University Of Pennsylvania

NIH Research Projects · FY 2025 · 2022-07

Covid-19 exposed differences in healthcare access and health outcomes (health opportunities) across the population—but in an unprecedented turn, policymakers deployed Disadvantage indices (DIs), which were novel tools, to address these differences within, and outside of the pandemic. Rapidly and widely adopting a proposal by the National Academies of Science, Engineering and Medicine (NASEM), a majority of US states (n=34) used DIs in vaccine allocation plans. DIs are place-based statistical measures of advantage and disadvantage that integrate Census data such as income, education, or quality of housing, to rank geographic areas as small as neighborhoods. Due to severe scarcity, DIs were used to increase vaccine shares for disadvantaged areas. This use mitigated the risk that groups, who typically had worse health before the pandemic, would incur a further burden during vaccine allocation. At the same time, the rapid adoption and wide range of DI uses leaves unclear what the optimal uses of DIs are within and outside of health emergencies. Our goal for this study is to determine the strengths and weaknesses of using DIs for the Covid-19 pandemic, future pandemics, public health, and clinical care. As a highly interdisciplinary team collaborating with a community advisory board, we propose an observational study with 2 aims. First, we will identify the impact, strengths, and weaknesses of using DIs in Covid-19 vaccine allocation. We will evaluate the impact of the 3 most frequently used DIs on Covid-19 hospitalizations, deaths, and vaccination rates, using predictive modeling and analyses of states' actual vaccine-roll-out. We will also conduct qualitative interviews of key stakeholders to identify facilitators and barriers to using DIs with vaccine allocation. Second, we will identify the possible strengths and weaknesses of using a wider range of DIs in public health and clinical care outside of emergency settings, including for health-related social needs. From the fourth year of the study, we focused on analyzing the intersection of DIs and central health outcomes and opportunities, such as life expectancy, cancer rates, and access to basic needs such as electrical or other utilities. We will use interviews and other methods to determine how key stakeholders, including public health, hospital, and research leaders, have used and rank concrete uses of DIs, identified from the literature. We complement expert views with two innovative nationally representative survey-experiments, and engaging communities in group deliberations.

NIH Research Projects · FY 2025 · 2022-07

PROJECT SUMMARY Staphylococcus aureus is a bacterial species that causes infections of the skin, lungs, blood and internal organs. These infections can have their onset in the community or in the health care setting, including those caused by antimicrobial-resistant strains called methicillin-resistant S. aureus (MRSA). About 28-50% of all people are asymptomatic carriers of this species on their bodies. The interaction of S. aureus strains in the community and in hospitals has not been adequately studied. While we think that S. aureus can spread from person to person in hospitals, we are not sure how often that happens, if some strains spread more readily than others in the hospital, and how often infections result from the S. aureus bacteria that patients bring with them on their own bodies. Also, it is unclear how important fomites (inanimate objects) or the bodies of healthcare workers (HCWs) are as intermediate carriers in spreading S. aureus from patient to patient in the hospital. This study will be based on data from whole genome sequencing (WGS) of S. aureus collected in two units at the Hospital of the University of Pennsylvania (HUP). For 24 months, we will test all patients upon admission for S. aureus colonization of 4 body sites. For a subset of subjects colonized with S. aureus, we will test fomites in their hospital room for S. aureus. We will choose control patients who are not colonized with S. aureus on admission and test their room fomites to determine what the risk factors are for acquisition and spread of S. aureus in the hospital. Using WGS we will test all of the bacteria causing infections on the study units to see how often specific strains spread, and we will determine whether some genetic traits of S. aureus make them more likely to spread and/or more likely to cause infections. In addition, each month, we will test a sample of the noses and gloves of HCWs on study units to see if they carry S. aureus. We will test common areas of these units to see if fomites carry S. aureus. We will determine if any of the spreading S. aureus strains are found on HCWs or common-area fomites. We will examine all infecting S. aureus isolate genomes from the entire 850-bed HUP during an overlapping 2-year period to identify evidence for distant spread of S. aureus causing an infection. We hope to better understand the genetic markers of virulence and transmissibility in S. aureus. We will test the hypothesis that strains of S. aureus that spread develop a distinct collection of plasmids, which are mobile genetic elements coding for antibiotic resistance or virulence factors that S. aureus may or may not carry. With these results, we may be able to identify patients carrying high-risk S. aureus isolates and develop strategies to prevent the spread of this bacterium in the future.

NIH Research Projects · FY 2026 · 2022-07

Tendons can withstand large forces due to a highly aligned, dense collagen matrix. However, their low cellularity and relative inability to recruit reparative cells post-injury, as well as susceptibility to excessive scarring results in loss of tendon structure and mechanical function. Type I collagen (Col1) is the primary collagen of healthy tendon and type III collagen (Col3) is a minor constituent that increases in response to injury. In other Col1-rich tissues such as skin and bone, Col3 directs reparative cell activities by regulating early cellular infiltration to promote healing, as well as collagen deposition, architecture and crosslink formation, supporting an early critical role of Col3 in wound healing, which has not been studied in tendon. Adding to the importance of understanding a role for Col3 in tendon, Col3 levels in aging tissues are reduced and in aged tendon, we have shown inferior healing which raises the possibility that age-induced Col3 loss increases susceptibility to poor tendon healing in aging populations. While Col3 may orchestrate cellular activities and fate that are critical for an optimal reparative response post-injury at early stages in tendon, its persistent expression in the remodeling phase may compromise the desired healing response. Therefore, our overall objective is to delineate mechanisms by which the temporal expression of Col3 modulates the injury response throughout tendon healing, as well as its differential effect throughout aging. Specifically, we will test the hypothesis that Col3 is crucial for early tendon healing, but that its continued expression during remodeling is detrimental. To test this, we generated a novel inducible Col3 deficient (i.e., Col3a1F/+, Col3a1F/F) mouse model to determine the dose-dependent effects of Col3 by temporal targeting of Col3. Using this approach, we will define the regulatory roles of Col3 throughout tendon healing at the time of injury, during the early proliferative phase, and during remodeling. In addition, by knockdown of Col3 in young, middle-aged, and old animals, we will evaluate the effects of aging on tendon healing with altered Col3. Understanding the role of Col3 throughout healing will lead to clinically relevant insights to improve outcomes following tendon injury in patients of varying ages. The study aims are: Aim 1: To define the age-sensitive mechanistic role(s) of Col3 throughout tendon healing by knocking down Col3 at the time of tendon injury. Aim 2: To elucidate the age- sensitive mechanistic role(s) of Col3 in directing reparative cell activities and prolonged effects of knocking down Col3 during the proliferative phase of tendon healing. Aim 3: To define the age-sensitive mechanistic role(s) of Col3 in tendon repair by knocking down Col3 during the remodeling phase of tendon healing. This study will define the critical temporal roles of Col3 in response to tendon injury throughout aging. We will utilize a novel mouse model coupled with rigorous hierarchical structure/function assays to define the role of Col3 in directing interplay between cellular activities and matrix assembly/organization in tendon healing.

