Vanderbilt University Medical Center

NIH Research Projects · FY 2025 · 2022-09

Abstract Individuals with autism experience challenges in developing language that impact their long-term social, academic, and vocational success. Previous research has identified several predictors of language in children with autism, but even when these predictors are considered, a large amount of the variance in language remains unexplained. Importantly, many children with autism still do not develop “useful speech” or language ability, despite receiving interventions that target these previously-identified predictors. Thus, there is a pressing need to identify novel predictors of language in children with autism. It has been proposed that differences in sensory responsiveness may emerge early in life from altered brain states and impact a child's ability to engage with others in their environment, thereby producing “cascading effects” on language acquisition in children affected by autism. If this is the case, intervening upon early sensory responsiveness may translate to improved language outcomes, by influencing brain states and boosting engagement in this population. Although intuitively appealing, given our understanding of the precedence and possible “foundational” nature of early sensory development relative to the emergence of “higher-level” language skill, evidence for the cascading effects theory to date has been rather limited. A primary challenge to establishing the aforementioned links is that autism cannot always be reliably diagnosed in the earliest stages of life (i.e., in infancy and toddlerhood). A potential solution is to prospectively follow infants and toddlers known to be at high likelihood for a future diagnosis of autism and language disorder. The Sensory Project in Infant/Toddler Siblings of Children with Autism (Project SPIS) scales up a prior NIDCD-funded R21 (R21DC016144; PI: Woynaroski) in a comprehensive test of the cascading effects framework in infant and toddler siblings of children with autism (Sibs-AUT), approximately one-third of whom will go on to be diagnosed with autism or language disorder, and infants at relatively lower, general population- level likelihood of these conditions (younger siblings of typically developing children; Sibs-TD). NOVELTY AND IMPACT: This innovative and interdisciplinary project is expected to further elucidate the nature of longitudinal links between a potentially tractable, but relatively understudied factor (early sensory responsiveness) and language in infants at elevated and lower, general population-level likelihood of autism. If our hypotheses are born out, findings of the study will lay the foundation for a clinical trial that will examine the efficacy of a targeted treatment of sensory function for distal effects on language as mediated by more proximal effects on resting brain states and engagement in infants at high likelihood for autism. This work is expected to facilitate earlier identification of language impairments and point towards novel targets of early “preventative” interventions, thereby improving the long-term outcomes of children who are diagnosed with, or at heightened risk for, autism and language disorder.

NIH Research Projects · FY 2025 · 2022-09

The Vanderbilt TBEL Center assembles a multi-disciplinary team of field-specific experts to collaboratively investigate the basic and translational pathways of colonic pre-cancer progression. Our foundational work on two subtypes of colonic pre-cancers, adenomas (ADs) and sessile serrated lesions (SSLs), depicts the early origins of tumorigenesis that are shaped by modulation of the immune microenvironment via neoplastic cells and the microbiota. We have shown that SSLs originate from gastric metaplasia arising from the mucosal surface in a cytotoxic immune microenvironment, whereas ADs arise from stem cell-derived WNT activation at the crypt base. In this center, we will extend our investigation of specific biological mechanisms towards the developmental trajectories of these pre-malignant lesions into progression or indolence. Basic Project 1 investigates the contribution of neutrophil-AD crosstalk, largely via dipeptidase 1 (DPEP1) both at the cell surface and released in small extracellular vesicles, in the course of AD progression. Translational Project 2 investigates, in human prospective studies, the association of pks+ Escherichia coli that induces genotoxic stress with pre-cancer progression, as well as colon epithelial cell and mucosa mechanisms that may contribute to a polyp-promoting microenvironment. Basic Project 3 investigates acquisition of stemness in modulating antigen presentation to cytotoxic T cells in the context of co-evolution between neoplastic cells and the immune system. Joint analysis of common colorectal pre-cancer tissues will facilitate an ongoing process of iteration and integration across all projects. Our TBEL Center offers a complementary blend, from reductionist and systems biology approaches, to investigate critical factors involved in the progression of pre-cancerous tumors of the colon to CRC. The work will utilize cutting-edge technologies on human tissues, including single-cell and spatial transcriptomics, small extracellular vesicle profiling, multiplex imaging, longitudinal data analysis, and next-generation computational algorithms. In addition, substantial human polyp resources previously established by the Vanderbilt GI Specialized Programs of Research Excellence and the NCI Moonshot Human Tumor Atlas Network will be leveraged by the same team of investigators in the TBEL Center. In addition, an innovative co-culture system will be employed by each project, where polarizing pre-cancer organoids can be co-cultured with key microenvironment elements exposed to neoplastic cells from the luminal or basal side. This work will inform the modeling of tumor development trajectories and identify mechanisms of progression that will enable improvements in risk stratification, precision prevention, and interception for individuals with colorectal pre- cancers.

NIH Research Projects · FY 2025 · 2022-09

This proposal responds to BRAIN Initiative RFA-NS-20-006 and aims to elucidate the neural and biophysical mechanisms of noninvasive focused ultrasound (FUS) neuromodulation. FUS overcomes shortcomings of other neuromodulation methods and can noninvasively stimulate millimeter-scale regions in any part of the brain including deep brain structures. We seek to understand how different doses and spatiotemporal applications of FUS interact with the brain at cellular, circuit, and behavioral level. When used in conjunction with MRI, the FUS beam can be precisely localized while network-level effects can be observed with BOLD fMRI. In the past few years, through BRAIN Initiative funded projects, we have developed an integrated MRI guided FUS system (MRgFUS) with image-guidance and MRI capabilities required to place the beam accurately in the brain and map its location using magnetic resonance acoustic radiation force imaging (MR- ARFI). Using this system, we have demonstrated that FUS exerts bidirectional (excitatory and inhibitory) and state dependent neuromodulation of the nonhuman primate (NHP) sensorimotor system. FUS directly excites somatosensory area 3a/3b neurons at resting state but suppresses activated neurons when they are engaged in processing tactile inputs and elicits activation in downstream off-target brain regions. Here, we seek to investigate the mechanisms underlying FUS neuromodulation by evaluating neural signals at multiple scales using multiunit array electrodes and functional MRI during FUS neuromodulation over a parameter space chosen to test the influence of pulse duration, pulse repetition frequency, and amplitude. The planned studies will use optogenetics and pharmacological manipulations to test the hypothesis that increasing repetition frequency independent from other parameters preferentially drives specific groups of neurons. Studies varying amplitude will assess a hypothesis derived from our recent observation that FUS at moderate pressures elicits stronger inhibitory effects than high pressures. We will map invasive electrophysiological measurements to non-invasive measurements of neural activity (e.g. BOLD fMRI) that can be used in humans. Safety will be assessed with imaging, deep learning analysis of hand grasping behavior, and post-mortem assessment, providing important information for the ongoing translation of FUS neuromodulation. Our proposed studies will elucidate mechanisms underlying FUS neuromodulation over a broad parameter space in experimental models that span the individual neurons through whole brain networks and connect these multi scale electrophysiological and functional MRI observations made at the cellular, local microcircuit, and global levels to behavior changes in a NHP model system. Thus, our approach is closely aligned with the goals of this RFA and will further our knowledge of FUS neuromodulation, a fast-growing non-invasive method for dissecting circuits in the mammalian brain that offers the potential for therapeutic interventions to diseases involving abnormality in regional and network functions.

