University Of Pennsylvania

NIH Research Projects · FY 2024 · 2023-09

Project Summary Acute myeloid leukemia (AML) is a lethal blood cancer characterized by a differentiation “block” that prevents myeloid progenitor maturation resulting in uncontrolled proliferation. Chemotherapy, the current standard of care, is often ineffective and can result in cytotoxicity and relapse. New treatment options are desperately needed to treat AML’s poor prognosis. Differentiation therapy is a novel method that aims to reactivate latent maturation programs and induce cell cycle exit. This therapeutic strategy is curative in the promyelocytic (APL) AML subtype but underexplored in other AMLs. Epigenetic factors help sustain the characteristic AML differentiation block. The demethylase LSD1 has emerged as a promising target for differentiation therapy. Pharmacologic inhibition of LSD1 (LSD1i) induces cellular differentiation in many AML subtypes. However, the extent of differentiation varies between AML models, with a modest effect in aggressive AML models. Therefore, LSD1i will not induce terminal differentiation as a monoagent treatment. We hypothesized that targeting additional epigenetic regulators simultaneously with LSD1i may induce complete, terminal differentiation and lead to disease remission. To identify potentiators of LSD1i, we conducted CRISPR gain-of-differentiation screens with a chromatin-focused sgRNA library in multiple AML cell models with or without LSD1i co-treatment. These screens unveiled a synergistic induction of differentiation when KO of MEN1 is combined with LSD1i. I confirmed that combinatorial small molecule inhibition of LSD1 and MEN1 induces differentiation and reduces proliferation most commonly in MLL-rearranged AMLs, and to a lesser extent in selected MLL-wild type AMLs. This proposal has two aims: First, I will test the therapeutic potential of targeting MEN1 in combination with LSD1 inhibitors in primary patient samples ex vivo and in patient-derived AML transplant models. Patient samples will be treated with a MEN1 inhibitor in combination with LSD1 inhibitors, and effects on differentiation and proliferation will be quantified. Secondly, I will determine the epigenomic mechanisms by which MEN1 and LSD1 inhibition synergize to induce terminal differentiation, and then test whether these mechanisms are conserved in primary patient samples.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY/ABSTRACT Despite its critical role in meal cessation, the neuroendocrine mechanisms underlying sensory-specific satiety (SSS) are largely unknown. SSS is the transient reduction in motivation and pleasantness for a recently consumed food. While this selective decrease in hedonic value suppresses consumption of the same food, in leads to a comparative increase in the reward value of uneaten palatable foods. As a result, a variety of readily available alternative foods can drive overconsumption. Given the variety of highly caloric foods pervasive in the modern environment, it is important to understand the mechanisms by which SSS regulates food intake. To this end, we propose to use our novel rat model of SSS to investigate the neural action of one satiation signal, glucagon-like peptide-1 (GLP-1). Indeed, our preliminary data indicate GLP-1 receptors (GLP-1Rs) in the hindbrain are critical for the SSS-induced decline in consumption of the same food. Moreover, hindbrain GLP- 1Rs are sufficient to prevent the overconsumption of an available alternative food. These data show hindbrain GLP-1 signaling is important for decreased consumption of the same food. In contrast, the effects of GLP-1 signaling is muted during the comparative increased intake of an alternative available food. However, these data unearth additional questions regarding the neural underpinnings of SSS. Notably, the neural population(s) mediating this GLP-1 signaling are unknown. An integrator of peripheral signals, the nucleus tractus solitarius (NTS) of the caudal brainstem is well-positioned to host the GLP-1 signaling involved in SSS. Not only does the NTS contain GLP-1R-expressing cells, but it also holds preproglucagon (PPG) neurons that synthesize GLP-1. Therefore, in Aim I, we propose to investigate the role of these two distinct NTS neural populations in controlling SSS. Furthermore, it is unclear how the effects of GLP-1 signaling are muted to enable the increased intake of an alternative food. Two subcortical structures, the bed nucleus of the stria terminalis (BNST) and the central amygdala (CeA) are functionally and anatomically suited to modulate GLP-1 signaling. Both regions mediate aspects of ingestive behavior and reward, and each region projects directly to PPG NTS neurons. Thus, in Aim II we will characterize the role of BNST/CeA → PPG projections during increased intake of an alternative food. Considering the ever-growing prevalence of obesity in the United States, understanding how neuroendocrine signals mediate the impact of recent food experiences on consumption is an important step in the development of approaches for treating diseases of maladaptive ingestive behavior. To this end, our experiments will investigate central GLP-1-mediated mechanisms by which SSS modulates food intake.

NIH Research Projects · FY 2025 · 2023-08

PROJECT ABSTRACT Transesophageal echocardiography (TEE) is an ultrasound-based cardiac imaging procedure frequently used to facilitate informed surgical decision making and manage intraoperative complications during the quarter million cardiac surgeries performed in the US each year. And yet, there is no randomized evidence to support the use of intraoperative TEE in any type of cardiac surgery. This lack of randomized evidence is particularly problematic in isolated coronary artery bypass graft (CABG) surgery because observational comparative effectiveness studies are conflicted regarding the association between improved postoperative outcomes with (vs without) intraoperative TEE. Complicating matters, TEE is an invasive procedure where its unconfirmed clinical benefits may not outweigh its confirmed risks of gastroesophageal injury. The questions surrounding the unclear clinical efficacy, unknown real-world effectiveness, and undefined risk-benefit ratio of intraoperative TEE during isolated CABG surgery is a critical evidence gap with clinical equipoise that has resulted in widespread practice pattern variability and a near 50/50 split in TEE (vs no TEE) intraoperatively. Thus, the overall objectives of this K23 research will leverage this existing clinical equipoise and practice pattern variability to generate the required foundational data necessary to inform a future, multicenter, randomized controlled trial (RCT) that could resolve the questions surrounding the clinical efficacy, effectiveness, and risk- benefit ratio of intraoperative TEE use (vs lack of use) during isolated CABG surgery. To that end, three aims will test the central hypothesis that among a routine isolated CABG surgery population (i.e. a population suitable for randomization given the absence of valve disease), the treatment effects of intraoperative TEE on postoperative clinical outcomes after isolated CABG surgery will vary among subgroups (aim 1) and that randomizing routine isolated CABG surgery patients to either default (i.e. obligatory) or as-needed (i.e. backup) intraoperative TEE will garner adequate participation by stakeholders (aim 2), and demonstrate enough practical feasibility in a single-center, pilot RCT conducted at the Candidate’s institution (aim 3), to accomplish on a larger scale in a future, multicenter RCT. The information gained from completion of these three aims will provide the preliminary data required to inform subsequent, multicenter clinical trial(s) comparing perioperative clinical outcomes among routine isolated CABG surgery patients randomized to default vs as-needed intraoperative TEE. In combination with a structured course of mentoring, and a research training plan heavily focused on the how to plan, design, and conduct randomized clinical trials, this K23 will be the foundation for the Candidate’s progression toward becoming a leading clinical trialist and comparative effectiveness researcher dedicated to improving clinical outcomes in the cardiac surgical patient population.