NIH Research Projects · FY 2024 · 2022-07

Over 100 million Americans suffer from chronic pain, experiencing persistent nociceptive sensory and negative affective symptoms. Currently available pain treatments are inadequate at safely and effectively relieving both the sensory and affective features of chronic pain, partly because of their lack of specificity to modulate distinct neural cell-types and processes. While diagnostic imaging tools have correlated the affective unpleasantness of chronic pain with dysfunction across broad brain regions-such as the basolateral amygdala (BLA) and nucleus accumbens (NAc)-they lack specificity to identify individual functional cell types that might underlie this maladaptive plasticity. A key first step towards identifying and targeting affective nociceptive neural circuits is to visualize the dynamics of nociceptive transmissions between affective-motivational brain regions during the transition from acute to chronic pain. The research goal of the proposed project is to functionally characterize a nociceptive BLA to NAc circuit that contributes to negative affective-motivational behavior in acute and chronic nociceptive states. Dysfunction of the BLA has been implicated in chronic pain and neuropsychiatric disorders through the process of assigning valence to internal and external sensory information. The BLA contains a functional subpopulation of negative valence neurons essential for the emotionally aversive aspect of nociception (BLA"0 ci). Inhibition of BLAnoci neurons reduces affective-motivational responses to noxious stimuli in acute and chronic nociceptive states. BLA neurons send long-range axons to many downstream targets, including the NAc, a striatal structure important for driving appetitive and aversive behaviors. Preliminary data suggests that BLAnoci neurons project to the "limbic" NAc Shell subregion (NAcSh). Furthermore, there is a group of neurons highly responsive to noxious stimuli within the NAcSh (the "inner-horn"; NAcSh1H) that a bundle of BLAnoci axons strongly innervate. However, it is unknown if BLAnoci neurons transmit affective nociceptive information to the NAcSh1H nor if NAcSh1H neurons are essential for driving nociception-related negative affective-motivational behaviors. Aim 1 will functionally characterize the BLAnoci to NAcSh1H circuit in acute and chronic nociceptive states using in vivo calcium imaging and optogenetic activation of BLAnoci axon terminals in awake behaving subjects. Aim 2 will image and manipulate NAcSh1H neuron activity in acute and chronic nociceptive states using calcium imaging and optogenetic manipulation of NAcSh1H neural activity. Completion of these proposed aims will result in the characterization of a potentially key affective circuit between the BLA and NAcSh1H encoding acute and chronic nociceptive information. The outcome of this proposal will help the field of affective pain neuroscience better understand neural circuit function in healthy and pathological states. Furthermore, this research may inform future translational investigations into treatments for the negative affective symptoms of chronic pain. Completion of this fellowship will achieve the training goals of expanding the technical expertise of Ms. Wojick in systems neuroscience and facilitate her career goal of becoming a leading expert in affective pain research.

NIH Research Projects · FY 2025 · 2022-07

Ribosome RNA (rRNA) modifying enzymes are important factors in the assembly and function of ribosomes, dysregulation of which is frequently seen in human hematopoietic disorders. We only have limited knowledge of the catalytic role of rRNA modifying enzymes in the physiological and pathological processes of human hematopoiesis, although a few studies characterized the importance of rRNA modifications in these processes. Preliminary data (CRISPR screening) from my laboratory shows that the ablation of DIMT1 (an 18S rRNA N26,6A-dimethylation (N26,6A) methyltransferase) significantly impairs cell viability of hematopoietic stem progenitor cells (HSPCs). DIMT1 is highly expressed in HSPC, and its expression decreases in the differentiated lineages. However, the molecular function and detailed mechanism of DIMT1 in human hematopoiesis remain elusive. Even though DIMT1-mediated rRNA modifications are highly conserved, the catalytic activity of DIMT1 is dispensable for proper ribosome biogenesis. However, our recent results suggest that the catalytic activity of DIMT1 is required for the viability of HSPCs, while the catalytic-independent role of DIMT1 is important for 18S rRNA processing and ribosome biogenesis. This implies that DIMT1 may have a necessary role as a scaffold in ribosome biogenesis separate from its catalytic activity. Furthermore, we show that the catalytic role of DIMT1 is indispensable for the expression of genes involved in the Fanconi anemia and other DNA repair pathways. However, whether and how DIMT1-mediated N26,6A installation in 18S rRNA influences cell viability and DNA repair pathways in HSPCs are not known. Here, our goal is to understand the molecular function and the mechanistic details of the catalytic role of DIMT1 in the regulation of cell viability and gene expression in HSPCs. Specifically, Aim 1 will investigate the catalytic role of DIMT1 as an 18S rRNA methyltransferase in HSPC cell viability and protein synthesis. Aim 2 will investigate the molecular mechanism by which DIMT1- mediated N26,6A in 18S rRNA controls translation elongation using an in vitro eukaryotic translation system. Aim 3 will investigate the structure-function relationships and catalytic mechanism of DIMT1. Results from these studies will provide novel insights into how the catalytic role of DIMT1 impacts HSPC cell viability and the expression of genes Fanconi anemia and other DNA repair pathways. The understanding we will gain can be broadly applicable to hematopoietic studies and many other biological disciplines.

NIH Research Projects · FY 2026 · 2022-07

ABSTRACT Atopic dermatitis (AD) is a common, chronic inflammatory disease of the skin that affects 10% of adults. AD is associated with major health-related quality-of-life (QoL) and psychosocial impairments that exceed those of other serious chronic diseases such as heart disease, diabetes, and hypertension. Despite the major physical and emotional burdens that accompany AD, it remains an understudied skin disease among adults because, until recently, AD had been incorrectly considered a childhood disease that remits in adulthood. Importantly, our preliminary data reveal racial/ethnic differences in the severity of, QoL impact of, and health care utilization for AD that suggest the presence of disparities in AD outcomes that deserve further study. Considering the anticipated diversification of the U.S. population, such disparities are only expected to worsen if left unaddressed. However, a critical barrier to identifying, understanding, and ultimately eliminating racial/ethnic disparities in AD outcomes among adults exists due to the lack of adult AD cohorts with sufficient racial/ethnic diversity and individual-level social and environmental contextual data that are necessary to comprehensively evaluate the causes of such disparities. In order to address this barrier, we propose to create a cohort of racially and ethnically diverse adults with AD within the socioeconomically disadvantaged and medically underserved, urban neighborhood of North Philadelphia, Pennsylvania. North Philadelphia adult residents represent a population that is vulnerable to a large burden of AD. With this cohort, we aim to perform a longitudinal cohort study to: i) confirm and further characterize existing racial/ethnic disparities in AD outcomes and health care utilization for AD among adults, and ii) identify the individual behavioral, social, and environmental contextual factors that simultaneously contribute to racial/ethnic disparities in AD outcomes. We will address these aims utilizing a novel transdisciplinary approach and a sequential mixed methods study design in order to incorporate the lived experience of adults with AD in our studies. We hypothesize that specific and potentially modifiable behavioral, social, and environmental factors contribute to racial/ethnic disparities in AD outcomes. The new knowledge that will be gained from the proposed work is essential to informing future individual- and community-level interventions to reduce racial/ethnic disparities and improve AD outcomes among similar urban adult populations.