NIH Research Projects · FY 2026 · 2022-09

Alzheimer’s disease (AD) is the most common type of dementia and according to the Alzheimer’s Association, it currently affects approximately 7.2 million Americans age 65 and older. Even more striking, AD prevalence is expected to increase to nearly 13 million Americans by 2050. Unfortunately, despite this high prevalence, the molecular and cellular mechanisms underlying the onset of AD remains to be fully elucidated. Vascular risk factors, such as hypertension, are becoming increasingly recognized as key contributors to dementia. Hypertension affects nearly half the adult population in the United Stats, making it one of the most prevalent yet modifiable risk factors for AD and related dementias (ADRDs). Therefore, unraveling the molecular and cellular mechanisms by which hypertension leads to ADRDs will contribute to finding novel therapeutic targets with the potential to prevent and/or ameliorate cognitive deficits. Matrix Metalloproteinase-9 (MMP-9) is a molecular player associated with learning and memory and is dysregulated in both hypertension and dementia. Therefore, the purpose of my K00 project is to understand the molecular and cellular mechanisms that link hypertension to the onset of dementia through MMP-9 dysregulation in the vasculature. My research will focus on the role of MMP-9 activity in modulating cerebral blood flow (CBF), neuronal activity, and cognitive impairment in a mouse model of hypertension. Overall, the central hypothesis is that hypertension leads to increased MMP-9 activity, which will impair CBF regulation and neuronal activity in the retrosplenial cortex (RSC), contributing to impaired neuronal activity during demanding mnemonic tasks such as associative memory. In addition to providing me with a unique training opportunity, my three aims will utilize a repertoire of state-of-the-art techniques and translational data from participants, and its successful completion has the potential to highly impact the field of neurodegeneration by uncovering potential novel disease-modifying therapeutic targets to prevent and ameliorate cognitive deficits associated with hypertension.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT Higher prevalence of sepsis in older individuals is also linked to increased rate of diagnoses for multiple types of dementia including Alzheimer’s disease and related dementias (ADRD). Chronic impairments include changes in memory and attention, emotional function and neuromuscular strength which each have a major impact on patients and families. Exact mechanisms underlying this persistent damage are unknown but data support a role for chronically activated microglia following infection. Understanding of neurological changes during this critical time period is hampered by a lack of appropriate animal models and short experimental endpoints. Behavioral assessments can be severely confounded by hypoactive delirious states and sickness behaviors, and many studies have been conducted in young rather than aged animals, without additional AD-relevant neuropathology further limiting interpretive and predictive power. We will address each of these challenges and test the overarching hypothesis that overarching hypothesis that age and AD neuropathology predispose the brain to greater neuronal damage, prolonged microglial activation and poorer cognitive outcomes following sepsis. Further, we hypothesize that microglia of older adults have a more robust inflammatory response and a reduced ability to downregulate inflammation leading to persistent cognitive impairment, particularly in the presence of additional AD-relevant neuropathology. Our three aims will be conducted in young adult (3 months) and aged mice (up to 18 months), and in vivo data will be supported by ex vivo electrophysiology, immunohistochemistry and molecular biology approaches. We will assess our key outcomes up to 90 days following recovery from illness. In Aim 1 we will demonstrate the extent of increased risk for persistent cognitive impairment and prolonged neuroinflammation following sepsis in aged compared to young animals. We will use a cecal slurry injection to induce sepsis in mice up to 18 months using a rodent model that recapitulates the major defining features of acute brain dysfunction including: acute neuroinflammatory response, disrupted EEG patterns, behavioral deficits across a range of cognitive and neuromuscular domains, and impaired hippocampal long-term potentiation. In Aim 2 we will directly examine the role of microglia in persistent cognitive impairment following sepsis using clinically relevant biomarkers, immunohistochemistry morphological quantification of microglia and whole tissue and single cell RNA approaches. In Aim 3 we will determine the extent to which AD neuropathology contributes to sepsis- induced cognitive impairment and brain dysfunction. We will compare behavioral outcomes and microglial response in two different mouse models of AD to test the potential for increased sensitivity to CS in the presence of common AD neuropathology and gene variants.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT Pulmonary arterial hypertension (PAH) is an orphan disease with a delayed diagnosis and markedly elevated mortality from right heart failure. Despite nearly a dozen FDA-approved drugs for PAH, median survival is only seven years. All approved therapies target one of three vasodilatory pathways, and none are disease modifying. This application has two objectives: 1) Understand dynamic and static relationships between molecular markers and PAH progression and resilience; 2) Identify molecular features of PAH risk and resilience in individuals harboring a PAH-causing mutation. It is unknown why some at risk individuals develop PAH and others do not. BMPR2 mutations are present in about 30% of patients with PAH but clinical penetrance is only 20%. Unaffected BMPR2 mutation carriers (UMCs) are a unique and understudied population that may also provide clues to disease trajectory in patients with clinical PAH. Longitudinal natural history studies with molecular profiling in PAH are lacking. Most molecular profiling studies in PAH are cross- sectional which limits understanding of how disease progression and disease markers relate over time. We propose a strategy of dense clinical and molecular phenotyping at multiple timepoints to overcome inferential limitations of cross-sectional studies. This application will leverage the clinical and research infrastructure built at Vanderbilt over the past 35 years in our study of PAH patients. The investigators share an extensive published record of recruiting patients with this rare disease and related UMCs. We hypothesize that a comprehensive understanding of risk and resilience over time in patients and genetically susceptible individuals will provide insight into disease severity and identify novel therapeutic targets in patients with PAH. Aim 1 will identify static and dynamic molecular features of disease progression and resilience. 1a: Perform serial clinical, proteomic, and gene expression profiling in HPAH, IPAH, and healthy controls 3 times over 4 years. Bioinformatic and network medicine analyses will identify proteins and RNAs associated with changes in clinical outcomes, functional capacity, and RV function in the parent cohort and two external validation cohorts. 1b: Test whether adding molecular risk/resilience markers will improve the performance of a widely used PAH risk prediction tool (REVEAL 2.0 Risk Score). Aim 2 will identify the clinical and molecular factors that promote resilience and susceptibility to PAH in a longitudinal cohort of UMCs. UMCs will undergo serial clinical and molecular phenotyping as in Aim 1. Proteins/genes that mirror PAH are “risk factors” and those that mirror a healthy population are “resilience factors”. Explanatory models will be developed and tested in validation cohorts. We will test UMC risk and resilience features for associations with clinical outcomes in PAH patients and risk prediction performance. These studies will identify signatures of risk and resilience to PAH progression and penetrance, offering an initial step toward personalizing care and surveillance guided by biologic data.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY The last two decades have seen extraordinary advances in the cost, accessibility, and interpretability of genetic testing. In the context of this astonishing progress, it is striking that for many rare genetic diseases, diagnostic delay – the time between onset of symptoms and a diagnosis – has not improved. Current health care services are unable to effectively identify patients that would benefit most from genetic testing. As a result, many patients affected by genetic disease are not diagnosed for years after symptoms develop, or are never diagnosed at all, leading to costly diagnostic odysseys, health care disparities in genetic services, and preventable morbidity and mortality for those with conditions that have an effective, targeted treatment. Much of what we know about genetic disease is based on studies of individuals and their families. This has proven to be a powerful method for discerning the clinical characteristics of genetic disease, generating one of the most enduring and useful resources in medical genetics: the online Mendelian inheritance in man (OMIM). However, clinical descriptions in OMIM do not always match the way diseases are described in real- world EHR data. To improve our ability to use genetic testing effectively, we can learn, at scale, from the data clinically captured while testing and diagnosing patients. EHRs provides an opportunity to study genetic disease from a new perspective, enabling scalable methods that augment existing the knowledge base to include phenotypes observed in real-world health care data. This proposal builds on our prior work curating genetic testing data from the EHR and developing tools to identify undiagnosed patients from characteristic genetic disease profiles. Specifically, we have built a database of clinical genetic testing information extracted from the EHR for over 20,000 individuals, with detailed information regarding test results, variant interpretation, and diagnosis. From this resource, we can define EHR-based cases series of individuals with genetically-confirmed clinical diagnoses of genetic disease. We will use a data-driven approach to discern characteristic phenotypes from the EHR-based case series, and merge these results with clinical descriptions from OMIM. This approach seeks to translate the curated, durable knowledge cataloged in OMIM to a portable and scalable product that can layered on any set of EHRs to identify undiagnosed patients with genetic disease. The ultimate goals of this proposal are leverage these data and tools to 1) translate and add to clinical curations of genetic diseases using real world EHR data, 2) assess diagnostic yield of EHR-based tools that identify undiagnosed patients and 3) characterize the contribution of demographic and phenotypic features that lead to earlier or later diagnosis.