NIH Research Projects · FY 2024 · 2023-08

Project Summary/Abstract The long-term goal of this project is to develop of a rapid, inexpensive, and sensitive screening diagnostic based on an innovative broad proteomic approach to improve our ability to diagnose infectious disease, monitor changes in the microbiome, measure the state of the host immune system, and identify disease specific biomarkers. Specifically, the initiative will provide proof of principle for a method to achieve simultaneous multi-pathogen detection and deep proteomic microbiome characterization. We will work to detect the presence of several hundred pathogens while also identifying thousands of microbes at clinically relevant levels. The short amino acid sequences or peptides from the proteins will provide markers that range from the virus variant level, such as SARS- CoV-2 variants of concern, to the family level, such as coronaviridae. In developing this diagnostic, a key challenge is to maximize the sensitivity of peptide marker detection while analyzing for many clinically relevant viruses and bacteria. We will therefore compare the performance of (a) a data-dependent acquisition method that searches a custom database and (b) a data-independent acquisition method that can identify thousands of proteins. The proposed study will begin with the analysis of approximately twenty model pathogens before proceeding to the more ambitious analysis of more than 230 oro-respiratory samples. We will assess the primary factors that determine the performance of each method and use our milestones to select one approach for further development. In the end, this research will demonstrate the feasibility of a broad protein-based screening diagnostic that can detect numerous pathogens and provide microbiome profiling to ultimately improve the diagnosis of disease and assessment of human health. The project team includes researchers with diverse expertise from Penn Engineering, Microbiology, and Penn Medicine.

NIH Research Projects · FY 2025 · 2023-08

Project Summary Functional status is defined by the ability to perform daily physical tasks necessary for independent living and is essential among persons with chronic kidney disease (CKD) for disease self-management, eligibility for kid-ney transplantation, and for ensuring good quality of life. People with CKD at all stages are at increased risk of poor physical functioning that occurs earlier than in the general population. Due to the high prevalence of phys-ical impairments among persons with CKD, the National Kidney Foundation clinical practice guidelines recom-mend regular assessments to identify those with physical limitations with the goal of improving outcomes. De-spite this acknowledgement of the importance of physical function to the health of persons with CKD, there has been limited research in the setting of CKD that would allow evidence-based clinical monitoring. To date, stud-ies in kidney disease populations have been limited by examining physical function at one time point, by evalu-ating relationships only with mortality, and by focusing largely on persons with kidney failure with replacement therapy (KFRT) and older adults. Early detection and intervention to prevent physical limitations and disability may be most effective if implemented during earlier stages of CKD and among younger adults. Risk factors of physical function decline have also been inadequately evaluated in persons with CKD and recommended physical function assessments are not routinely performed in CKD care. We propose to use nearly 20 years of follow up data from the NIDDK-sponsored Chronic Renal Insufficiency Cohort (CRIC) Study that represents diverse populations across the spectrum of CKD stages to enhance our understanding of the prognostic im-portance of physical function over time among persons with CKD, including insights to risk factors of physical function decline, and to build a clinical monitoring tool of physical function. We will characterize the trajectories of physical function along with their associated factors among people with CKD to increase the understanding of the pathways that lead to physical function decline and to identify modifiable factors for intervention develop-ment (Aim 1). We will determine the relationship of longitudinal physical function with onset of KFRT, hospitali-zations, and death (Aim 2). Lastly, we develop a monitoring tool for physical function that could guide treatment and prognostic decisions in CKD clinical practice. We will also test this tool in an external cohort with advanced CKD (Aim 3). Ultimately, the proposed study will increase our understanding of physical function in CKD, in-cluding insights into risk factors for decline, and its relationship to important outcomes; and the work will culmi-nate in a parsimonious set of physical function measures that can be used for clinical monitoring of physical health to inform treatment and prognostic decisions. This information could change how CKD patients are monitored in clinical care, guide patient-centered treatments, and promote broad improvement in physical func-tion for this high-risk population by catalyzing greater awareness and targeted interventions.

NIH Research Projects · FY 2025 · 2023-08

ABSTRACT High-grade gliomas (HGGs) are the most common primary brain malignancy in adults associated with very poor survival rates despite various treatments. Surgery is the current mainstay treatment for HGGs, and the main factor affecting survival rates (in over two decades) has been the increased extent of resection targeting the “visible” contrast-enhancing tumor (CET) seen on conventional contrast-enhanced MR imaging. However, since then research has shown that it is the “invisible” non-enhancing tumor (NET) which leads to progression or recurrence in HGGs by infiltrating the surrounding white matter (WM) tracts. This has led to the adoption of a supratotal resection (SpTR) approach, which includes resection of the `invisible' (microscopic) cancer beyond the visible contrast enhanced margins. SpTR has been shown to result in better patient outcomes with progression-free and overall survival. SpTR is undertaken using a combination of intra-operative techniques but having a pre-op assessment of the functional anatomy will enhance the chances of preserving function and maximizing tumor resection. Thus, the overarching goal of this Academia-Industry partnership (AIP) is to provide a treatment planning tool that will facilitate safe SpTR maximizing the benefit of surgical therapy while preserving neurologic function. The partnership builds on the technical expertise of UPenn for method development, the translational expertise of Synaptive to integrate into a clinically deployable product, and Mount Sinai's clinical expertise in evaluating it on patients. In Aim 1, UPenn will optimize and evaluate a tracking paradigm that provides enhanced visualization of WM fibers in NET. This will entail by combining tissue modeling, fiber tracking and tract delineation in clinically feasible multishell dMRI and optimize the paradigm for reproducibility and generalizability across patients and acquisitions. A comprehensive comparison of the approach to research and clinical paradigm will also be undertaken using retrospective data. The prototype for this tractography paradigm will be integrated into the Synaptive neuro-navigation product incorporating clinical and regulatory needs, with rigorous testing. The design will be optimized to maximize clinical utility assessed through a multi-surgeon evaluation across different Synaptive sites. This will culminate in the creation of an enhanced planning tool. Finally, in Aim 3, a prospective pilot study will be undertaken to evaluate this tool on clinical efficacy for safe SpTR, with patients being longitudinally assessed for neurological deficits. At the end of this study, the extensive evaluations will position the tool to a point of readiness for FDA submission. The AIP will lead to an enhanced pre-operative planning tool to plan safe SpTR, complementing intra-operative functional mapping, fulfilling a crucial unmet clinical need. The extended resection that this tool will facilitate, will potentially lead to extended survival times and hence improve patient outcomes. Thus, this tool is expected to significantly impact the clinical management of brain cancer, by affecting surgical treatment.