NIH Research Projects · FY 2024 · 2022-07

Pathogenic variants in the gene SCN3A, which encodes the voltage-gated sodium (Na+) channel subunit Nav1 .3, cause SCN3A-related neurodevelopmental disorder (SCN3A-NDD), a recently identified condition defined by treatment-resistant epilepsy, severe to profound intellectual disability (ID), and, surprisingly, malformation of cortical development (MCD; abnormal structural development of the cerebral cortex). There is no known role for Na+ channels in structural brain development nor any basis for brain malformation caused by Na+ channel variants, and it remains unclear how genetic variants in SCN3A lead to MCD, epilepsy, and/or ID. This proposal seeks to define the mechanistic underpinnings of this poorly understood disorder in order to define the normal physiological role of Nav1 .3 and to inform therapeutic discovery or preventative measures for SCN3A-NDD, a devastating disease which currently has no treatment or cure. The prominence of MCD among our uniquely large patient cohort combined with known high embryonic expression of SCN3A in the brain motivates my central hypothesis that pathogenic variants in SCN3A cause epilepsy and MCD in SCN3A-NDD by producing pathological Na+ currents in developing cortical neurons leading to perturbed excitability and altered neuronal migration. Proposed experiments will functionally assess specific patient variants in SCN3A in our cohort and establish the relationship between individual genetic variant, channel dysfunction, neuronal excitability, structural development, and clinical presentation. Aim 1 will ascertain the link between genetic variation in SCN3A and Na+ channel dysfunction by determining the biophysical properties of wild-type versus variant Nav1 .3 in a transiently transfected (HEK-293T) cell system. Aim 2 will assess how altered channel activity impacts neuronal electrical excitability, anatomy, and maturation via generation of human excitatory cortical neurons derived from control or SCN3A variant-expressing induced pluripotent stem cell (iPSC) lines. A novel Nav1 .3-selective compound PF-6651385 will be employed to determine potential to pharmacologically correct dysfunction at the level of the channel (Aim 1) and/or neuron (Aim 2). This proposal using advanced model systems to interrogate pathogenic mechanisms of SCN3A-NDD will advance my training towards a career in translational neuroscience while providing novel insight into a potentially fundamental role for a Na+ channel in brain development and elucidating pathomechanisms underlying SCN3A-NDD to progress towards targeted therapies for patients suffering from this disorder.

NIH Research Projects · FY 2025 · 2022-06

PROJECT SUMMARY Group 1, 2, and 3 innate lymphoid cells (ILC1, ILC2, and ILC3) are immune effector cells that contribute to tissue homeostasis and host defense against nearly all classes of pathogens, but their dysregulation also play key roles in prevalent diseases such as cancer, obesity, asthma, and colitis. The transcription factor (TF) networks that control the development and functions of the different groups of ILC have recently been identified. Yet how the chromatin accessibility landscape and the 3-dimensional (3D) genome architecture determine the development, homeostasis, and effector functions of ILC is largely unknown. Thus, the overarching goal of this proposal is to uncover how the 3D genomic and epigenetic architecture regulate the development of each ILC subset and to the development of allergic airway inflammation. It is now well-stablished that the transcriptional repressor Id2 determines the commitment and identity of the ILC lineage. As such, Id2 expression is now considered a hallmark of all ILC subsets in mice and humans. Our preliminary data indicates that Id2 expression is controlled in ILC1, but not ILC2 or ILC3, by specific long-range DNA interacting loops between specific distal cis-regulatory elements (cis-RE) and the Id2 promoter. Moreover, we showed that ablation of these promoter- cis-RE interactions in mice leads to a dramatic reduction in ILC1 in multiple tissues, while the development and functions of ILC2 and ILC3 were unaltered. Thus, our findings indicate for the first time that Id2 expression is regulated by long-range DNA interacting loops between the Id2 promoter and distal cis-RE in an ILC-subset specific manner. Moreover, it indicates that ablating these cis-RE is a powerful strategy to generate genetic tools to study the roles of each ILC subset in the context of an otherwise intact immune system. Yet how the chromatin accessibility landscape and the 3D genomic architecture determines Id2 expression specifically in ILC2 and ILC3 remains unknown. Thus, in aims 1 and 2 of this project, we will use novel genetic tools that we generated, single cell sequencing technologies, and HiC to elucidate how chromatin folding and accessibility determine the development and functions of ILC2 and ILC3 through the regulation of Id2 expression. In aim 3, we will exploit the specificity of these regulatory mechanisms to study the functions of ILC2 during allergic airway inflammation in the context of an otherwise intact immune system. Collectively, these studies will answer the long-standing question of how Id2 expression is controlled to drive the ILC fate. Moreover, it will generate an atlas of the 3D genomic landscape of each ILC subset, which that will allow us to identify unknown non-coding regulatory regions are critical for the function and development ILC1, ILC2, and ILC3.Importantly, through the identification of specific regulatory mechanisms in each ILC subset, we have created novel mouse genetic tools to study the functions of each group of ILC in the context of an otherwise intact immune system, which might unveil novel therapeutic approaches to target ILC during inflammatory disorders.

NIH Research Projects · FY 2025 · 2022-06

Project Summary/Abstract Non-small cell lung cancer (NSCLC) is a devastating disease with a poor outcome. When a patient is first diagnosed with NSCLC, the person would typically undergo diagnostic bronchoscopy and/or therapeutic surgery. These two procedures have technical challenges that limit their success such as inaccurate biopsies, failure to locate nodules and lymph nodes, missed occult tumors, and positive margins. As a consequence, there is a 40% failure rate for these procedures. This Project will use optical imaging in the near-infrared (NIR) range to generate practical solutions to these problems and make these two common procedures safer, more efficient, and have improved outcomes. The goal of this Project is to test the hypothesis that intraoperative imaging in the NIR spectrum with targeted molecular tracers can identify tumor cells in order to improve procedures that are utilized for the management of patients with NSCLC. The innovation of this Program Project will be in the matrix of cocktail dye development, new NIR camera devices, and clinical translation. There are three specific goals of this Project. First, to develop a set of targeted near-infrared molecular imaging contrast agents that are sensitive and specific to a range of distinct targets on NSCLC. Second, to develop and integrate highly NIR spectral sensitive imaging platforms into commonly used white light imaging devices so they can be utilized during clinical procedures. And third, to apply these innovations to solve common clinical problems in order to impact patient care. If successful, the molecular optical imaging solution presented in this proposal would be immediately applicable to several hundred thousand lung cancer patients each year.

