated arrhythmias. 3) Examine the antiarrhythmic potential of enhancing CaM-Nav interaction. Thus, by understanding calmodulinopathies, we aim to discover approaches to prevent arrhythmias stemming from both aberrant CaM-NaV interaction and abnormal NaV function.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY As a ubiquitous HSP90 paralog in the endoplasmic reticulum (ER), GRP94 plays important roles in protein quality control in the secretory pathway by participating in both the unfolded protein response and the ER-associated protein degradation pathway. My laboratory has demonstrated that GRP94 is a strategically important target for cancer, because it controls multiple key molecular pathways in cell growth, migration, immune tolerance and differentiation, including integrins, TLRs, IGF-II, Wnt co-receptor LRP6, and GARP (or LRRC32). GARP (Glycoprotein A Repetitions Predominant) is responsible for surface docking and activation of latent TGFb and a focus of this proposal We have made significant contributions to this area through immunological and biochemical studies, including: 1) that GARP is an important molecule for cancer immune evasion via regulating multiple cell types (e.g., cancer cells, platelets, regulatory T cells, B cells). 2) We discovered a novel mechanism of TGFb activation from cell surface GARP-TGFb complex via proteolytic cleavage of GARP. 3) GARP has been found to be aberrantly expressed in multiple human cancers to promote oncogenesis via both cancer cell-intrinsic and -extrinsic mechanisms. 4) Preclinical studies suggest that GARP is a novel therapeutic target for cancer immunotherapy. These accomplishments have deepened our conviction that the study of GRP94 and its client network will lead to better understanding of this chaperone biology in cancer and to development of novel cancer therapeutics, alone or in combination with approved immunotherapeutic agents. In the next phase of the study, we will address the hypothesis that GRP94/GARP-targeted therapy applied to multiple vulnerable cancers will overcome immune resistance to checkpoint inhibitors. First, we will determine the roles and molecular mechanism involved in GRP94 regulation of TGFb biogenesis, activation and signaling. This aim will focus on structural analysis of the GRP94-GARP complex, and on resolving mechanisms of GRP94 in folding two other molecules important in regulating TGFb signaling: LRRC33 and LRG1. Second, we will develop novel cancer immunotherapeutic strategies targeting GRP94 and GARP. The goal is to advance the top first-in-class agent(s) among several pre-clinical leads through a milestone-driven strategy. This includes agents to inhibit GARP cleavage, GARP-specific antibodies, drug-like GRP94-selective inhibitors, antibodies against the cell surface GRP94 (ectoGRP94) preferentially expressed on cancer cells, and T cells engineered to express chimeric antigen receptor (CAR) composed of a single-chain antibody against ectoGRP94 fused with T cell signaling motifs (GRP94-CAR-T). Overall, the impact of this study lies in fundamental understanding of GRP94 in regulating the TGFb pathway and in developing promising next generation immunotherapeutic agents.

NIH Research Projects · FY 2025 · 2021-09

In utero exposure to environmental chemicals can disturb neurobehavioral development in both animals and in humans. The pathways linking in utero environmental exposures to neurobehavioral development likely involve exposure-induced changes in the function of neural circuits that support cognitive control and reward processes. We hypothesize that changes in the function of these circuits may act as a pathway between environmental exposure and a range of maladaptive behaviors that commonly emerge in late childhood and adolescence, a period that has been largely understudied with respect to the effects of prenatal exposures on neurodevelopmental outcomes. Such behavioral symptoms include attention problems, substance abuse, and psychotic experiences. This study will: 1) apply novel pattern recognition approaches to identify specific exposure profiles of complex high-dimensional mixtures of prenatal chemical and social exposures and examine how these profiles explain variation in risk for these behavioral symptoms in adolescence; 2) use task and resting state functional MRI (fMRI) to identify how distinct exposure profiles affect circuits that support cognitive control and reinforcement learning; and 3) explore if exposure-induced changes in brain activation and connectivity mediate associations between prenatal exposure profiles and behavioral symptoms in adolescence. Impact: This R01 will integrate advanced pattern recognition methods, a cognitive neuroscience approach and state-of-the-art fMRI techniques to explore brain pathways through which prenatal exposures alter behavior later in adolescence. We will explore circuit-based changes in brain function that may mediate associations between prenatal exposure profiles and symptoms of psychiatric disorders that typically emerge and co-occur in adolescence. By simultaneously studying mixtures of chemical and social stressors as well as profiles of co- occurring symptoms, we will greatly enhance our ability to comprehensively characterize the complex impacts of prenatal environmental exposures on behavioral symptoms in adolescence. Our findings will allow public health interventions to improve the quality of children's perinatal environment and the development of novel circuit-specific intervention tools.