NIH Research Projects · FY 2026 · 2023-08

As defects in fetal adrenal (FAd) development can result in life threatening primary adrenal insufficiency (PAI), understanding the cellular and gene regulatory mechanisms governing this process is essential. While much has been learned about FAd development in rodent models, species-specific developmental differences limit our understanding of this process in humans. As mechanistic evaluation in human embryos is untenable, we recently developed the first human induced pluripotent stem cells (iPSCs)-derived FAd organoid system that recapitulates normal functional development and steroidogenesis. Using this system, we will undertake the first perturbative and reverse genetic assessment of human FAd development, allowing us to elucidate the molecular mechanisms of human adrenocortical development, which has broad implications in providing essential insight into mechanisms driving PAI. Development of the human adrenal cortex starts with specification of the adrenal primordium (AP) from the coelomic epithelium (CE), followed by establishment of the definitive zone (DZ) with putative stem cell/progenitor potential, and the fetal zone (FZ) with steroidogenic potential. Our recent single cell RNA-seq analysis of the human FAd cortex supports FZ replenishment by the DZ and the observed expression of Wnt ligands/activators in the peripherally located capsule (Cap) is suggestive of niche function. To mechanistically assess human FAd development and understand genetic defects (e.g., NR5A1, WNT4 mutations) driving PAI, we utilized our FAd organoid system. While the transcription factor NR5A1 impacts early FAd cell fate and promotes steroidogenesis in a gene-dose dependent manner in mice, its role in human FAd appears more complex. Our preliminary studies show that induced NR5A1 null mutant organoids fail to differentiate into AP-like cells (APLCs), exhibit decreased survival, fail to upregulate the receptor for adrenocorticotropic hormone (ACTH), and that the steroidogenic potential of the remaining cells is limited. In wild-type FAd organoids, we find that formation of DZ-like cells (DZLCs) is enhanced by the removal of Wnt inhibitor/addition of Wnt agonist. We also find that WNT4 and RSPO3 are highly expressed in both human Cap in vivo and Cap-like cells (CapLCs) in FAd organoids. Intriguingly, we find that DZLC formation is enhanced by sonic hedgehog (SHH), a trophic factor for murine Cap, suggesting a potential SHH-driven niche function for the Cap that is further supported by the observed upregulation of SHH target genes in human Cap. Finally, we find that both FACS-sorted DZ and DZLCs readily differentiate into FZ-like cells (FZLCs) upon ACTH stimulation, supporting the ability of DZLCs to replenish FZLCs. However, as Wnt-target gene expression is maintained in the subcapsular DZ despite exposure to ACTH, it suggests that Cap-derived Wnt signaling antagonizes ACTH- mediated differentiation of DZLCs. Using our organoid system, we will test the central hypothesis that dose- sensitive NR5A1 activity promotes development of the AP and that Wnt signaling provided by SHH- dependent responses in Cap both induces DZ development and antagonizes ACTH-driven FZ formation.

NIH Research Projects · FY 2024 · 2023-08

Project summary There is an urgent need to develop new vaccination strategies that induce antibodies that target relevant epitopes on complex microbes. Moreover, approaches that allow control of the number and fine specificity repertoire of responding naïve or memory B cells would fuel new vaccine designs for a variety of mutating viruses. This project centers on the hypothesis that co-engagement of the B cell receptor (BCR) for antigen with the Notch2 receptor will amplify responses by naïve and memory B cells. This hypothesis builds on preliminary data showing that BCR+Notch2 co-stimulation increases both number of B cells that respond and the number of proliferative events experienced by these cells. To test our hypothesis, we will generate unique mosaic nanoparticles based on immunogenic and sub-immunogenic variants of the HIV envelop (Env) antigen. We will answer the following questions: 1) Does BCR+Notch2 co-engagement prime naïve clones to target native-like Env variants?, and 2) Does BCR+Notch2 co-engagement on memory B cells increase their contribution to GC and PC pools? These studies will provide unique and needed insights into the potential exploitation of BCR signaling thresholds in emerging vaccine design.

NIH Research Projects · FY 2025 · 2023-08

Research: of givers complex FLG persistent aside of represents sociocultural as mutations risk development Candidate: Rico became Clinical currently funded focus time Chiesa dermatologist will Dr. with relationship, Biostatistics, Genetics benefit Resource-based Center (SBDRC) at Penn. Career focus completion knowledge become Atopic dermatitis (AD) is a chronic, inflammatory disease characterized by episodes acute disease flares, resulting in significant morbidity and disease burden to patients, care and health systems. While the etiology of AD remains unknown, it is thought to involve a interplay between genetic, immune, and environmental mechanisms. Mutations in the gene result in skin barrier dysfunction and are associated with more severe and disease in AD. However, little is known about the genetics of AD in other populations from those o European ancestry. Latinx are an admixed population and have higher rates AD compared to non-Latinx whites. The study of admixed populations such as Latinx an opportunity to examine the individual contributions of ancestry, genetics, and the interactions with genetic variation to the pathogenesis of complex diseases such AD. The main purpose of this study is to examine the presence of FLG loss of function (LoF) and (2) to explore the relationship between ancestry, genetic variation, and disease in Latinx with AD. Data obtained from this study, would then serve to inform the of future research to help us improve our diagnostic and management strategies. Zelma C. Chiesa Fuxench, M.D., M.S.C.E. graduated from the University of Puerto School of Medicine (2008). She completed her residency training at said institution and board-certified in dermatology (2013). Dr. Chiesa completed her Master of Science i n Epidemiology (MSCE) in 2016 at the University of Pennsylvania (Penn) where she is an Assistant Professor of Dermatology. Dr. Chiesa's current goal i s to become a R01 independent investigator focused on the epidemiology and genetics of AD with a specific on Latinx. Environment: During the time of this award, Dr. Chiesa will devote 20% of her to caring for patients with AD and 80% to clinical research work. Dr. Margolis and Dr. currently have a long-standing mentor-mentee relationship. He is a well-renowned and dermatoepidemiologist, and an expert in the genetic epidemiology of AD and serve as Dr. Chiesa's primary mentor. Dr. Chiesa will also benefit from the mentorships of oel M. Gelfand M.D. M.S.C.E., a renowned expert in the field of dermatoepidemiology and whom Dr. Chiesa has collaborated extensively as part of an established mentor-mentee Dr. Nandita Mitra, Ph.D., (Professor of Biostatistics at the Department of Epidemiology and Informatics at PSOM) and Dr. Sarah Tishkoff (Professor of Penn Center for Global Genomics and Health Equity at Penn). Dr. Chiesa will also from the rich training environment at Penn including the Development: Dr. Chiesa will primarily her training on genetic epidemiology and use of statistical methods in genetics. With of the proposed research and career development plan, Dr. Chiesa will acquire the and skills necessary to achieve a strong foundation to an independent, patient-oriented clinical researcher. f J Skin Biology and Diseases