NIH Research Projects · FY 2026 · 2022-06

PROJECT SUMMARY/ABSTRACT: The goal of this K23 Mentored Patient-Oriented Research Career Development Award is to support the applicant in developing the critical skills necessary to become an expert in increasing access to evidence-based treatment for opioid use disorder (OUD) at touchpoints for care that individuals with OUD are likely to frequent. The proposed research focuses on NIDA's priority area of addressing real-world complexities by developing implementation strategies to improve treatment continuity after hospitalization in patients with OUD. Hospitalizations related to OUD and its complications have increased dramatically over the last few decades, leading to high morbidity, mortality, and healthcare costs. Hospitals are increasingly recognized as critical touchpoints for engaging individuals with OUD in treatment, and randomized controlled trials support the efficacy of initiating medications for OUD (MOUD) in acute care settings to increase treatment engagement following discharge. However, hospital-initiated treatment is insufficient if patients do not continue care in the community. Despite the robust literature on care transitions after hospital discharge for patients with other chronic conditions, there is limited evidence about the best strategies to optimize care engagement for patients with OUD after hospitalization. We need effective, scalable strategies for linking patients with OUD from acute care to ongoing treatment, including continuation of MOUDs. The research objective of this proposal is to develop and pilot test strategies to facilitate transitions of care for patients with OUD from the hospital to the community. The specific aims are: (1) Use a mixed-methods approach to identify determinants of effective care transitions for patients with OUD from acute care to outpatient treatment at the patient, provider, and system level, (2) Partner with hospital and community stakeholders and use implementation mapping to develop a multicomponent, modular toolkit to facilitate transitions from acute care to community addressing multi- level barriers, and (3) Conduct a pilot trial of the care transition toolkit. The primary outcome will be treatment engagement at 30 days, and secondary outcomes will include MOUD and substance use, service use, feasibility, acceptability and patient satisfaction. The mentorship team brings together experts in health services research, implementation science, and care delivery interventions for OUD as well as stakeholder advisors from hospital and community settings. This Mentored Research Scientist Development Award builds on Dr. Lowenstein's experience as a clinician and researcher and extends it with five key training goals: 1) Measuring outcomes for OUD-related care across systems, 2) Advanced qualitative inquiry, 3) Implementing health systems change, and 5) Manuscript and grant writing. With successful completion of this project, training activities, and mentorship from a team of experienced investigators, Dr. Lowenstein will be well-prepared to lead an independent research agenda designed to improve care and outcomes for people with OUD.

NIH Research Projects · FY 2026 · 2022-06

Project Summary When sick with an infectious, autoimmune, or neoplastic disease, humans report sleepiness and excess or unrefreshing sleep. The features of human sleep during sickness are shared with those in Drosophila. Sickness behavior in fruit flies as in humans and other species is induced by stressors such as bacterial infection or aseptic injury, which reduce activity and increase sleep. While the causal mechanisms underlying altered sleep during sickness are unknown, immune system and lipid dysregulation are implicated. We will exploit the Drosophila genetic model to directly test specific interactions between the innate immune system and lipid homeostasis to determine how these interactions contribute to sleep need during sickness. We recently identified a novel brain-expressed antimicrobial peptide (AMP) nemuri (nur), which links the immune response with sleep. nur is strongly induced by infection and sleep deprivation and loss of it reduces sleep under these conditions. Our preliminary data indicate that induction of nur is mediated by Nuclear Factor κB transcription factors (NFκB). Overexpression of nur in the brain also promotes sleep and is associated with dysregulation of lipid metabolism in a manner that parallels that of patients with myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS). Preliminary data also indicate lipid depletion by sustained sleep deprivation. Based on these findings, we hypothesize that prolonged expression of nemuri, either genetically or as induced by chronic sleep deprivation or infection, causes lipid bilayer stress (LBS). Under normal circumstances, the unfolded protein response, mediated by IRE1/XBP1 signaling, resolves LBS (UPRLBS) and terminates the acute stress-induced sleep response. However, prolonged activation of nur disrupts NFκB activity, which is central to the innate immune response. The suppression of NFκB signaling subsequently prevents an appropriate response to LBS by reducing the adaptive lipid homeostatic UPRLBS, thereby increasing sleep need. We will use behavioral genetic, lipidomic, biochemical, and pharmacological approaches to test key aspects of this hypothesis. Given that immune, lipid metabolic, and UPR pathways are highly conserved between flies and mammals, results of our proposed studies will provide a framework for understanding a mechanism for sleep during sickness, with implications for unrefreshing sleep and fatigue associated with numerous illnesses in humans.

NIH Research Projects · FY 2025 · 2022-06

PROJECT SUMMARY Diseases of the biliary system comprise a significant percentage of the indications for liver transplantation in adult and pediatric populations. The pathogenesis of these disorders likely involves cellular responses to the harmful effects of bile, to which epithelial cells lining the bile ducts are continuously exposed. Bile toxicity arises largely from the oxidative damage caused by bile salts, whose detergent properties disrupt intracellular organelles, particularly mitochondria. Bile duct epithelial cells, also known as cholangiocytes, have evolved strategies for preventing bile toxicity in addition to having robust anti-oxidant defense systems, which are present in all cells to combat reactive oxygen species generated during normal metabolism. Work from my laboratory using the zebrafish system has identified regional susceptibilities of cholangiocytes to redox stress imparted by the plant toxin biliatresone, whose consumption is associated with epidemic biliary atresia (BA) in livestock. Our most recent studies indicate a comparable role for regional variantion in heat shock mediated proteostasis in cholangiocytes exposed to biliatresone, and that these responses can be modified by the activation of cGMP signaling. Importantly, there is compelling clinical data indicating the modulation of stress responses contributes to risk and outcomes in human BA, thus validating the biliatresone models we employ. The goal of this proposal is to study the mechanisms responsible for the variation in cholangiocyte stress responses using zebrafish, mouse and human cell culture models. In Aim 1, we will characterize the molecular determinants of the redox stress response in zebrafish intra-hepatic and extra-hepatic cholangiocytes, determine whether variation in these responses evolve from epigenetic factors that arise during biliary development, and correlate these findings with mammalian cholangiocyte biology using mouse models. In Aim 2, we will identify the cellular targets responsible for biliatresone-induced injury and stress in mammalian cholangiocytes using two approaches; first, via pull-down experiments using photo-activatable biliatresone analog; second, by conducting a genome-wide CRISPR positive selection screen for genes required for biliatresone toxicity. In Aim 3, we will explore links between cGMP signaling and proteostasis and how this affects susceptibility of cholangiocyte to injury using three approaches: first, by defining the downstream regulators of cGMP signaling that mediate its inhibition of biliatresone toxicity, second, by correlating these data with a comprehensive analysis of the cholangiocyte proteome, and how it is modified by biliatresone and cGMP signaling, third, defining the sub- cellular domains of cGMP signaling activity and its downstream mediators.

NIH Research Projects · FY 2026 · 2022-06

Although there is a rich history relating single-neuron spiking activity to brain function, our understanding of the neural bases of perception and behavior depends on identifying information processing within neuronal circuits that operate over different spatial scales. Despite their importance as a canonical model of information flow, our understanding of whether and how such bidirectional hierarchically organized circuits contribute to auditory perception remains elusive, especially in primate models of hearing. The overarching goal of this Roi proposal is to fill this knowledge gap by identifying bidirectional, hierarchically organized circuits that contribute to auditory perception. We will use predictive coding (PC) -a highly influential theoretical framework for auditory statistical learning and perceptual inference- as an experimental lever to test whether its predictions are instantiated in cortical circuits of the ventral auditory pathway. We focus on the ventral pathway because, in rhesus monkeys, this pathway contributes causally to auditory perception. To test the circuit predictions of PC, we insert multi-contact electrode(s) orthogonally to the cortical laminae and record single-unit and neuronalpopulation activity in the core auditory cortex and in the ventral auditory pathway While recording, rhesus monkeys participate in passive listening paradigms and an active listening task. During passive listening, monkeys listen to sequences of auditory stimuli containing both predictable and unpredictable events, which allow probes of prediction-error and prediction-related activity. During active listening, monkeys implicitly learn to use auditory cues to predict the onset of an auditory target; this task is a direct behavioral assessment of the strength and temporal dynamics of auditory statistical learning. In Specific Aim #1, we identify the interlaminar circuitry underlying PC and its hierarchical organization along the ventral auditory pathway. In Specific Aim #2, we identify the feedforward and feedback circuitry between the prefrontal and auditory cortices that underlies PC. The findings from this proposal will provide invaluable insights into the functional organization of cortical circuits underlying PC and statistical learning along the ventral auditory pathway. In turn, these insights may lead to the formulation of testable hypotheses regarding the neurophysiological underpinnings of deficits in statistical learning that have been identified in clinical conditions such as autism and schizophrenia.