NIH Research Projects · FY 2025 · 2022-09

Hundreds of variants in the COL4A3, COL4A4 and COL4A5 genes cause a broad range of glomerulopathies affecting the function of the glomerular basement membrane (GBM) in patients with Alport syndrome. These genes encode the assembly of collagen IV α345 scaffolds, the major constituent of the GBM. The pathogenic variants lead to a broad array of clinical manifestations, ranging from microscopic hematuria to end stage renal disease. The underlying mechanisms linking these variants with GBM abnormalities and renal failure remain obscure. Current therapy is limited to treatment with ACE inhibitors to slow progression and new therapies are in urgent need. How genetic variants of the α345 scaffold cause Alport syndrome remains unknown. Here, we focus on Z-variant causing Alport syndrome without loss of the α345 scaffold but rather reduction- or loss-of-function effect (hypomorph variant). We use this variant as a vanguard to decipher the pathogenic mechanism of the α345 collagen IV in Alport GBM and develop new forms of therapy. In Aim 1, we will utilize a new Z-variant animal model to understand mechanisms of α345 collagen IV dysfunction. In Aim 2, we will determine exact defects caused by Z-variant and similar pathogenic variants at protein and cellular levels to identify specific targets for small molecule therapies. In Aim 3, we will test protein replacement therapy and screen for pharmacological chaperones correcting assembly and stability of the collagen IV α345 scaffold. The completion of the Aims will advance our knowledge about Alport pathogenesis and lay out foundation for therapy development addressing causative mechanisms in Alport syndrome.

NIH Research Projects · FY 2025 · 2022-09

Chronic kidney disease (CKD) has a worldwide prevalence of 10-15% and claims over 130,000 lives annually in the US. Systemic potassium deficiency hastens progression of CKD while higher dietary potassium is protective; however, the mechanistic link between CKD progression and potassium remains entirely unknown. Coordination of total body potassium homeostasis involves both the kidneys and skeletal muscle, with inwardly rectifying potassium (Kir) channels in both organs being critical molecular regulators of this process. I aim to understand how alterations in plasma potassium determine CKD progression. As multiple cell types are involved in the progression of CKD, my focus is on potassium-mediated signaling on proximal tubule epithelial cells and renal macrophages. Using a combination of whole animal kidney physiology, single cell RNA-seq, electrophysiology, and cell biology, I will determine how Kir channels in muscle, proximal tubule cells, and macrophages mediate kidney injury and if these pathways can be targeted as a novel approach to slow CKD progression. I propose to (1) determine how the basolateral potassium channel, Kir4.2, in the proximal tubule mediates kidney injury and metabolism, (2) determine how Kir2.2 in macrophages affect kidney injury, and (3) determine how Kir2.1 and Kir2.2 in skeletal muscle affect systemic potassium balance and in turn, kidney injury. Successful completion of this proposal will change our fundamental understanding of Kir channels as regulators of potassium signaling and identify Kir4.2, Kir2.1, Kir 2.2, and their related pathways as targets for CKD treatment. Further, it will define a muscle-kidney axis linked by plasma potassium as the signaling molecule. This project is my first as an independent investigator and will be supported by significant institutional support from Vanderbilt University Medical Center. This includes funding, laboratory space, and protected research time to establish my research program focused on potassium signaling via Kir channels and CKD.