NIH Research Projects · FY 2025 · 2023-08

Project Summary ART initiation (ARTi) is a unique clinical juncture in which virus replication and host immune responses are in flux and treatment of a substantial component of people living with HIV (PLWH) is possible. In both preclinical studies and recent clinical trials, infusion of broadly neutralizing antibodies (bnAbs) at ARTi has shown exciting preliminary results, but key questions about the mechanisms of action, the requisite bnAb properties, and the extent of clinical impact remain. Further, it is unclear if these benefits can be extended to the >35 million PLWH currently on suppressive ART, by using bnAbs after treatment interruption and ART re-initiation. Several clinical trials are planned or ongoing, but the complexity of human research limit the ability to definitively elucidate key mechanisms. Our scientific premise is that our molecularly defined, mixed bnAb-sensitive and resistant, barcoded transmitted/founder (TF) SHIV/NHP model system is uniquely poised to determine the extent and durability of bnAb activity at ARTi/re-initiation and decipher the mechanistic role of neutralization potency and effector function on reservoir dynamics, durable immune responses, and virus control. Our group has generated a body of work demonstrating that TF SHIVs reproduce key features of HIV-1 immunopathogenesis. We have expanded the model to incorporate genetic barcoding and virus inocula containing defined mixtures of bnAb-sensitive and resistant viruses. In this system, each animal is infected with a precise ratio of TF SHIVs encoding wildtype (WT; bnAb-sensitive) and escape mutant (EM; bnAb resistant) viruses, which have similar replication kinetics but markedly different sensitivities to V3-glycan bnAbs. Because WT and EM viruses have unique barcodes, we can track bnAb-sensitive vs. resistant virus clonotypes over time and across tissues through high-throughput sequencing, allowing for statistically powerful within-animal comparisons, as well as comparisons across treatment arms. Here, we will leverage this novel NHP system to determine the effects of bnAbs at ARTi and re-initiation and dissect the roles of bnAbs’ neutralizing and effector functions through a coordinated NHP experiment comparing 4 treatment groups: (i) ART alone, (ii) ART + bnAb, (iii) ART + bnAb with disabled effector function, and (iv) ART + bnAb with enhanced effector function. We will then determine if similar effects can be seen with use of bnAbs at ART re-initiation in animals already on suppressive ART who underwent ATI. This strategy allows us to define bnAb’s clinical impact and test our overall hypothesis that both the neutralizing potency and effector function of bnAbs at ARTi are essential to activity on the reservoir, host immunity, and induction of virus control. Successful completion of this project would have substantial significance to the HIV cure field, as it rigorously tests promising roles for bnAbs in HIV cure strategies that could guide the clinical development of bnAbs in cure strategies.

NIH Research Projects · FY 2025 · 2023-08

The American Consortium of Early Liver Transplantation-Prospective Alcohol-associated liver disease Cohort Evaluation (ACCELERATE-PACE) study is a prospective longitudinal cohort of patients with severe alcohol-associated liver disease (ALD) evaluated for early liver transplantation (ELT). The cohort leverages the ACCELERATE consortium with 4-linked R01s and 5 additional recruitment sites in the South/Southeast, Mid-Atlantic, Midwest, and West, and will refine and identify best practices in the selection and management of patients with severe ALD considered for ELT across their continuum of care. ALD is now the most common indication for liver transplantation (LT) in the U.S. Historically, LT centers required at least 6 months of alcohol abstinence before LT referral and evaluation, though empiric evidence to support minimum sobriety periods was limited. ELT, defined as LT before 6 months of abstinence, is increasingly performed but with significant practice variability. There is no consensus on optimal ELT candidate selection, and selection criteria vary widely, contributing to disparities in access to lifesaving care. ELT is also controversial due to the potential for liver recompensation with abstinence, which would obviate the need for LT—accurate prediction of recompensation has the potential to increase organ utility and stewardship. Detailed evaluation of the efficacy of alcohol use disorder treatments and improved risk scores based on pre-LT psychosocial factors to predict return to alcohol use are needed to refine selection criteria, optimize post-LT care, and effectively treat AUD. Short- and intermediate-term survival after ELT is excellent, but the incidence and predictors of post-LT complications are poorly defined. To fill these key knowledge gaps, we will enroll and prospectively follow 770 ELT candidates and 270 ELT recipients for 3 years at 9 centers across the U.S. The proposed Aims will: (i) inform ELT selection criteria and investigate potential sources of bias in ELT evaluation and healthcare disparities in ELT access; (ii) develop risk prediction scores for LT-free survival and recompensation; (iii) identify effective treatments (medical, behavioral) for alcohol use disorder among patients with severe ALD and post-ELT; (iv) evaluate clinical outcomes among ELT candidates and recipients, including mortality, transplantation, post-LT complications (e.g. cancer, cardiovascular events, graft rejection/failure), and quality of life. A comprehensive data repository will include sociodemographic, clinical, geospatial, psychosocial, behavioral, and patient-reported outcome variables. LT documents, checklists, recordings of selection meetings, direct observations of LT procedures, and clinician interviews will identify best practices and pitfalls in candidate selection. A biorepository of blood, urine, explant/donor tissue, pre- and post-LT liver tissue, peripheral blood mononuclear cells, and cross-sectional radiologic imaging will inform future ancillary studies.

NIH Research Projects · FY 2025 · 2023-08

Project Summary Multiple sclerosis (MS) is a neuroinflammatory autoimmune disease of the CNS that results in demyelination, axonal injury, and neuronal loss. Abnormal immune responses involving the trafficking of peripherally activated immune cells into the CNS are major drivers of inflammatory disease activity in relapsing multiple sclerosis, as underscored by the success of immune-targeting therapies. By contrast, the biology of non-relapsing progressive disease is thought to involve CNS-compartmentalized inflammation and degenerative disease mechanisms, which remains more refractory to therapy, due in part to the complexity of gene regulatory network coupled with cell-type-specific mechanisms of MS lesion progression. What types or subtypes of cells are affected by this process and their spatial heterogeneity in the tissue context as well as how these cells impact the tissue environments remain poorly understood, which precludes the development of strategies to target these cells to improve healthspan/lifespan or harnessing these cells or secreted factors to promote tissue remodeling and repair, highlighting a pressing need for tools to map cells and the surround environments in the tissues lesion and generate biomarkers to define spatial and phenotypic heterogeneity. This project aims to develop novel molecular barcoding scheme and downstream biochemistry in combination with novel microdevices for spatial multi-omics that allows simultaneous profiling of multi epigenomic modalities, whole transcriptome, and a panel of proteins at tissue scale and cellular level in a spatially resolved manner. We will apply the spatial multi-omics to map human brain tissue dissected from the edge of demyelinated white matter MS lesions at different stages of inflammation as well as the demyelinated lesion core, the white matter periplaque and normal white matter from neurologically healthy brains. Spatial omics data will be integrated with single-cell data to identify signatures of different affected cells and perform the tissue neighborhood analysis to define the cellular composition and molecular signatures in MS lesions. The expected outcomes and the major contributions include: (1) Fundamental knowledge on diverse cell types and their epigenomic, transcriptional and phenotypic (protein) characteristics in the context of 3D tissue organization in the MS brain lesions and (2) Offer the possibility of testing new therapeutic approaches for MS that are not targeted by currently approved treatments. The resulting data will lead to a better understanding of the relationship between tissue organization, function, and gene regulatory networks in MS.