NIH Research Projects · FY 2025 · 2022-06

Bicuspid aortic valve (BAV) repair is a promising surgical treatment for young adults with aortic regurgitation (AR). However, BAV repair surgery remains underutilized and variably applied across institutions, owing in part to the lack of a standardized approach to BAV repair planning. Currently, BAV repair planning relies primarily on intraoperative manual measurements of the valve made by direct observation while the heart is in an arrested state, making it difficult for the surgeon to identify defects in valve dynamics under physiological conditions. To address this challenge, the long-term goal is to develop a multimodal 4D image analytics and valve modeling platform that systematically characterizes pre-operative BAV morphology and dynamics and enables patient-specific surgical planning. The overall objectives of this proposal are to (i) fill a knowledge gap in the precise anatomical relationships between the aortic cusps, annulus, and root that make a BAV functionally competent, and (ii) develop computational image analytics to precisely identify the patient- specific, anatomical and dynamic distortions that cause AR so that these defects can be prioritized for risk stratification and planning of BAV repair surgery. This work will be carried out by pursuing three specific aims: (1) Design and assess an automated segmentation and modeling algorithm for 4D reconstruction of the BAV apparatus from multiple clinical imaging modalities; (2) characterize the morphological and dynamic features of BAV competence and create a machine learning method for comprehensive anomaly detection in regurgitant BAVs; (3) evaluate a BAV repair planning system using images acquired from valve repair procedures at three institutions. The proposed project leverages the complementary benefits of two modalities: real-time 3D transesophageal echocardiography and 4D computed tomography angiography, which capture both the morphological detail of the aortic cusps with high spatial resolution and the motion of the 3D BAV apparatus with high temporal resolution. The innovation of this project is that the proposed tools could change how BAV repair planning is carried out. Instead of relying on intraoperative inspection of the valve while it is unpressurized, the surgeon can interactively visualize image-derived BAV models and quantify dynamic mechanisms of AR when the valve is in a pre-operative 4D physiological state. The significance of this research is that it could promote consistency in valve repair planning across institutions, decrease surgeons’ reliance on intuition and trial-and-error, and thereby increase the utilization of BAV repair in young adults. This would have quality of life advantages relative to conventional valve replacement, which requires lifelong anticoagulation therapy (mechanical valves) or multiple re-replacements due to limited durability (bioprosthetic valves). Ultimately, the systematic analysis of multimodal image data for computer-aided valve defect detection will broadly benefit advancement of surgical treatments for acquired and congenital heart disease.

NIH Research Projects · FY 2026 · 2022-06

Abstract The current state of magnetic resonance imaging (MRI) methods in neurooncology offers great potential for providing rich characterizations of structural, physiological, and metabolic character- istics of brain tumors, especially gliomas, which are complex and highly heterogeneous cancers. Glioblastoma (GBM), in particular, has a grim prognosis, with median overall survival (OS) less than 15 months with relatively little improvement in the past 15 years since the Stupp protocol was introduced. Many experimental treatments are being pursued; however, OS has largely remained stagnant. Some of the obstacles in improving this outcome have been 1) disease heterogeneity, which both renders it difficult to detect treatment effects in Phase 1 or even Phase 2 trials, and calls for personalized, rather than one-size fits-all, treatment strategies; 2) methods used for tumor characterization based on size, enhancement, perfusion and diffusion properties are relatively crude and don't fully leverage the richness of imaging data or their spatial heterogeneity. Quanti- tative imaging and machine learning (QIML) methods developed in the past decade have shown great potential for dissecting the spatial, temporal and inter-patient heterogeneity of GBM; for discoveringrelationships between imaging and molecular characteristics ; foroffering personalized predictions of clinical outcome; and for leveraging subtle multi-parametric relationships in the data to detect peri-tumoral infiltration or distinguish treatment related changes, i.e., pseudo-progression (PsP), from true tumor recurrence. Our group has been at the forefront of QIML, with emphasis on a) obtaining rich imaging phenotypes relying on multi-parametric signals, texture parameters, shape properties, spatial patterns derived from atlas registration, and biophysical models of tumor growth, and b) integrating such imaging signatures using machine learning into predictors of clinical outcome, early recurrence from peri-tumoral infiltration, PsP, and radiologic subtypes of GBM. Despite their promise, QIML methods have a notorious limitation: they might overfit specific datasets from which they are derived, and might display poor reproducibility under real-life conditions of variable scanner types and imaging protocols. In this proposal we aim to leverage the recently formed ReSPOND (Radiomics Signatures for PrecisiON Diagnostics) consortium, to integrate, harmonize, and analyze 4,578 datasets from 14 centers around the world, and hence more appropriately train and cross-validate QIML tools for a wider generalizability. This consortium will generate an unprecedented database of diverse and carefully harmonized sets of MRI and clinical measures, and aims to provide the community with robust and reproducible QIML models contributing to precision diagnostics and personalize treatment for this dreaded brain cancer.

NIH Research Projects · FY 2024 · 2022-06

PROJECT SUMMARY: Each year 13- to 24-year-olds disproportionately compose the number individuals diagnosed with human immunodeficiency virus (HIV) in the United States. Preexposure prophylaxis (PrEP), a once daily antiretroviral regime, is an effective method to prevent the transmission of HIV in adolescents at substantial risk for acquiring HIV, however, the effect of this regimen on the development of critical brain structures during adolescence is unknown. Adolescents taking PrEP are uniquely vulnerable to myelin impairments as the adolescent brain is undergoing high rates of myelination. Our lab has shown that primary oligodendrocyte precursor cell cultures treated with therapeutic concentrations of select antiretroviral drugs displayed dose-dependent decreases in oligodendrocyte maturation. A gap in our knowledge is the mechanistic basis of the inhibition of oligodendrocyte maturation by antiretrovirals in an HIV-negative, adolescent population. My preliminary data suggests that emtricitabine (FTC) and tenofovir disoproxil fumarate (TDF), the drugs composing PrEP, de-acidify oligodendrocyte lysosomes. Furthermore, my preliminary data also suggests that PrEP decreases SREBP2 expression in oligodendrocytes in vitro. mTORC1 regulates lipogenesis through the transcription factors SREBP 1 and 2, with the latter regulating the expression of genes involved in cholesterol synthesis, the rate-limiting step of myelination. Lysosome de-acidification has been shown to result in increased mTORC1 signaling in osteoclasts but remains to be investigated in oligodendrocytes. Additionally, overactivation of mTORC1 in oligodendrocytes is known to result in hypomyelination and decreased SREBP2 expression. Taken together, I hypothesize that oligodendrocyte maturation is inhibited by PrEP through lysosome deacidification resulting in increased mTORC1 activation and decreased lipogenesis. I will address this hypothesis in the following specific aims. In Aim 1 I will demonstrate oligodendrocyte maturation is impaired by PrEP through lysosome de- acidification in vitro. In Aim 2, I will demonstrate that PrEP inhibits oligodendrocyte maturation through increased mTORC1 signaling and subsequent decreased lipogenesis in vitro. In Aim 3, I will demonstrate that oligodendrocyte maturation and myelination are impaired by PrEP and lysosome acidification rescues myelination in vivo. Overall, these experiments will investigate the previously unanswered question of whether PrEP affects oligodendrocyte maturation in HIV- negative individuals.