NIH Research Projects · FY 2026 · 2022-09

Current understanding of how and why race influences the transition from acute to chronic pain following traumatic injury remains limited. Traumatic injuries result in over 30 million emergency department visits and 2.5 million hospitalizations each year in the U.S. Risk for developing post-injury chronic pain is significantly greater for non-Hispanic Black (NHB) patients compared to non-Hispanic White (NHW) patients with similar injuries. Although NHB patients experience higher levels of acute post-injury pain and are more likely to transition to chronic pain than NHW patients, the biobehavioral and social factors that influence this transition are not well understood. The overall aim of this proposal is to improve understanding of the factors that influence the transition to post-injury chronic pain and shape racial pain disparities. To address our aims, we will recruit 150 NHB and 150 NHW adults with traumatic orthopedic injuries from a level 1 trauma center and assess pain outcomes during hospitalization and across monthly follow-ups. First, we will identity similarities and differences in the extent to which biobehavioral factors explain post-injury chronic pain in NHB and NHW patients. We hypothesize that greater pain sensitivity (assessed in hospital via quantitative sensory testing), elevated acute inflammatory biomarkers (hsCRP, TNFα, IL1β, IL-6), more negative cognitions (catastrophizing, pain/treatment expectations), and higher depressive/posttraumatic stress symptoms will predict post-injury chronic pain (i.e., higher odds of chronic pain onset, greater pain intensity and interference) in both NHB and NHW patients. If chronic pain risks are moderated by race, we hypothesize that worse post-injury chronic pain in NHB relative to NHW patients will be explained, in part, by higher biobehavioral risk factors. Second, we will identify similarities and differences in the extent to which social factors explain post-injury chronic pain in NHB and NHW patients. We hypothesize that social risk factors (greater life and neighborhood stress, lower socioeconomic status) will predict post-injury chronic pain in both NHB and NHW patients, and that social protective factors (higher social support) will buffer against in the influence of social risk factors. If chronic pain risks are moderated by race, we hypothesize that worse post-injury chronic pain in NHB relative to NHW patients will be explained, in part, by higher levels of social risk factors. The comprehensive assessment of risk and protective factors across multiple levels of the biopsychosocial model will advance understanding of the pathways that contribute to post-injury chronic pain for both NHB and NHW adults, including factors implicated in racial differences in transition to chronic pain. Evaluating in-hospital evoked pain sensitivity as a predictor of post-injury chronic pain development represents a major innovation. This knowledge could spur the development of cost-effective, scalable screens for hospital settings to redress racial disparities in pain. The investigators assembled for this multidisciplinary team possess complementary skill sets in traumatic injury, chronic pain, quantitative sensory testing, inflammation, pain-relevant biobehavioral and social factors, and racial disparities in chronic pain.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY This proposal seeks to further develop and validate a cell-size-based MR imaging method dubbed MRI-cytometry as a novel approach for imaging tumor-infiltrating lymphocytes (TILs), and to evaluate its potential as a surrogate imaging biomarker to predict treatment response to immune-checkpoint blockade (ICB) immunotherapy. ICB is the most widespread class of immunotherapies but it poses a new challenge to assess tumor therapeutic response. ICB induces infiltrations of TILs into tumors to kill cancer, but such increased numbers of TILs may lead to transient tumor enlargement, which is the current indicator of tumor progression i.e., non-responders. Therefore, effective response to ICB could be misdiagnosed as tumor progression i.e., pseudo-progression. There is often a much longer waiting period than conventional treatments to verify the persistence of tumor volume changes, which prevents timely adjustments of treatment plans particularly in non-responder patients, causing unwanted treatment delays, costs, and risks of toxicity. Current mainstream imaging methods require exogenous agents for labeling TILs, which significantly increases cost and risks of toxicity. To overcome these limitations, this application proposes a novel, exogenous-agent-free approach for imaging TILs. Because lymphocytes (5-10 μm in diameter with and without activation) are significantly smaller than most cancer cells (10-20 µm), cell size could be used as an endogenous contrast to distinguish “small” TILs from “large” cancer cells. The significantly increased TILs are expected to cause some unique microstructural changes e.g., decreased mean cell sizes and increased cell fractions of “small” cells. To detect these changes, we recently developed a cell-size-based MRI method dubbed MRI-cytometry for imaging cell size distribution and intracellular volume fractions in vivo. Based on our preliminary data, we hypothesize that MRI-cytometry can serve as a specific imaging biomarker of TILs, and hence can predict treatment response to ICB immunotherapy. To test our hypothesis, we propose three specific aims: Aim 1 [development]: To further develop MRI-cytometry imaging to overcome potentially confounding effects for accurate estimation of microstructural parameters. Aim 2 [validation]: To validate MRI-cytometry imaging to characterize lymphocyte infiltration following ICB immunotherapy using preclinical animal models. Aim 3 [evaluation]: To evaluate MRI-cytometry to predict tumor response to combinations of ICB immunotherapy and radiotherapy. Successful completion of this project will establish a new exogenous-agent-free MRI tool for imaging tumor-infiltrating lymphocytes following immune-checkpoint blockade immunotherapy. It can be used as an “add-on” to clinical MRI to reduce cost and risks of toxicity compared with those lymphocyte labeling methods. Moreover, the proposed approach is expected to be translated to the clinic easily, allowing clinicians to stratify responder and non-responder patients early to adjust treatment plans in the manner of personalized medicine.

NIH Research Projects · FY 2025 · 2022-09

Project Summary/Abstract Acute kidney injury and brain injury manifest as delirium each affect over 25% of the 500,000 patients undergoing cardiac surgery annually, and these organ injuries are associated with longer hospitalization, increased mortality, longer mechanical ventilation, but the underlying mechanisms are poorly understood. Vascular reactivity is a component of vascular function which enables control of tissue perfusion via smooth muscle tone. Vascular reactivity is elicited via endothelial nitric oxide synthase signaling which may be disrupted by inflammation, surgery, and oxidative stress. The endothelium is a major regulator of vascular reactivity, coagulation, and immune cell activation, and endothelial dysfunction may contribute to organ dysfunction after surgery. We have recently identified that impaired vascular reactivity is independently associated with kidney injury after surgery. Under normal circumstances, nitric oxide diffuses from the endothelium to vascular smooth muscle and binds to the heme-moiety on soluble guanylyl cyclase inducing a conformational change that allows the enzyme to catalyze the formation of cyclic guanosine monophosphate. Oxidation of the iron in this heme moiety to the ferric state, however, impairs nitric oxide binding and vascular reactivity. Impaired vascular reactivity secondary to impaired soluble guanylyl cyclase activation may be an important mechanism underlying perioperative organ injury that is not routinely addressed in clinical care. Recently, pharmacologic soluble guanylyl cyclase stimulators have been developed to enhance impaired soluble guanylyl cyclase activation and are approved for use in patients with heart failure and pulmonary arterial hypertension. These drugs stabilize the heme moiety on soluble guanylyl cyclase while binding to the enzyme at a separate site, and they directly stimulate soluble guanylyl cyclase to generate cyclic guanosine monophosphate. This project builds on our prior findings that identified impaired vascular reactivity as a potential mechanism for perioperative organ injury, and further examines the role of the soluble guanylyl cyclase stimulator vericiguat in enhancing vascular reactivity in patients undergoing cardiac surgery and at risk of organ injury. We hypothesize that administration of a soluble guanylyl cyclase stimulator prior to surgery will enhance perioperative vascular reactivity and decrease cellular markers of renal tubule and neuronal injury. We will directly assess vascular reactivity in vivo and ex vivo using gold standard techniques in patients randomized to receive vericiguat versus placebo and will compare plasma and urine biomarkers of soluble guanylyl cyclase stimulation, endothelial activation, endothelial barrier degradation, renal tubular injury, and neuronal injury.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT Patients with pulmonary arterial hypertension (PAH) have reduced health related quality of life (HRQOL) and impaired exercise capacity. Despite fourteen approved therapies, most patients die within ten years. This grim fact underscores the need to develop interventions that improve HRQOL, particularly targeting mechanisms that are non-redundant to existing PAH therapies. Increasing physical activity is highly efficacious in PAH, resulting in six-minute walk distance (6MWD) and HRQOL improvement that often exceeds the effect of medications. Prior activity studies required inpatient rehabilitation, which is impractical, hard to sustain, and poorly scalable to a rare disease. Moreover, rehabilitation for PAH is not typically reimbursed by insurance in the United States, highlighting the need for alternatives to promote physical activity. This application builds on a recently-completed, NIH-funded pilot and feasibility trial of a mobile health (mHealth) intervention in PAH. In the pilot, subjects wore a Fitbit device and were randomized to either usual care or a fully automated “smart text” intervention. Text content was based on behavioral change theory and personalized to each subject. Texts were sent to the intervention arm 3 times/day with encouraging messages based on real-time activity data. Each subject had a personalized step count target which increased by 20% every 4 weeks. At the end of 12 weeks, the intervention arm took 1019 more steps per day than the control arm, an increase of 20% over baseline. We now hypothesize that increasing step counts will improve HRQOL in patients with PAH. We propose a randomized trial of smart texts versus usual care for 6 months. We will randomize 100 PAH patients to the mHealth intervention or usual care. All enrollment, monitoring, and data collection will occur remotely at Vanderbilt. We will enroll subjects across the United States, increasing generalizability. Our enrollment targets are feasible because we are supported by two large, existing PAH cohorts – the NIH-funded PVDOMICS Consortium and the Pulmonary Hypertension Association Registry. R61 Milestones (Year 1): IRB approval; establish DSMB; create REDCap database; Data Use Agreements; programming of text intervention; enrollment of first 10 subjects. In Aim 1 (primary endpoint), we will test the effect of a text-based mHealth intervention on HRQOL in PAH using the PAH-specific emPHasis-10 questionnaire. In Aim 2 (secondary endpoint), we will test the effect of an mHealth intervention on exercise capacity, measured by a highly feasible and reproducible supervised home-based 6MWD test. In an exploratory aim, we will examine the effect of the intervention on time to clinical worsening (composite of PAH therapy escalation, PAH hospitalization, and death) one year after randomization. This proposal will test a novel, highly scalable, and affordable mHealth intervention to improve clinically meaningful outcomes in patients with PAH.