NIH Research Projects · FY 2026 · 2023-08

Project Summary/Abstract Depression is the leading cause of disability worldwide and new treatments are needed. This therapeutic challenge stems from the fact that depression involves deficits distributed among neuronal subtypes and multiple brain regions. A growing body of clinical research demonstrates that a single dose of N-methyl-D-aspartate receptor , induce rapid and durable improvements in depressive symptoms persisting for days to weeks in patients refractory to conventional antidepressant therapies. Despite nitrous oxide being the oldest and one of the safest anesthetics in current (NMDA-R) antagonists, inhaled nitrous oxide or intravenous ketamine clinical practice, our understanding of how nitrous oxide modulates neuronal activity and circuit function to produce its clinical effects is extraordinarily limited. While it has been hypothesized that the therapeutic effect of NMDA-R antagonists is related to their ability to induce plasticity, the mechanisms that initiate and sustain this plasticity remain unclear. By combining in vivo imaging of synaptic structure, functional calcium imaging, electrophysiology, and behavioral recordings, this proposal will test an innovative hypothesis that changes in neuronal activity imposed by nitrous oxide acutely, automatically give rise to sustained plasticity through activity-dependent mechanisms. This hypothesis is based on extensive work on activity- dependent plasticity in the neocortex and on our own novel preliminary results showing that nitrous oxide specifically and directly activates layer 5 pyramidal neurons, which mediate cortico-cortical and cortico- subcortical connectivity, through a novel mechanism. All animal models and techniques have been established in the studies that generated the preliminary results, making the proposal highly achievable. In line with the mission of the NIGMS, the immediate goal of this proposal is to lay the foundation for advances in the treatment of depression by understanding the mechanisms of action of nitrous oxide in cortical circuits. Such an understanding may explain how acute pharmacologic interventions can lead to sustained therapeutic benefits in the setting of depression and potentially other neuropsychiatric diseases.

NIH Research Projects · FY 2024 · 2023-08

Opioid use disorder (OUD) is an ongoing public health crisis in the United States. Despite the effectiveness of current medications to treat OUD, there is still a high rate of relapse following detoxification. Thus, there is a critical need for a better understanding of the neurobiological mechanisms contributing to opioid taking and seeking that could lead to novel targets for future OUD treatments. An emerging literature studying the effects of opioids on human and rodent neural transcriptomes has identified alterations in gene expression in multiple brain regions. However, these studies are limited by bulk transcriptomic approaches, which prevent assignment of expression changes to specific neuronal or glial cell types. Single nucleus RNA sequencing (snRNAseq) approaches have the potential to address this knowledge gap and identify cell type-specific transcriptomic alterations that will advance our understanding of the pathophysiology of OUD. A recent snRNAseq study characterized the effects of an acute experimenter-delivered morphine injection on cell type-specific transcriptomes in the mouse nucleus accumbens (NAc), a brain region that plays a critical role in drug-taking and -seeking behaviors. Our preliminary studies extended these results by using snRNAseq to perform differential gene expression analysis in distinct NAc cell populations and subtypes following acute morphine injection or repeated morphine self-administration. Our exciting results reveal, for the first time, novel cell type- specific transcriptomic changes in the NAc that are associated with the escalation of voluntary opioid taking. A logical next step for the field is to determine the effects of sex on single nuclei transcriptomes as well as profile longitudinal changes in gene expression throughout subsequent abstinence periods. Therefore, one goal of this proposal is to characterize the sex- and cell type-specific effects of acute morphine exposure and volitional morphine taking in the rat NAc (Aim 1). One cohort of female Sprague-Dawley rats will receive an intraperitoneal injection of morphine or saline. A second cohort of female rats will be paired with yoked saline controls and allowed to self-administer morphine for 10 days, a period in which escalation of opioid taking develops. snRNAseq will then be used to identify sex-specific and sex-independent cell type-specific differentially expressed genes in the NAc. To comprehensively profile changes in gene expression throughout abstinence following morphine self-administration, snRNAseq will be used to analyze sex- and cell type-specific transcriptomic changes following acute (1 day) and prolonged (10 days) of opioid abstinence. Differentially expressed genes from each aim will be validated by fluorescent in situ hybridization (FISH) and RNAscope. This proposal will advance preclinical OUD research by using a snRNAseq approach to identify sex- and cell type- specific NAc transcriptome alterations associated with opioid taking and withdrawal thereby identifying novel therapeutic targets that will help guide the development of new medications to treat OUD.

NIH Research Projects · FY 2025 · 2023-08

The NIH Implementing a Maternal Health and PRegnancy Outcomes Vision for Everyone (IMPROVE) initiative aims to reduce maternal mortality and improve pregnancy, perinatal and postpartum care with a focus on community engagement. The University of Pennsylvania (Penn) proposes creating the Achieving Maternal Empowerment and Transforming Health through Implementation Science and Training (AMETHIST@Penn) Implementation Science Hub to support IMPROVE Maternal Health Research Centers of Excellence (CoEs) and other IMPROVE investigators, and to coordinate with NIH Staff Scientists, Project Staff, and the Data Innovation and Coordinating Hub. AMETHIST@Penn will be led by MPIs Meghan Lane Fall, MD, MSHP, and Rebecca Hamm, MD, MSCE, who bring complementary and necessary strengths in implementation science (IS) and maternal health. Dr. Lane-Fall is Executive Director of the Penn Implementation Science Center at the Leonard Davis Institute of Health Economics (PISCE@LDI). She is an internationally recognized acute care IS expert with an independent IS research program. She has 8 years of experience as a collaborator and consultant advising investigators on: selection of IS theories, models, and frameworks; implementation mapping and strategy selection; implementation outcome selection; data integration across multiple types and sources; and advanced analytic procedures to elucidate change mechanisms. Dr. Hamm is a maternal fetal medicine specialist who has established herself nationally as a researcher at the crossroads of obstetrics and IS. Together they will orchestrate the work of AMETHIST@Penn, leading an interdisciplinary, interprofessional group of more than 45 scientists and community partners. The AMETHIST@Penn Hub will comprise five Cores, each with two Co-Leads and 2-4 Core Investigators: 1) Administration, Leadership, and Evaluation; 2) Community and Stakeholder Engagement (with an Engagement Enhancement Collaborative designed to aid sites in accountability around community engagement); 3) Design, Methods, and Analysis (including a Consultant Panel with Methods, Community Engagement, and Clinical subpanels to meet the anticipated consultation and technical assistance needs of the CoEs); 4) Policy, Dissemination, and Communication (with skills building in messaging and media training); and 5) Training and Capacity Building (featuring an innovative academic- community partner training program and a comprehensive mentored research experience for both new and established investigators). In addition to establishing and administering the Hub, a research aim includes prospective mixed methods research on IS investigators, community partners, and institutions to characterize predictors of project-based implementation success and multilevel barriers to and facilitators of the conduct of maternal health IS research. The AMETHIST@Penn hub will advance public health and maternal health by facilitating rigorous implementation research that centers the needs and perspectives of patients, communities, and populations.