NIH Research Projects · FY 2025 · 2022-06

PROJECT SUMMARY Alzheimer’s disease and related dementias (ADRD) are a leading cause of neurological disability, causing devastating impairments in cognition and functional capacity . Although there has been considerable progress in identifying mechanisms underlying ADRD, and symptomatic treatment is available for some ADRD, there is urgent need for greater understanding of the mechanisms underlying these dementias to identify potential treatment targets and to contribute to successful treatment trials. Because of its ability to resolve brain structure and function noninvasively, neuroimaging has become a leading strategy used for translational research in ADRD and a prominent biomarker strategy for clinical differential diagnosis of ADRD, for defining preclinical stages of ADRD, and for evaluating for target engagement and therapeutic responses in trials. Modern neuroimaging is an inherently interdisciplinary field combining expertise in physics, chemistry, engineering, computer science, neuroscience, and statistics to localize and quantify a large array of metrics reflecting the molecular, cellular, structural, functional, and physiological properties of the central nervous system. Additional expertise in topics such as genomics, proteomics, metabolomics, neuropharmacology, psychology, and neuromodulation are also required to optimally combine or apply neuroimaging with other key areas in ADRD research. Finally, because of its multidimensional nature, neuroimaging is a “big data” field requiring expertise in informatics and data management. The University of Pennsylvania has a longstanding track record of innovation and translation in biomedical neuroimaging in ADRD, and supports state-of-the-art instrumentation for in vivo MRI, PET, CT, and optical imaging in both humans and preclinical models as well as extensive expertise and resources in image processing and spatiotemporal statistics. In response to PAR-21-112 the proposed “Cross-disciplinary training program in translational neuroimaging of ADRD” will draw predoctoral and postdoctoral trainees from both biological and quantitative science backgrounds who are seeking a research career focusing on neuroimaging in ADRD. A core curriculum covering neuroimaging methods will be combined with specializations in 1) imaging studies of disease mechanism, 2) imaging biomarkers and neuroimaging trials, or 3) statistical and computational approaches to neuroimaging data. A diverse faculty with outstanding track records of research and mentoring in neuroimaging and related fields will participate, with each trainee receiving dual mentorship from faculty with complementary expertise. The aims of the proposed training program are 1) to train the next generation of ADRD neuroimaging researchers, 2) to increase diversity in neuroimaging research through efforts to increase diversity in both program trainees and program faculty, 3) to expand local ADRD neuroimaging research through participation of training faculty with related domain expertise, and 4) to provide training in the Responsible Conduct of Research and Experimental Rigor, particularly as it applies to neuroimaging.

NIH Research Projects · FY 2026 · 2022-06

Project Summary Chemotherapy resistance remains a major barrier to successful treatment of patients with acute myeloid leukemia (AML), contributing to high rates of relapse and mortality. Development of more effective treatments for AML is imperative, particularly therapies with alternative mechanisms of action to circumvent chemoresistance. Adoptive cellular immunotherapy using CD19-targeting chimeric antigen receptor (CAR)- expressing T cells has drastically improved the treatment of patients with multiply relapsed/refractory B-cell lymphoblastic leukemia (B-ALL) and lymphoma and was approved by the FDA. However, successful translation of AML immunotherapies has lagged behind and remains a significant unmet medical need. To date, CAR T cells targeting CD33 or CD123 for AML have shown potent preclinical anti-AML activity, but also induced severe myelotoxicity via on target/off tumor damage to hematopoietic stem cells (HSCs). We have developed an innovative system to isolate single-domain nanobodies (Nb) that preferentially bind AML cells and enable cognate CAR T cells to kill the cancer cells. One of these nanobodies, Nb157, specifically binds to the cell surface protein CD13 (aminopeptidase N), which is often upregulated in adult AML specimens and leukemia stem cells (LSCs). In preliminary studies, we demonstrated that Nb157/CD13 CAR T cells (CD13CARTs) potently eradicated AML cells in preclinical animal models. TIM-3, an inhibitory receptor of certain immune cells, is upregulated in AML blast cells and LSCs, but not expressed in human HSCs. Thus, we generated the 1st generation bispecific and split CD13/TIM-3 CARTs (1st G bCARTs) and demonstrated that the bCARTs potently eradicated AML cells in preclinical animal models, with significantly reduced toxicity to HSCs. To further improve the safety profile of the bCARTs, the 2nd generation bCARTs were generated and they did not induce obvious toxicity to HSCs in our ex vivo analysis. We hypothesize that further development of the bispecific or inducible bispecific CARTs can eradicate AML in patient-derived xenograft (PDX) models with little or tolerable off-tumor toxicity. Three specific aims are proposed to test this hypothesis. Aim 1 will evaluate the 2nd generation bispecific CD13/TIM-3CARTs (bCARTs) in maximizing selective AML killing. Aim 2 will investigate efficacy and specificity of inducible CD13/TIM-3CARTs (ibCARTs) in killing AML cells. Aim 3 will develop bispecific CLL-1/TIM-3 bCARTs to selectively killing AML cells. Results obtained from these studies are imminently translatable to the clinic in the near future for patients with relapsed/refractory AML.

NIH Research Projects · FY 2024 · 2022-06

PROJECT SUMMARY Sleep is an evolutionarily conserved behavior that is widely observed across the animal kingdom. It is characterized by transitions between different vigilance states: wake, rapid eye movement (REM) sleep, and non-REM (NREM) sleep. These transitions are controlled by interactions between different neuronal populations and are under the influence of homeostatic and circadian mechanisms. As sleep has many beneficial and restorative effects, good quality sleep is important for mental and physical health. It has been well-established that stress and sleep have a bidirectional relationship. Stress is known to be a major cause of disrupted sleep. Chronic sleep disruption can lead to an increased risk of developing psychiatric disorders. The paraventricular nucleus of the hypothalamus (PVN) contains corticotropin-releasing hormone (CRHPVN) neurons that have been shown to be activated by stress. Central and systemic administration of CRH has been found to induce wakefulness. Additional studies have found that central blockade of the CRH receptor 1 (CRHR1) reduces the wake-promoting effects of CRH injection, suggesting this receptor plays a key role in CRH-mediated arousal. However, the neural mechanisms by which CRH neurons regulate wakefulness are not very well understood. Tracing studies have revealed that CRHPVN neurons project to the preoptic area of the hypothalamus (POA), a well-known sleep center containing neurons that are crucial for sleep regulation. In situ hybridization studies have shown that CRHR1 is expressed in the POA. While CRH has been implicated as a regulator of wakefulness, the role that CRHPVN neurons and their projections to the POA (CRHPVN→POA) play in the sleep-wake cycle and sleep homeostasis has not been fully investigated. The central hypothesis of this proposal is that CRHPVN neurons control wakefulness and impair the homeostatic response to sleep pressure following chronic stress. To address this hypothesis, this proposal will use a genetic mouse model, CRH-Cre, to specifically label CRH neurons, in vivo calcium imaging, optogenetic manipulations, CRISPR-Cas9 gene editing techniques, and a chronic social defeat stress paradigm to manipulate these neurons and their projections. Aim 1 will determine the role of CRHPVN→POA projections in regulating sleep and wakefulness. Aim 2 will investigate the role of CRHPVN neurons and CRHPVN→POA projections in sleep homeostasis following chronic stress. Understanding how CRHPVN neurons promote wakefulness and regulate sleep homeostasis in response to chronic stress will further elucidate how the circuits controlling stress are interconnected with those regulating sleep and wakefulness.