NIH Research Projects · FY 2024 · 2022-09

Project Summary Mozambique has made progress towards their 95-95-95 goals, yet only 65% of people newly initiated in HIV treatment remain in care at 12 months. HIV treatment adherence has been undermined by a lack of compassionate health care service delivery. People living with HIV in our study region of Zambezia province report being treated disrespectfully by health care workers as one of the leading causes of treatment abandonment. Common complaints include insulting patients’ intellectual capacity, ignoring concerns with side effects or co-occurring infections, and refusing to treat patients seen as “unworthy”. While health care worker behavior is appalling, it is likely a reflection of high rates of burnout, job dissatisfaction, and frustration with patients they perceive to be uncooperative. The implementation of Estamos Juntos (We are Together) will allow us to test a multiprong intervention designed to address provider-barriers to delivering compassionate care via two synergistic components: (1) Resilience and well-being training for health care providers who have expressed low job satisfaction, frustration with delivering care in an extremely resource-limited setting, and burnout; and (2) Anti-stigma training for health care providers who see those with low socioeconomic status, low levels of education, and those living with HIV as “lesser-than” themselves. We propose to pilot test the implementation and impact of each psychosocial intervention individually, and in combination, using a randomized controlled trial design at four health facilities. We hypothesize that the facility where health care workers receive both resilience and anti-stigma training will see the greatest change in health care worker outcomes, including decreased stigmatizing attitudes, emotional exhaustion, and depersonalize of their patients, as well as increased resilience and job satisfaction. Patients receiving care and treatment from intervention providers will show increased retention and medication adherence, as well as improvements in health care services satisfaction, medical mistrust, and perceived stigma from health care providers. The Specific Aims of this study are to: (1) Evaluate the impact of resilience training only, anti-stigma training only, and resilience and anti-stigma training (vs. standard of care) on hypothesized mechanisms of behavior change among health care providers employing the RE-AIM framework through a cluster randomized controlled trial; and (2) Investigate the impact of provider training to reduce stigma and increase wellness and resilience on patient adherence to HIV treatment. Our study team has more than a decade experience developing and testing HIV and associated mental health interventions in Mozambique. If successful, our intervention can be tested through the R01 mechanism in a fully powered trial designed to improve both provider and patient health outcomes.

NIH Research Projects · FY 2025 · 2022-09

Project Summary/Abstract Puberty is among the most complex developmental transitions over the human lifetime. The timing (age of onset) and tempo (rate of change) of puberty can significantly and negatively impact physical, psychosocial and physiological functioning, especially in females. Autism spectrum disorder (ASD) is characterized by impaired reciprocal social communication and poor adaptation to change; thus, the onset and course of puberty marks a pivotal transition. In a series of cross-sectional studies, findings from the study team have shown: 1) advanced pubertal timing for female youth with ASD compared to typically developing females (i.e., breast development, menses) using rigorous assessment of pubertal staging; 2) distinct camouflaging behaviors in females with ASD when socially interacting with peers; 3) higher depressive symptoms in early adolescents with ASD, especially females; 4) sex-based differences in neural socioemotional processing in youth with ASD; and 5) elevated stress and arousal in youth with ASD compared to same-age TD peers that increases with age and pubertal development. Consistent across these studies is the unique profile of females with ASD and the potential deleterious impact of early puberty in this vulnerable, understudied population and developmental window. While the previous and ongoing pubertal development research is compelling, at the start of enrollment, 80% of the 10-year old females with ASD had already entered puberty compared to 29% of the TD group; therefore, the true onset of puberty could not be determined. Moreover, it was based on a relatively small sample of females with ASD 10-13 years (N = 35). Thus, a comprehensive exploration of pubertal maturation in a large sample of younger girls with ASD is needed. The overarching goal is to systematically examine the precise onset, tempo and course of pubertal, psychosocial and physiological development in females with ASD using a multimodal, multimethod, accelerated longitudinal design. A large sample of females with ASD (n=120) and TD (n=120) spanning 6 to 15 years will be followed over four years: Cohort 1 enrolled at 6-years (follow 6-9), Cohort 2 enrolled at 8-years (follow 8-11), Cohort 3 enrolled at 10-years (follow 10-13) and Cohort 4 enrolled at 12-years (follow 12-15). The aims will address three key areas. Aim 1: Pubertal development - will examine the timing, tempo and course of puberty based on physical development (Tanner stage (TS), body mass index (BMI), height, linear growth velocity (LGV)), hormones (Estradiol, Luteinizing Hormone (LH)), and menstrual cycle. Aim 2: Psychosocial profiles - will examine reciprocal social communication (CASS) and internalizing symptoms (anxiety and depression) at the onset and over the course of pubertal development. Aim 3. Physiological characterization - will simultaneously examine social functioning at the level of the central nervous system (EEG hyperscanning), HPA axis (cortisol) and peripheral nervous system (RSA) during naturalistic social interactions between same-sex peers. The findings will reveal the impact of the onset, tempo and course of puberty on females with ASD to improve education, inform services, and develop interventions, which are lacking.

NIH Research Projects · FY 2025 · 2022-09

Non-viral gene delivery systems are limited by their activity and targeted integration capability. Efficient and targeted integration of DNA into mammalian and human genomes remains a major challenge and its success would have wide impact for biotechnology and therapeutic applications. The piggyBac (PB) transposon system is the most active integrating non-viral gene delivery system and is a cut-and-paste DNA transposon that has been used for genome engineering of mammalian and human cells for more than 15 years. We have re-engineered the PB-transpososome (transposase with transposon DNA) based on the first- ever three-dimensional structure of the PB transpososome that we recently published with our collaborator Dr. Fred Dyda (Chen et al., Nature Communications, 2020). Our next-generation PB transpososome (ngPB) demonstrates greater activity and potential for targeted integration than was previously achievable. In specific aim 1, we will engineer and test ngPB for genome engineering of human cells. We will evaluate the integration site profile and copy number of transposon integrations per human cell. We will modify primary human T cells ex vivio and test their ability for cell therapy, and we will enable transposase protein transfection. In specific aim 2, we will engineer and test ngPB for gene delivery in vivo. We will evaluate gene delivery of reporter and therapeutic transgenes to mouse liver, test for efficiency in development of transgenic mice, and evaluate hybrid adeno-associated viral (AAV)-ngPB mediated gene delivery to difficult to reach organs. In specific aim 3, we will engineer and test ngPB for targeted integration in human cells. We will also map the protein-protein interaction domain of PB known to affect its target site selection in human cells and test PB protein modifications to allow greater flexibility in manipulating PB genomic target site selection. The proposed studies will be transformative for genome engineering and have broad impact for biotechnology and therapeutic applications.