NIH Research Projects · FY 2024 · 2023-08

PROJECT SUMMARY/ABSTRACT The prevalence of opioid use disorder (OUD) among women giving birth in US hospitals increased by 131% from 2010-2017. This corresponds directly to the 82% increase in neonatal opioid withdrawal syndrome (NOWS) diagnoses in the same period, up to 7/1000 hospital births.1,2 Despite the need for maternal OUD treatment, referrals from postpartum units and neonatal intensive care units (NICU) to OUD treatment are low and postpartum treatment dropout is high among those who were treated prenatally.3 Infants born to women with OUD are often sent to NICUs after birth to manage opioid withdrawal, with their mothers at their bedside during their stay. An infant's NICU stay therefore represents a critical period in which to diagnose maternal OUD, initiate treatment, and refer for ongoing care in a setting that prioritizes and preserves maternal-infant dyadic bonding. Despite recommendations to integrate maternal mental health screenings and referrals in pediatric settings and a global move towards integrated mental health and substance abuse treatment in adult healthcare settings, limited information exists on how to close the gap in maternal mental health and substance abuse treatment in pediatric settings. The major challenge we aim to address is the need for adult treatment to occur in a pediatric treatment setting where structures do not yet exist to make this feasible. The goals of this study are 1) to systematically identify the primary facilitators and barriers to implementing bedside OUD treatment in the NICU through in-depth interviews with mothers, NICU providers, and other stakeholders; 2) to use steps from implementation mapping to develop a suite of implementation strategies to collocate buprenorphine induction for postpartum women with OUD in the NICU based on feedback from Aim 1; and 3) to refine, test, and examine the acceptability and feasibility of applying the adapted model in a pilot case series in two NICU sites for use in larger trials. This process will allow us to deliver a treatment model and set of implementation strategies for providing evidence-based OUD treatment to postpartum mothers in NICUs that will enhance its accessibility and sustainability. Future goals include testing the intervention and implementation strategies in a subsequent hybrid effectiveness- implementation study to estimate the extent to which they lead to improved implementation, sustained treatment, and outcomes for new mothers with OUD.

NIH Research Projects · FY 2025 · 2023-08

PROJECT SUMMARY Opioid use disorder (OUD) involving fentanyl is a major public health problem. OUD treatment with buprenorphine reduces all-cause mortality and drug-related morbidity and can be started by licensed prescribers in any outpatient or inpatient setting. For individuals using fentanyl, the process of starting buprenorphine is increasingly complicated by precipitated withdrawal. Withdrawal during initiation of buprenorphine deters some individuals from starting treatment and has been associated with treatment drop- out and relapse among those who do start. The goal of this project is to test the preliminary efficacy, safety, feasibility, and acceptability of a novel approach to initiating buprenorphine treatment for OUD that can be used in outpatient setting without requiring or precipitating opioid withdrawal. We will recruit 60 subjects with untreated OUD and recent fentanyl use through the University of Pennsylvania's Center for the Studies of Addiction. Subjects will be randomized to one of two arms: standard initiation, in which subjects will start buprenorphine after >8 hours of abstinence once they develop moderate opioid withdrawal with Clinical Opiate Withdrawal Scale (COWS) at least 11; or a novel low-dose (“micro-dose”) approach, which is started with COWS<4 and where buprenorphine doses are escalated over 10 hours, without ongoing use of full-agonist opioids. We will compare success rates with each approach, with “success” defined as reaching a total-daily dose of 8 mg buprenorphine without an increase of >6 in COWS from baseline and without early termination for any reason. Findings from this study will be used to support an R01 application to test this novel approach to initiating buprenorphine in real-world, outpatient settings with a larger sample of individuals with untreated OUD. This methodology has broad applications for increasing access to office-based treatment for OUD.

NIH Research Projects · FY 2024 · 2023-08

Alzheimer’s disease and related dementias (AD/ADRD) are devastating diseases to those diagnosed and come with a high cost to society. Reliable early identification and improved intervention is vital to treat patients before neuronal loss is irreversible. Autophagy is strongly implicated in the progression of AD, and thus has been an attractive therapeutic target for many years, but to reach a successful autophagy-targeting drug requires better understanding of the molecular consequences of disrupted autophagy. I propose to investigate how disrupted autophagy contributes to the chronic inflammation and plaque formation associated with the progression of AD. Using two of the most common AD risk variants as models to disrupt specific aspects of autophagy, I will investigate how the misregulation of mitochondrial DNA (mtDNA) and amyloid precursor protein (APP), autophagy cargos I identified in my postdoc, contribute to neuroinflammation, synapse loss, and plaque deposition observed in AD. AD initiation and progression involves multiple cell types, therefore I will use innovative iPSC models and cutting-edge techniques to model and manipulate complex interactions between neurons and microglia in a simplified system. In the K99 phase of this award, I will confirm that the ApoE4 AD risk variant disrupts mitochondria-endoplasmic reticulum contacts, which I predict will impair the clearance of mtDNA. I expect accumulation of mtDNA will sensitize neurons to release inflammatory factors, resulting in sustained microglia activation, release of complement and synapse loss. In the R00 phase, I will apply similar approaches mastered in the K99 phase to investigate the contribution of autophagy to plaque deposition. First, based off preliminary data, I will determine whether APP is an autophagy cargo in neurons or microglia, and whether it is normally transferred between the cell types. Second, as dysregulated Tau is a major disruptor of microtubules and organelle trafficking, I will investigate the sensitivity of TauR317W neurons to disruptions to autophagosome trafficking. I have found in my postdoctoral work that impaired autophagosome trafficking decreases degradation and increases secretion of autophagy cargo, thus I expect TauR317W sensitizes neurons to increase secretion and transfer of APP to microglia. I will then investigate the role of microglial autophagy to prevent plaque formation, and how this may be perturbed by the accumulation of Tau aggregrates and neurofibrillary tangles in TauR317W microglia. Completion of the independent aims will highlight the multifaceted role of autophagy in disease progression, identify specific molecular consequences of disrupted autophagy, and ultimately help to identify novel biomarkers and therapeutic targets for AD/ADRD.

NIH Research Projects · FY 2025 · 2023-08

Project Summary Aspirin is currently recommended during pregnancy to help prevent preeclampsia, and also shows promise for protecting against pregnancy loss. Preeclampsia is a dangerous complication of pregnancy that puts both the pregnant person and their baby at risk of serious morbidity and death, and pregnancy loss is the most common complication of pregnancy estimated to affect 20-30% of all conceptions. Current clinical guidelines from the U.S. Preventive Services Task Force (USPSTF) recommend the use of daily “baby” aspirin (81 mg), begun at 12 weeks' gestation, to prevent preeclampsia. However, data suggest that even earlier initiation of aspirin and at higher doses may result in greater protection. Furthermore, initiating aspirin immediately upon pregnancy recognition may maximize effects on preeclampsia while also providing the added opportunity to prevent pregnancy loss. Therefore, we propose to conduct a randomized trial to investigate whether daily “double low- dose” aspirin therapy (162 mg per day) initiated at the time of first positive pregnancy test (up to 6 weeks' gestation) may help prevent both outcomes. Clinical recruitment of participants through the follow-up of all initial pregnancy tests throughout the Penn Medicine system will ensure feasibility for early detection of pregnancy, enabling pregnancy loss monitoring and initiation of the trial intervention. The findings from this study will provide essential, high-quality evidence in a diverse sample of U.S. pregnant people to inform potential changes to clinical guidelines for aspirin use in pregnancy. We will (Aim 1) determine the effect of aspirin treatment (dose 162 mg) initiated upon positive pregnancy test (up to 6 weeks' gestation) compared to standard of care (dose 81 mg initiated at 12 weeks' gestation) on preeclampsia in a diverse population; (Aim 2) determine the effect of aspirin treatment (dose 162 mg) initiated upon positive pregnancy test (up to 6 weeks' gestation) compared to standard of care (dose 81 mg initiated at 12 weeks' gestation) on pregnancy loss in a diverse population; and (Aim 3) identify phenotypes who may benefit most from “double low-dose” aspirin (162 mg) begun soon after conception, using machine learning, to inform more targeted therapy for optimal overall pregnancy outcomes. As a safe, affordable intervention, these findings will have immediate translational value to improve reproductive outcomes in pregnant patients, including minorities such as Black women vastly underrepresented in current data.