NIH Research Projects · FY 2025 · 2022-06

PROJECT SUMMARY Millions of Americans living with serious illness experience burdensome symptoms and receive aggressive care that is not aligned with their goals and preferences. A growing body of evidence suggests that palliative care, which entails a supportive approach to care focused on maximizing quality of life, improves patient- centered, clinical, and economic outcomes. For this reason, national guidelines recommend that clinicians either provide palliative care themselves (primary) or consult experts (specialist) as part of standard serious illness care. For these reasons, most hospitals in the U.S. have invested in specialist palliative care programs. Yet, palliative care delivery remains insufficient among patients with serious illness, particularly those with advanced Alzheimer's Disease and Related Dementias (ADRD), and use of specialist palliative care services is often inefficient and inequitable, largely due to clinicians' difficulty identifying which patients are most likely to benefit from them. Many hospitals have begun to implement prognostic triggers in the electronic health record (EHR) to facilitate more reliable and equitable patient identification, however, none have been rigorously tested for their effects on patient-centered outcomes. Furthermore, palliative care triggers cannot solely rely on the limited workforce of palliative care specialists, but rather approaches that promote primary and specialist palliative care are needed, yet evidence is lacking for how to optimally do so. The main objective of this study is to evaluate a strategy that combines an EHR-based prognostic-trigger with two effective clinician-directed nudges to provide either primary or specialist palliative care for seriously ill hospitalized patients. Specifically, the behavioral intervention involves a simple EHR alert to the primary clinicians caring for identified patients that requires them to actively choose whether or not to provide primary palliative care, and only if they decline, a default order for specialist palliative care is entered from which they can opt-out. We will conduct a hybrid type 1 pragmatic, cluster randomized trial among nearly 7,000 patients across 6 diverse hospitals to study the intervention's effectiveness on hospital-free days and numerous other patient-centered, clinical, and economic outcomes. We will also conduct an embedded mixed methods study to understand clinician and hospital contextual factors that influence the intervention's uptake. Finally, we will evaluate for treatment effect heterogeneity among patients with ADRD and other pre-specified subgroups to determine which types of patients derive the greatest benefit from a systematic approach to nudge palliative care. This study will provide high-quality evidence regarding the effectiveness of a scalable and sustainable approach to promote collaborative primary and specialist palliative care among a large and diverse patient cohort, will advance the science of triggers for palliative care, will provide new insights into the types of patients most likely to benefit from systematic identification for palliative care, and will create new knowledge about how to establish hospital environments conducive to desired clinician behavior change to improve serious illness care.

NIH Research Projects · FY 2025 · 2022-06

Abstract: MPI R01- Hambardzumyan and Becher Pediatric high-grade gliomas (pHGG) account for the most cancer-related deaths in children and have a median survival of 12-15 months. One promising avenue of research is developing novel therapies targeting the properties of non-neoplastic cell types within the tumor, such as myeloid cells, including tumor-associated macrophages (TAMs) and neutrophils. Much is known about TAMs in adult high-grade gliomas. However, very little is known about TAMs and neutrophils in pHGGs. pHGGs more commonly arise in infratentorial locations like the brainstem, beyond the cerebral hemispheres as seen in adults. pHGGs also harbor distinct histone mutations not found in adults. This raises the question of whether pHGGs possess a distinct constituency of TAMs due to their unique genetic landscapes and locations. Using human pHGG tissue samples and NanoString RNA sequencing, we demonstrate brainstem/midline pHGGs (DMG) and murine diffuse intrinsic pontine glioma (DIPG) possess higher inflammatory scores and increased neutrophil scores compared to hemispheric pHGGs, which are associated with shorter patient survival. When examining only human hemispheric pHGGs, our results revealed two patient subsets with high and low inflammatory scores. Patients with a high inflammatory score had significantly shorter survival times compared to those with low inflammatory scores. We also show that human pHGGs possess high infiltration of IBA1+ TAMs, which are the most abundant non-neoplastic component of the pHGG tumor microenvironment (TME). Our preliminary data utilizing mouse models to recapitulate pHGG in newborn immunocompetent mice combined with various histone mutations in biologically relevant locations demonstrate murine tumors are strikingly similar to their human counterparts with regard to their inflammatory immune profile and myeloid cell infiltration. Using PDGFB and PDGFA as driver mutations to generate hemispheric pHGG in mice, we were able to recapitulate human pHGGs with high and low inflammatory scores. We showed that in comparison to PDGFA-driven tumors, PDGFB tumors have a higher inflammatory score, increased infiltration of monocytes from the blood, and shorter survival time of tumor-bearing mice. We identify CCL3 as a potential key chemokine for CCR1/CCR5-positive monocytes and CXCL1 for CXCR2-positive neutrophil recruitment in pHGG. Together, these results provide strong rationale to extend our studies to understand how specific histone mutations and tumor locations influence myeloid cell infiltration and how these cells promote pHGG growth. The outcome of these studies will: (i) reveal the molecular and functional diversity of the myeloid compartment of pHGG; (ii) determine how distinct myeloid subsets and myeloid-specific genes influence tumor growth and the TME; (iii) provide insight into how myeloid subsets influence, and are affected by driver mutations, pediatric-specific histone mutations, and tumor location (hemisphere versus brainstem). This study will also determine whether targeting monocyte and neutrophil infiltration will be a viable therapeutic avenue for exploitation in pHGG.