NIH Research Projects · FY 2025 · 2022-09

Project Summary/Abstract There is a critical need for accurate, efficient, and portable substance use disorder (SUD) identification methods that support large-scale genetic studies in uncovering biological mechanisms helpful in SUD prevention and management. The long-term goal of this work is to support genetic discovery efforts that result in beneficial interventions for prevention and treatment of SUD. The overall objective in this proposal is to develop and evaluate an SUD phenotyping method that allows investigators to fine-tune phenotypes for their specific projects. Given most large-scale SUD phenotyping for genetics studies have relied on administrative billing codes (that undercount true cases) and binary outcome labels (that induce arbitrary dichotomization), the rationale for this project is that a system which includes a variety of data sources and generates a probabilistic outcome along a continuum is needed. The SUD phenotyping framework will support the inclusion of heterogenous electronic health record data types (including administrative billing codes, medication information, and unstructured text data) and will be evaluated in multiple organizations. Pairing these SUD phenotypes with genetic data will enhance our understanding of SUD mechanisms among individuals. That foundational work could ultimately result in the development of polygenic risk scores and clinical decision support systems that we could implement prospectively in clinical care. The proposed research is significant because researchers have a pressing need for SUD phenotyping approaches that can be customized to their research focus and available data. This proposal’s innovation lies in the creation of a “self-service” approach to SUD phenotype development in which a research team can specify their own phenotype definitions. The software will have a graphical user interface that makes the highest-yield rules/heuristics the easiest to use and can therefore be used by investigators with basic scientific programming knowledge. Through the activities outlined above, this innovation will directly accelerate genetic studies of SUD while simultaneously developing a precision phenotyping framework that can be applied to other disease domains.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY: Thyroid cancer is rapidly increasing in the U.S., with papillary thyroid carcinoma (PTC) as the most common sub- type. While most patients are cured of their thyroid cancer following initial treatment, a significant portion of patients develop aggressive disease. Some PTCs even develop into anaplastic thyroid carcinoma (ATC), a highly lethal and treatment-resistant disease with an abysmal 4-month survival. Cancer-associated fibroblasts (CAFs) have been shown to play an important role in the aggressive behavior and progression of some cancers, including pancreas and triple-negative breast cancer. Preliminary data from our group show that ATCs are defined by a large population of tumor-promoting CAFs that express Fibroblast Activation Protein Alpha (FAP) and Wnt-2 ligand. These CAFs sit along the tumor-stromal interface and likely support a de-differentiated and stem-like phenotype in adjacent tumor cells, as defined by tenascin-C (TNC) expression. We find that small numbers of these FAP+ CAFs are present in a subset of PTCs and serve as a robust biomarker of future disease progression. Our preliminary studies provide compelling evidence that CAF subsets and CAF-derived Wnt ligands drive tumor growth and progression, with elevated levels of Wnt signaling associated with poorly differentiated and aggressive disease. The overall goal of this proposal is to determine the role of CAFs in thyroid malignancy and how their paracrine Wnt signaling supports aggressive tumor behavior. To evaluate the role of CAFs in thyroid cancer, we have collected a cohort of ~300 patients with thyroid disease from Vanderbilt University Medical Center and the University of Washington. We have also created the only thyroid biobank with >50 primary-patient derived thyroid cancer organoids and corresponding CAF cultures. This biobank allows our team to study the thyroid cancer microenvironment across a heterogeneous landscape of morphologies and genetic alterations. Finally, we create a patient-derived xenograft mouse model for thyroid cancer to study the in vivo biology of thyroid cancer and to test new therapeutics (including anti-Wnt drugs) for thyroid cancers resistant to standard-of-care therapy. In this proposal, we will test the hypothesis that distinct CAF subsets (FAP+ Wnt-2+) are present in thyroid cancers and that their secretome drives Wnt- mediated paracrine signaling for tumor progression. In Aim 1, we will define the role of Wnt signaling in thyroid CAF heterogeneity in ATCs, PTCs, and composite tumors showing the transition between both morphologies. In Aim 2, we will define the molecular interaction between Wnt-2 ligand and Tenascin-C (TNC) to promote a stem-like phenotype in ATC. In Aim 3, we will investigate Wnt blockade for stromal reorganization and treatment of ATC. An understanding of the role of CAFs and Wnt signaling will dramatically improve the detection and treatment of the most aggressive forms of thyroid cancer. ATC has limited treatment options and a dismal average survival of ~4 months. Successful completion of these goals will lead directly to clinical trials of novel therapies for ATC that directly target the tumor-promoting stromal microenvironment.

NIH Research Projects · FY 2025 · 2022-08

Cardiovascular diseases are leading global causes of death and disability, presenting as interrelated phenotypes of atherosclerotic vascular disease, heart failure, and arrhythmias. They arise from interactions between environmental factors and common and rare genetic variants, including relatively common Mendelian lipid disorders, cardiomyopathies, and arrhythmias that collectively occur in at least 1/100 individuals. The availability of genetic sequencing is altering clinical management, but a major barrier to the widespread application of this practice is that the function of the vast majority of variants in key cardiovascular disease genes is unknown. Variant effect maps that define function for nearly all missense variants in a target sequence offer a way forward. This project brings together scientists at the forefront of variant effect mapping in diverse cellular systems, illuminating underlying cardiovascular biology, establishing relationships between variant function and human phenotypes, and working with others in multi-institutional collaborations. Our CardioVar team will generate a comprehensive atlas of variant effect maps for key cardiovascular disease genes. In Aim 1, we will develop, optimize, and validate a range of high-throughput cellular assays. We will use a range of generalizable (e.g. surface abundance) and bespoke (e.g. electrophysiological, lipoprotein uptake) assays to directly measure variant function in disease-relevant context. Assays will be assessed by their ability to discriminate pathogenic from benign variants. In Aim 2, we will use in situ targeted mutagenesis or insertion of variant constructs at a safe harbor site to generate pools of cells capturing all single-nucleotide changes in target genes. We will then deploy existing validated assays and those emerging from Aim 1 to generate and validate variant effect maps at scale. Functional scores and uncertainty estimates will be derived and evaluated, both by performance on pathogenic and benign variants and on correlation with discrete and quantitative phenotypes in clinical cohorts. In Aim 3, we will derive biological and clinical insights from variant effect maps. Discordant cases, where variant scores diverge from clinical annotation, will be further investigated in zebrafish, iPSC-cardiomyocytes, and automated patch clamping systems. Through a combination of hypothesis-driven analysis and machine learning models, we will reveal relationships among variant effects, protein structure, protein function, and human phenotypes. To optimize use of the atlas, we will provide a portal serving as a variant-centric decision support system for evaluating functional evidence of pathogenicity. We will release variant effect map data pre- publication via MaveDB (that we co-developed) and share all renewable variant assay reagents. The CardioVar atlas of missense variant effects, covering key cardiovascular disease genes, will be an essential and interpretable community resource for clinical and mechanistic studies of cardiovascular disease.