NIH Research Projects · FY 2024 · 2023-08

PROJECT SUMMARY Multiple myeloma (MM) accounts for ~23% of all hematologic malignancies with a 2.1% of cancer-related deaths in the United States in 2022. Despite tremendous efforts to develop effective therapies, MM remains largely incurable, and virtually all patients develop resistance to current therapies. Thus, there is an urgent clinical need for innovative and improved MM therapeutics. It has been demonstrated that bone marrow endothelium is critical to MM cell homing, progression, survival, and drug resistance. Specifically, cyclophilin A and E-selectin, a homing factor and adhesion receptor, respectively, expressed by bone marrow endothelial cells, are critical to MM survival. Thus, inhibition of cyclophilin A and E-selectin provides a potential therapeutic strategy to abolish MM dissemination and resistance. However, direct- and specific-inhibition of cyclophilin A and E-selectin by small molecules has been elusive. Thus, cyclophilin A and E-selectin are promising candidates for combination RNA interference (RNAi) therapy, which inhibits traditionally undruggable targets by directly reducing their messenger RNA (mRNA) expression. The challenge of utilizing small-interfering RNA (siRNA) is the need for safe and effective delivery methods, as siRNA degrades in the bloodstream and does not readily cross membranes. During my predoctoral studies, I have engineered a library of polymer-lipid hybrid biomaterials, that in combination with polyethylene glycol (PEG)-lipid conjugates and siRNA, assembled into nanoparticles (NPs) via microfluidic mixing. Through high-throughput in vivo screening, I identified a NP formulation with potent gene silencing in bone marrow endothelial cells in vivo. This formulation was used to encapsulate cyclophilin A siRNA, and showed inhibition of MM progression in vivo, and sensitized MM cells to the proteasome inhibitor bortezomib, a current therapeutic modality to treat MM. During the F99 phase, I will improve our NP design by incorporating bone marrow endothelial-targeting ligands on the NP’s surface to enhance their specificity to bone marrow endothelium, minimizing off-target effects. I will use our targeted NP to co-encapsulate cyclophilin A and E- selectin siRNA sequences, and evaluate their inhibition in vitro through adhesion and transendothelial migration assays, to determine the invasive abilities of MM cells. Further, I will test our co-delivery siRNA nanotechnology through a survival study in a validated mouse xenograft model of MM and quantify its effects either alone or in combination with bortezomib. This technology is expected to provide with a broadly enabling platform to target other bone marrow-homing cancers. For the K00 phase, I will identify a renowned cancer biology laboratory to study cell-cell interactions in the bone marrow immune microenvironment utilizing high-dimensional single-cell approaches and tissue-engineered models, with the aim to determine mechanisms that drive cancer progression and drug resistance. Completion of this project will successfully prepare me to launch an NIH-funded research laboratory that focuses on drug delivery targeting the tumor microenvironment as means of cancer therapy.

NIH Research Projects · FY 2024 · 2023-08

PROJECT SUMMARY In response to hemodynamic demands faced by the heart, cardiomyocytes adapt and remodel primarily through changes in myocyte thickness or length. In conditions like cardiovascular exercise, changes in physiological hypertrophy can aid in improving cardiac function. However, in conditions of prolonged elevated blood pressure, heart rate, and excessive weight gain, pathological hypertrophy occurs and ultimately can contribute to heart failure. Changes in cardiomyocyte growth and atrophy that subsequently impact physiological and pathological cardiac remodeling are dictated by changes in protein synthesis. And while there is significant progress in understanding the transcriptional and translational changes that mediate maladaptive cardiac remodeling, little is known about the changes in the most abundant noncoding RNAs that link these two processes: transfer RNAs (tRNA). Outside of the notion that Polymerase III activity, which is required for tRNA transcription, increases in response to cardiac pressure overload, almost completely nothing is known about how alterations in tRNA transcription, transport, and localization impact sites of protein synthesis and, ultimately, cardiac remodeling. Considering that heart failure and the preceding changes in myocyte size contribute to the majority of deaths in the United States, there is an unmet need for new therapies that target malignant protein synthesis in pathological cardiac hypertrophy. Hence, this need may be met by identifying new regulating transcription and translational control from a novel tRNA-centric view. In this proposal, we will examine the relationship between tRNA transcription, transport, and localization in cardiac homeostasis and hypertrophy. We have identified that tRNAs, which are believed to primarily rely on passive diffusion for appropriate localization, require the microtubule network for proper localization in the heart. We hypothesize that pathological and physiological hypertrophy induce different changes in tRNA transcription; but that both types of growth require motor protein-mediated active transport of tRNAs along the microtubule network to facilitate necessary increases in protein synthesis for cardiomyocyte growth. We will test our hypothesis by carrying out the following aims: (1) To determine the role of microtubules in tRNA trafficking and localization in the cardiomyocyte. (2) To characterize the localization and expression of mammalian tRNAs during cardiac remodeling. (3) To identify the proteins that facilitate tRNA trafficking in cardiomyocytes. This work will be carried out in the laboratory of Dr. Benjamin Prosser, an expert on microtubules, cardiomyocyte mRNA trafficking, and cardiac remodeling. Successful completion of this work will have the positive impact of defining how changes in tRNA transcription, localization, and trafficking impact cardiac homeostasis and hypertrophy and thus result in the potential new strategies that target heart failure through the regulation of maladaptive protein synthesis and compromised cardiac cell size.

NIH Research Projects · FY 2025 · 2023-08

Summary Mitochondria are multifaceted organelles that play vital roles in a myriad of cellular functions, including energy production, metabolism, calcium homeostasis, and cell death. It is generally accepted that a decline in mitochondria quality is a key contributor to mitochondrial dysfunction, aging, and represents a key point of convergence for several neurological disorders. Yet, precisely how dysfunctional mitochondria contribute to these conditions remains elusive. Mitochondria are thought to be constantly rejuvenated via collaborative processes of mitogenesis, fission-fusion, and multi-level quality-control mechanisms. Accordingly, the average half-life of mitochondrial proteins in the brain has been estimated at less than 3 weeks. Recently, I identified a discrete number of mitochondrial long-lived proteins (mt-LLPs) that last at least four months in mouse brain and heart. These long-lived mitochondrial proteins (mt-LLPs) include OxPhos complexes and several mitochondrial cristae associated proteins, which similarly to other architecturally stable and long-lived structures (i.e. nuclear pore complexes) are recognized for their highly defined and elaborate ultrastructure. Therefore, we hypothesized that the exceptional longevity of mt-LLPs could play an essential role in the long-term stabilization of the mitochondrial cristae in long-lived, post-mitotic cells. The goal of this research proposal is to delineate the localization of mt-LLPs within mitochondria in neurons, examine their temporal dynamics and integration with newly synthesized proteins, and investigate their potential contribution to mitochondrial fitness and long-term cristae stability. In Aim 1, using a combination of pulse-chase protein labeling methods, super-resolution fluorescent imaging and mass spectrometry I propose to (1) examine the spatio-temporal dynamics of mt-LLPs in axonal and somato-dendritic domains of primary neurons. In Aim 2, we propose to extend our analysis to include mitochondrial DNA (mtDNA) by investigating the coordination between mt-LLPs enrichment and mtDNA longevity neurons. In Aim 3, I will investigate the mechanism(s) involved in persistence of mt-ELLPs in neurons using genetic manipulations targeting mitochondrial cristae stability. Lastly, in Aim 4 we will begin the investigation into the coordination between nuclear and mitochondrial genome expression in neurons. In summary, insights from the proposed experiments will significantly advance our understanding of long-term of mitochondrial proteome homeostasis and genome integrity in neurons, which could provide with new molecular targets for modulating the mitochondrial network dynamics in the processes of neurodegeneration.