NIH Research Projects · FY 2025 · 2022-06

More than 1/3 of the world's 65 million people with epilepsy (~3.3 million in the U.S.) have seizures that cannot be controlled by medications. Surgery and implanted devices are options for many, but their success depends upon manually mapping epileptic networks, which is only possible for some patients, and poorly standardized. When surgical targets are identified, there is currently no rigorous way to select the best surgical approach. The overall aim of this proposal is to develop rigorous, standardized, quantitative methods to: (1) map epileptic networks from imaging and Stereo EEG (SEEG), (2) pick the best region for resection, ablation or neuromodulation for individual patients from their data and clinical hypotheses, and (3) to determine when focal intervention is unlikely to succeed. These methods would have tremendous positive impact on clinical care. Over the past four years we have made substantial progress towards these goals. We have developed: (1) robust measures derived from subdural intracranial EEG (ECOG) that predict outcome from epilepsy surgery; (2) personalized methods that localize epileptic networks and predict the impact of different interventions on seizure control; (3) tools that predict the path of seizure spread from combined MRI and IEEG. We also have a track record of openly sharing our methods, data, results and code on http: //ieeg.org, to accelerate research. Based upon this work, we now innovate to solve 3 fundamental challenges to translating our work into practice: (1) Guiding SEEG: We must develop new methods that account for the sparser sampling and different philosophy of stereo EEG, which maps a network of connected brain regions and tests clinical hypotheses about where seizures initiate and propagate; (2) Assessing sampling bias and missing information: We will develop methods to determine if electrodes sample all key regions of the epileptic network, to ensure we do not falsely localize due to missing information; (3) Validating in a larger population across centers: In parallel to refining the above methods, we will validate and harmonize our analyses across centers in a large number of patients to harden it for clinical use. In a novel model, we have engaged a group of major surgical epilepsy centers to openly collaborate, standardize methods, aggregate data, and share all algorithms, computer code, data and results on http: //ieeg.org. Our central hypothesis is that our quantitative methods can be standardized across centers, predict outcome from personalized epilepsy surgery, and ultimately be translated to improve clinical care. This work is significant because it merges state of the art network neuroscience, engineering, neurology and neurosurgery to make practical tools to improve and standardize patient care. It also establishes a collaboration between 15 major epilepsy centers to standardize and share data. Finally, this project leverages a thriving collaboration between experts in neurology, computational neuroscience, neurosurgery, neuroimaging and bioengineering at Penn, with a strong track record of clinical translation.

NIH Research Projects · FY 2026 · 2022-06

Rare genetic causes of human disease have the potential to reveal mechanistic insights into more common sporadic disease. Tauopathies are a group of fatal neurologic diseases, including Alzheimer's disease, where dementia and neurodegeneration are the result of accumulation of pathologic tau protein aggregates in the form of neurofibrillary tangles, Pick bodies, or glial inclusions. We have identified a novel autosomal dominant form of frontotemporal dementia with tau inclusions associated with a novel hypomorphic genetic mutation in VCP which may be linked to a loss of anti-tau disaggregase activity. However, hypermorphic VCP mutations cause a distinct disease called multisystem proteinopathy which can manifest as frontotemporal lobar degeneration with TDP-43 inclusions. We propose three specific aims to understand the basic molecular mechanisms by which this gene mutation leads to diverse pathologies. We will perform structural biology studies to better understand how VCP protein interacts with pathologic protein aggregates, and how VCP mutations affect disaggregase activity. We will extend these studies into cellular/neuronal VCP knock-in models to determine how VCP mutations affect cellular VCP activity within a cellular context. Finally, we will determine whether gains or losses of VCP activity can modify tau toxicity in vivo. Together, these mechanistic studies will elucidate basic mechanisms by which VCP dysfunction leads to different types of proteinopathy, providing the basis for future novel anti-tau therapies based on modulating VCP activity.

NIH Research Projects · FY 2025 · 2022-06

PROJECT SUMMARY Congenital lung diseases, such as inherited surfactant protein syndromes, cystic fibrosis, and alpha-1 antitrypsin deficiency, are a significant source of pediatric morbidity and mortality. Treatment options for neonatal patients with lung disorders that present with respiratory failure are limited to palliative care or pediatric lung transplant. As such, there is a clear clinical demand for new therapies that allow for early correction of congenital lung diseases to reduce pediatric morbidity and mortality. Recent advances in gene editing technologies, such as CRISPR-Cas9 systems, have unlocked the potential to correct pathogenic mutations and thereby treat congenital disorders at their source. Performing gene editing in utero offers the added benefits of reversing genetic abnormalities prior to the transition to postnatal life, when pulmonary function becomes essential, and harnessing normal developmental properties of the fetus for more efficient correction. Traditionally, viral vectors have been used to study in utero gene therapy in animal models. Although these studies are encouraging, discovery of alternative, potentially safer, delivery vehicles will advance the field toward clinical translation. Thus, this proposal aims to investigate the potential of ionizable lipid nanoparticles (LNPs), a promising non-viral delivery platform, for nucleic acid delivery to the mouse fetal lung. Fetal lung optimized lipid nanoparticles (FLO-LNPs) will be generated through a multi-stage optimization scheme. In Aim 1, a diverse library of 24 ionizable lipid structures will be screened to identify the ionizable lipid that best delivers mRNA to the fetal lung. In Aim 2, LNP formulations will be optimized using a Design of Experiments scheme for minimal toxicity and maximal delivery of a CRISPR-Cas9 systems in mouse precision cut lung slices. In Aim 3, the optimized LNP formulation will be modified via antibody conjugation and tested for cell-specific targeting in the fetal mouse lung. Ultimately, this proposal – conducted as an interdisciplinary project between sponsors in the Department of Bioengineering, Perelman School of Medicine, and Children’s Hospital of Philadelphia at the University of Pennsylvania – will allow for the development of a novel LNP delivery platform that can be applied in subsequent work to deliver in utero gene therapies for congenital lung disease.

NIH Research Projects · FY 2025 · 2022-06

Project Summary/Abstract: Alzheimer's disease and Alzheimer's disease-related dementias (AD/ADRD) relentlessly erode affected individuals' decision-making abilities. As these abilities decline, persons with AD/ADRD often engage another person, typically their care partner, in decision making. In clinical encounters, the addition of a third party, the clinician, further complicates communication and decision making. There is a critical need to identify interventions with the potential to improve dyadic and triadic communication. Supported decision making is such an intervention. In supported decision making, an adult with impaired decisional abilities enters into a structured agreement with another person; this agreement identifies domains in which the adult with impaired decisional abilities needs and wants decision-making help, and it specifies the kinds of help sought. This agreement then guides the supported decision-making process. For example, based on the agreement, this other person may talk through the pros and cons of various treatment options to aid the adult with impaired decisional abilities in reaching a decision. Supported decision making acknowledges the potential vulnerabilities of persons with impaired decisional abilities while also promoting their ability to engage in self- determination and, in turn, their wellbeing. Supported decision making is increasingly being used with young adults with intellectual and developmental disabilities (I/DD) who reach the age of majority (i.e., are no longer minors under parental care), and evidence suggests that supported decision making promotes their wellbeing. Moreover, many states are beginning to recognize supported decision making as an option for persons with impaired decisional abilities. Yet, no extant supported decision-making resources are AD/ADRD-specific, and there is a lack of evidence on the outcomes of supported decision making for persons with AD/ADRD, their care partners, or clinicians. In Aim 1, we will modify existing supported decision-making tools for use in AD/ADRD with input from Delphi panelists including patients, care partners, clinicians, and experts in consent, capacity, and supported decision making. In Aim 2, we will interview patients, care partners, and clinicians to understand the attitudinal, normative, and self-efficacy beliefs associated with greater intention to use the AD/ADRD-specific supported decision-making toolkit, as well as perceived barriers to its use. In Aim 3, we will pilot test the AD/ADRD-specific supported decision-making toolkit with patients, care partners, and clinicians at the Penn Memory Center and assess decision making following delivery of the intervention relative to usual care. In order to develop potent, scalable behavioral interventions to meet the medical decision-making needs of the millions of Americans with AD/ADRD, we must increase our understanding of the mechanisms underlying supported decision making and assess its ability to improve their wellbeing.