NIH Research Projects · FY 2025 · 2022-08

ABSTRACT Most patients with pulmonary arterial hypertension (PAH) die from right ventricular (RV) failure and all experience exercise limitation. No RV-specific therapies exist because the mechanisms underlying RV failure are poorly understood. The discovery of novel, potentially treatable causes of RV failure and exercise limitation would be an important advance in the treatment of PAH. RV steatosis may be a novel mechanism of RV failure in human PAH. We and others reported a high prevalence of insulin resistance in patients with PAH. We recently published that insulin resistance in PAH manifests primarily as abnormalities in lipid metabolism. Patients with PAH had elevated circulating free fatty acids and long-chain acylcarnitines and we found impaired mitochondrial fatty acid oxidation and RV lipid accumulation (steatosis) in a rodent model of PAH. We used proton magnetic resonance spectroscopy (MRS) to quantify RV lipid in vivo and showed that RV lipid content is over 10-fold higher in humans with PAH compared with matched controls and may be modifiable with metformin. In autopsy RV specimens, we found increased ceramide, a mediator of lipotoxicity, and identified candidate plasma surrogates for RV steatosis. Finally, we present new evidence of skeletal muscle steatosis in humans to demonstrate that insulin resistance is a systemic feature of PAH. The clinical relevance of RV steatosis in human PAH is unknown. RV steatosis is common in both heritable and idiopathic PAH and is not an end-stage phenomenon, which suggests potential for therapeutic intervention. We hypothesize that abnormal lipid metabolism in PAH leads to delivery of fatty acids in excess of RV oxidative capacity, resulting in steatosis and lipotoxicity. Our objectives are to: 1) Define the relationships between RV steatosis, RV function, and exercise capacity; 2) Identify mechanistic drivers of RV steatosis including BMPR2 expression and lipid metabolism; 3) Examine lipid metabolism in PAH skeletal muscle as a potential driver of reduced functional capacity; and 4) test the response of RV and skeletal muscle steatosis to metformin. In Aim 1 (clinical relevance) we will measure RV and LV lipid in participants with heritable, idiopathic, and scleroderma- associated PAH. Participants will undergo the 6-minute walk test, cardiopulmonary exercise testing, and will be followed for clinical events. A subgroup will undergo repeat MRS at four timepoints over three years to determine the natural history of steatosis. We will measure RV lipid in participants before and after metformin therapy in a separate on-going study. In Aim 2 (mechanism), we will perform metabolomic/lipidomic profiling of peripheral and coronary sinus plasma and measure BMPR2 expression to identify potential drivers of steatosis. In Aim 3 (specificity), we will perform MRS on skeletal muscle in Aim 1 participants and matched healthy controls to clarify the systemic effects of lipid metabolic defects in PAH. We will also test the effect of metformin on skeletal muscle lipid content and fatigue. The proposed studies address an important knowledge gap in PAH pathophysiology by interrogating a newly discovered, novel mechanism of RV failure in humans.

NIH Research Projects · FY 2025 · 2022-08

PROJECT SUMMARY/ABSTRACT Juvenile idiopathic arthritis (JIA) is an autoimmune arthritis in children characterized by chronic joint inflammation. T helper type 1 (Th1), Th17, and pathologic Th17.1 cells and the inflammatory cytokines produced by these cells, interferon gamma (IFNγ) and interleukin-17 (IL-17), are implicated in JIA pathogenesis. A major knowledge gap is to understand how these inflammatory Th cells and cytokines develop. This proposal outlines a research and training plan focused on studying how IL-17 and dual IFNγ-IL- 17 producing cells inappropriately develop during JIA Th1 differentiation. A goal for these studies is to identify biologic factors that can be used to improve diagnostic and therapeutic decisions. The investigator conducted studies that described: 1) polyarticular JIA circulating cells that underwent Th1 differentiation produced high levels of IL-17 and IFNγ and dual IFNγ-IL-17 producing cells, 2) a JIA patient with a rare loss-of-function GATA3 mutation exhibited an exaggerated form of this phenotype, and 3) additional JIA patients carry rare protein-coding mutations in genes important for Th1 differentiation. The proposal’s central hypothesis is that JIA Th1 differentiation inappropriately produces IL-17 and Th1.17 cells and rare genetic mutations contribute to this phenotype. The proposed Specific Aims test this hypothesis. Aim 1 identifies the role of novel genetic mutations from JIA patients in production of IL-17, IFNγ, and Th1.17 cells during Th1 differentiation. Aim 2 identifies the cytokine and STAT signaling pathways that lead to the production of IL-17 and Th1.17 cells during polyarticular JIA Th1 differentiation. This proposal involves translational studies in human cells using advanced cytometry, molecular biology, and next generation sequencing. A major focus of this proposal is to support Dr. Patrick’s development as a physician-scientist. Her career goal is to study the pathogenesis of JIA in a basic research program and identify biologic factors that generate novel therapeutic targets and improve diagnostic and therapeutic decisions. She will accomplish this goal through career aims to become an immunology expert, gain advanced expertise in immunologic techniques, establish proficiency in the use and analysis of next-generation sequencing, and acquire skillsets to become an independent principal investigator. The research environment for the proposal is outstanding. Dr. Patrick has full departmental and institutional support for the development of her research program. Her mentors are immunologists with expertise in the planned advanced techniques. Vanderbilt has excellent facilities and shared resources for training in these techniques. Her mentoring team includes experts in immunology, gene regulation, and rheumatologic disease to guide development of her independent research program. This K08 will support the generation of data and lead to publications providing insight into the pathogenesis and genetics of JIA and in support of a successful R01 application.

NIH Research Projects · FY 2024 · 2022-08

Abstract Great variation exists in opioid consumption after similar surgeries, with some patients using few opioids while others consume opioids well beyond the expected acute pain episode. Prolonged opioid use is considered one of the most common surgical complications. Recent models developed to predict the variability in postoperative opioid use focus primarily on clinical and psychological survey data, but leave a significant proportion of the variability unexplained. As it is unclear whether prolonged opioid use results from prolonged pain or other factors, this project proposes to extend the previous research by prospectively examining important pain sensitivity and biomarker predictors of pain to determine their effect on opioid use. Pain is a biopsychosocial phenomenon, so the inclusion of these important biological variables will lead to increased mechanistic understanding of postoperative opioid consumption variability and enable better management of postoperative pain by providing potential new therapeutic targets. One such understudied variable is an individual’s underlying pain sensitivity, measurable by quantitative sensory testing (QST). Use of QST to predict inpatient opioid use and pain has led to inconsistent findings, but recent studies highlight a promising role for QST to predict postoperative home opioid use. Speculation has long focused on genetic influences as a factor in variability in pain sensitivity and opioid use. Candidate gene and whole genome-wide studies have proven to be inadequate to study the complex trait of pain, resulting in small effect sizes and lack of reproducibility. Assessing gene expression is an approach that can be used to augment the research on postoperative pain. Gene expression analyses in blood have identified changes that confer vulnerability for the transition from acute to chronic low back pain, lending support to the hypothesis that changes in gene expression from preoperative to early and later postoperative time periods may identify early markers for the transition to prolonged postoperative pain. By integrating genetic and gene expression data, we will assess whether acute surgical pain leaves signatures in the blood that can be identified through gene expression which can influence postoperative pain sensitivity and subsequent opioid use. Our proposal focuses on the total knee arthroplasty (TKA) population, as this population demonstrates one of the highest variabilities in postoperative pain and opioid use, and is projected to increase 700% within the near future. This project proposes to innovatively incorporate standard demographic, clinical, and psychological variables with novel neurophysiological, genetic, and gene expression data to holistically examine the variability in postoperative pain, and ultimately opioid use through the biopsychosocial lens.