NIH Research Projects · FY 2025 · 2023-08

The proposed projects are a comprehensive effort to rigorously investigate the contribution of phenology - or seasonal activity - to pathogen transmission dynamics using 3 widespread zoonotic pathogens carried by black-legged ticks. Variation in seasonal activity patterns and life-history events, which are well documented in species inhabiting temperate regions, can result in dramatic population dynamics including population extinctions or explosions. Phenological variation across time and geography also alters the frequency and strength of inter-species interactions and thus opportunities for pathogen transmission, which likely has profound impacts on pathogen transmission dynamics and human disease risk. Despite the likely importance to public health, the consequences of phenological variation on disease transmission dynamics remain notably under-studied in many complex zoonotic disease systems. The tick-borne disease system in North America provides the ideal system to determine the impact of phenology on pathogen transmission dynamics both theoretically and empirically. We will build and evaluate solvable analytical models, computational models, and advanced statistical models, all of which explicitly incorporate tick seasonal activity, and validate these models with empirical data from natural field sites. The proposed projects will (1) determine the quantitative impact of different phenological scenarios on the transmission dynamics of, and thus disease risk from, 3 human pathogens, (2) empirically evaluate model predictions, and (3) identify specific phenological and environmental features that result in different transmission dynamic outcomes among pathogen species. We will develop 3 modeling frameworks to investigate the impact of phenology on transmission dynamics, identify and quantify phenological and environmental drivers of transmission dynamics in 3 important human pathogens, and empirically validate the outcomes of the theoretical and statistical models. Empirical validation of both the phenological drivers and model predictions is essential not only for public health management but also to identify mechanistic processes driving these patterns. The proposed research provides both theoretical and empirical frameworks to investigate the impact of phenology on transmission dynamics in the multitude of disease systems involving multiple host species or life-stages from insect-vectored plant pathogens to pathogens with multi-host life-cycles.

NIH Research Projects · FY 2024 · 2023-08

Project Summary/Abstract The development and function of tubular organs, including the lungs, intestines, and inner ear, depends in part on an apical extracellular matrix (aECM) that lines the organ interior. aECMs contain many components that are shared across organisms such as mucins, collagens, and zona pellucida domain (ZP) proteins. Matrix proteins are often arranged into distinct layers or spatial domains within an organ; however, these structures are often destroyed by fixation, limiting the ability to study how they are formed. This proposal uses the well characterized and highly tractable model of the developing C. elegans vulva to understand how aECM is specified and assembled. Due to its transparent body and the ease of fluorescently tagging proteins, C. elegans is an ideal system to study aECM in a live organism. Many of the aECM domains in the vulva are cell type specific, allowing us to directly connect the assembly of matrix proteins to the biology of the cells they coat. This study focuses on two ZP matrix proteins, LET-653 and NOAH-1, that localize to the matrix of specific vulva cells. Like ZP proteins in some other systems, such as the mammalian inner ear, the localization of LET-653 and NOAH-1 is not well explained by their transcription patterns. ZP proteins require protease cleavage to properly localize and must also bind surface factors at their destination site. The identity of these ZP protein partners is unknown. Aim 1 will dissect the relative contributions of each cell type to the expression, processing and export, and matrix localization of ZP proteins. It will also identify the transcription factors required for cell type specific matrix fate. Aim 2 will identify candidate ZP protein partners with differential expression between cell types by single nucleus RNA sequencing (snRNA-Seq) and test their requirement for ZP matrix assembly. Ultimately, these experiments will uncover a gene regulatory network that specifies apical matrix identity and identify ZP protein partners that could be applicable to many systems. This project is designed to provide training in epithelial and matrix biology, as well as relevant techniques in single cell transcriptomics, genetics, and microscopy. It is sponsored by Dr. Meera Sundaram, an expert in matrix biology and C. elegans vulva development, and co-sponsored by Dr. John Murray, an expert in gene regulatory networks and single cell technologies in C. elegans. The research will be conducted at the University of Pennsylvania, an institution with a collaborative and stimulating intellectual environment as well as first-rate facilities and resources to conduct biomedical research. The proposed training plan incorporates many professional development opportunities available through the university, sponsors, and C. elegans community. At the conclusion of the proposed research and professional activities, the applicant will be prepared for a career incorporating teaching undergraduates and running her own laboratory.

NIH Research Projects · FY 2026 · 2023-08

Dental implants have become an important routine component of dental practice with over five million fixtures placed annually in the United States and this number is expected to increase significantly in the future. The peri-implant soft tissue interface is less effective than natural teeth in resisting bacterial invasion, enhancing vulnerability to subsequent peri-implant disease. Peri-implant diseases are inflammatory conditions affecting the soft/hard tissues surrounding a functional dental implant. Plenty of experimental evidence indicates that the accumulation of dental plaque at the soft tissue-implant interface and the subsequent local inflammatory response seems to be key in the pathogenesis of peri-implant mucositis. Furthermore, in certain individuals, it will progress to peri-implantitis, resulting in alveolar bone loss and implant failure. The goal of this application is to create a novel dental implant construct that renders the implant-supported restoration antibiofilm while providing a tight gingival tissue-implant seal that serves as a barrier to bacterial invasion. This smart dental implant system is a battery-less system that converts biomechanical forces from human oral motions (e.g., chewing or tooth-brushing) into electrical energy and powers light-emitting diodes that enable in situ phototherapy. When used in combination with a long-lasting antibiofilm restorative surface, this self-powered precision phototherapy system circumvents problems with the use of conventional antimicrobials. Ongoing studies indicate that red and near-infrared light is effective in maintaining human gingival tissue cell viability in the face of mono- and multi-microbial challenges. Furthermore, the antibiofilm restorative surface almost completely inhibits bacterial colonization. Based on these exciting supporting data, we hypothesize that force- powering of piezoelectric crystals to produce red and near-infrared light combined with bacterial anti-adhesive restorations creates an anti-inflammatory, pro-healing environment that provides a robust soft-tissue seal and prevents the development of peri-implantitis. We anticipate that the creation of this next-generation anti- inflammatory, antibiofilm dental implant system would increase functionality and provide a new strategy to prevent and control peri-implant diseases, especially in populations at risk, and reduce the risk of implant failure.