NIH Research Projects · FY 2026 · 2022-08

Articulatory impairments are common in talkers with dysarthria and have been shown to contribute most to speech intelligibility loss. However, the treatment of articulatory impairments remains challenging; primarily, because knowledge about articulatory impairment patterns and how they vary across talkers is limited. The Mayo Clinic dysarthria classification system links neurological conditions to specific, auditory-perceptually defined dysarthria types with presumably distinct motor impairment patterns. Based on this framework, prominent textbooks have recommended to specifically target the motor impairments that presumably underlie each dysarthria type. Such an intervention approach, however, still lacks empirical support and is difficult to apply to mixed dysarthria types. Based on the rationale that virtually all talkers with dysarthria exhibit imprecise articulation, impairment-nonspecific behavioral treatments (e.g., loud, clear, slow speech) have gained popularity and are commonly used as therapeutic interventions for talkers with mild to moderate dysarthria regardless of their underlying disease. This rationale, however, ignores the fact that a variety of articulatory behaviors can yield the same auditory-perceptual consequences. Indeed, highly heterogeneous articulatory performance patterns have been reported across and within disease types and motivated us to test the feasibility of a new, taxonomic (data-driven) approach to dysarthria classification based on speech kinematic measures. Our pilot work on a cohort of 28 talkers with Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), and multiple sclerosis (MS) revealed six dysarthria subgroups with unique articulatory impairment profiles addressing temporal and spatial characteristics of vocal tract adjustments as well as labial coupling. Three disease-dominant subgroups and well as three mixed-disease subgroups were identified. This proposed project seeks to expand upon these preliminary findings. In Specific Aim 1, we will classify the articulatory performance profiles of 160 talkers with varying underlying disease types [PD, ALS, MS, Huntington's disease (HD)]. To allow for a clinical interpretation of the kinematic findings, articulatory performance of talkers with dysarthria will be referenced to age- and sex-specific control groups. We will also determine which kinematic measures differentiate talkers with dysarthria and if disease-type varies systematically across articulation- based dysarthria subgroups. To ensure a rapid translation of our kinematic-based classification approach into clinical practice we will determine how perceptual-based clinical ratings of articulatory performance map onto findings of kinematic measures (Specific Aim 2). Raters will judge the articulatory performance of the same 160 talkers with dysarthria using auditory- and visual-perceptual ratings scales. Study outcomes will advance the field's understanding of articulatory impairment patterns and how they vary in dysarthria. Findings are critical to elucidate subgroup-specific articulatory mechanisms of intelligibility loss and recovery. Ultimately, this work will facilitate personalized dysarthria management and the development of new, impairment-specific interventions.

NIH Research Projects · FY 2025 · 2022-08

Project Summary/Abstract This proposal is to develop a pre-shimmed parallel transmit array, an optimized receive array, and an RF/ΔB0 array to correct the severe B1 inhomogeneity, maximize the signal-to-noise ratio (SNR), and correct B0 inhomogeneity in simultaneous human brain and spinal cord MR imaging 7 Tesla (T). Simultaneous functional imaging of the brain and spinal cord can provide valuable insight into interactions and processing pathways between these organs in normal and abnormal states of spinal cord injury, chronic pain, and motor disease. It is emerging as a new tool to study the central nervous system and is necessary to enable new investigations of task-based and resting-state sensory/motor processing throughout the cerebrum and spinal cord and shed new light on the nature of resting-state networks within the cerebellum and spinal cord. 7T MRI offers new opportunities to visualize structures of interest with high spatial resolution and enhanced conspicuity and to detect brain function and networks with greater sensitivity. However, at high fields, B1 and B0 inhomogeneities, and the lack of optimized receive coils for some specific applications are major challenges that limit imaging performance. Existed designs are aimed at either brain-only or spinal-cord-only applications, and none have solved all the challenges mentioned above. Moreover, the performance of these designs is limited by the small number of transmit channels available from scanner vendors, and a lack of optimization for actual imaging applications. The first goal of this project is to build a pre-shimmed transmit array which compresses 48 basic coils into 8-“virtual” coils with RF pulse jointly optimized weights, to maximize the transmit performance of standard 8-transmit-channel 7 Tesla scanners. The second goal of this project is to build a close-fitting massive- element receive array with optimum coil geometry/layout/size, to provide high SNR and excellent parallel imaging performance in both the whole brain and the spinal cord. The third goal of this project is to build routing-optimized low-profile RF/ΔB0 array to correct B0 inhomogeneity with less hardware complications. The optimization algorithms, electromagnetic simulation models, and electric/mechanical designs of the final pre-shimmed transmit array, high dense receive array and the routing-optimized ΔB0 arrays, will be distributed for open access. These transmit, receive, and ΔB0 arrays do not depend on the vendors’ platform and can be easily transferred to other 7T sites, with benefits for the entire community.

NIH Research Projects · FY 2025 · 2022-08

PROJECT SUMMARY/ABSTRACT The treatment of patients with Huntington disease (HD) is quickly entering a new era, with gene- silencing and disease-modifying therapies now in clinical trials. While current studies include patients with motor manifest disease, future clinical trials will require interventions in presymptomatic HD gene carriers in the hope of altering the course of the disease prior to the onset of motor symptoms. However, the critical question remains how to target potential therapies and measure outcomes in the large and heterogenous population of youth and young adults at risk for HD. Recent longitudinal studies have shown that cognitive and behavioral changes emerge decades before motor symptom onset, but the full spectrum of these symptoms has not yet been well-defined, and precisely which symptoms occur first and how to measure them remain matters of debate in the field. Furthermore, data on individuals under age 18 are largely lacking. Our preliminary data demonstrate that children who are at risk for HD face a multitude of challenges, including executive dysfunction, chronic stress, impaired coping skills, and significantly elevated neuropsychiatric symptoms, including depression, anxiety, and impulsive behaviors. However, the neurobiological basis of these symptoms, social and environmental contributing factors, and the potential impacts of mutant huntingtin protein and aberrant neurodevelopment remain unknown. The aims of this career development award are (1) to investigate the association between CAG repeat expansion, adverse childhood experiences, and externalizing risk-taking behavior in youth at risk for HD; (2) to examine alterations in response inhibition that may underlie impulsive behaviors in this cohort; and (3) to identify neurophysiological markers of inhibitory control that may represent modifiable treatment targets for future therapeutic trials. This proposal is supported by a multidisciplinary team of mentors with expertise in neuropsychiatric manifestations of neurodegenerative disorders, child psychology and human development, and electrophysiology. My overarching goal is to become an independent physician-scientist with unique expertise in the assessment of prodromal behavioral manifestations as early markers of neurodegenerative conditions. This proposal will build on my previous experience and will provide a unique training opportunity to develop new skills in neurophysiology, longitudinal data analysis, and the application of current neurodevelopmental models of psychological disorders that will allow me to conduct future independent investigations examining the developmental course of aberrant behaviors and impulse control in a prospective, longitudinal cohort from youth to adulthood. This work will fill a critical gap in our knowledge regarding the earliest manifestations of HD and will help to better target potential treatments during the premanifest phase of the disease.