Mayo Clinic Rochester

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT One in four women undergoing lumpectomy require a repeat surgery because of positive margins. One of the underlying challenges in breast tumor margin assessment is a lack of fast, sensitive and reliable intraoperative tools. Fluorescence-guided surgery (FGS) using activatable contrast agents (ACAs) is emerging as an intraoperative tool for margin assessment during lumpectomy. Current design approaches to ACAs used in the clinic have been limited to targeting extracellular or cell surface proteins and they rely on covalently attaching a fluorophore and a quencher to a peptide substrate. Protease cleavage of this substrate separates the fluorophore quencher pair thereby resulting in activation of fluorescence. A major disadvantage of this approach is that cleavage of the peptide substrate occurs indiscriminately in both normal and malignant tissues giving rise to false positive signals. Studies have shown higher false positive rates with protease-targeting ACAs than with standard- of-care pathology. One could potentially circumvent this disadvantage by targeting alternative breast cancer- specific proteins. However, in the absence of a suitable design approach for non-protease ACA targets, it is not currently feasible to target non-enzymatic molecules. Here, we propose a new design paradigm that could be applied to detect any intracellular ligand-binding protein. We demonstrate our new approach with the estrogen receptor (ER) – a nuclear hormone receptor – which is upregulated in >70% of invasive breast cancer cases and is expressed at 8-10 times higher levels in tumor cells compared to normal cells. In our approach to developing ER-targeting ACAs, we propose to use supramolecular assemblies constructed from G-quadruplexes as hosts and bifunctional estrogens as the fluorescently activated moiety. The latter consist of a G-Quadruplex-binding fluorophore linked to an ER-targeting ligand. When bound to the G-quadruplex, emission of the fluorophore is quenched by guanines. In the presence of ER, binding of the ligand moiety displaces the fluorophore from the G-quadruplex supramolecular assembly thereby turning the emission “on”. In aim 1, we propose to optimize and characterize a panel of G-quadruplex supramolecular assemblies targeting the ER. In aim 2, we propose to validate the efficacy of the G-quadruplex assemblies compared to free probes for the specific detection of ER in vitro and in vivo. Our new design approach has broad applicability to any cancer-specific protein regardless of cellular location. With successful completion of aims 1 and 2 we will achieve our overall objective of using G-quadruplex assemblies as fluorescent ACAs to detect ER+ breast tumors. The establishment of a new design paradigm in general and the availability of ER-targeting ACAs will diversify the pool of targeted biomarkers for FGS to enhance the sensitivity and specificity of intraoperative margin assessment and ultimately improve patient outcomes.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY The extent of resection and size of the functional liver remnant after major hepatectomy for liver cancer are major factors affecting patient outcome and risk of post-hepatectomy liver failure. Therefore, therapies that stimulate liver regeneration without increasing the risk of tumor growth are vital to improving outcomes. Preliminary work by our German collaborators identified the dual-specific MAP2 kinase MKK4 as a master regulator of hepatocytic regeneration. In collaboration, we have identified a novel compound, HRX215, a highly selective small molecule MKK4 inhibitor with favorable pharmacokinetics that induced striking liver regeneration after partial hepatectomy in murine and porcine models of acute liver damage. Based on these favorable preclinical data, we hypothesize that HRX215 can promote liver regeneration in human patients. In the proposed phase 2b clinical trial, we will examine the effects of HRX215 in human patients receiving major hepatectomy at a single center, Mayo Clinic, Rochester, MN. In Aim 1, eighty patients undergoing portal vein embolization before major hepatectomy for colorectal cancer liver metastases will be randomized equally into treatment or control groups. Treatment patients will be administered 250 mg (PO) HRX215 twice daily for 28 days, initiated 12 hours before portal vein embolization. Control patients will receive a placebo. All patients will undergo volumetric computed tomography (CT) scanning before and on day 7 and day 28 after portal vein embolization. We hypothesize that the functional liver remnant in treatment patients receiving HRX215 will experience greater volumetric gain (indicating faster regeneration), and allow resection earlier and in a higher percentage of patients compared to the control group. In Aim 2, 40 patients undergoing major hepatectomy for hilar cholangiocarcinoma will be randomized equally into control and treatment groups, with control patients receiving a placebo and treatment patients receiving 250 mg (PO) HRX215 twice daily for 28 days following surgery. Both groups of cholangiocarcinoma patients will receive standard care, with volumetric CT scanning performed before and on day 7 and day 28 after resection. We hypothesize that patients administered HRX215 will have faster volumetric gain and reduced incidence of post-hepatectomy liver failure compared to control patients. Finally, we will use blood and tissue samples to measure intrahepatic neutrophil accumulation, quantify circulating myeloperoxidase and inflammatory factors, and perform spatial transcriptomics on the regenerating liver. These data will be used to determine the predictive value of novel blood and intrahepatic tissue markers in determining outcomes after surgery and estimating the risk of PHLF. We hypothesize that HRX215 will reduce post-resection inflammation and this response will correlate with better clinical outcomes. We believe these human trials could lead to new treatments for patients receiving major hepatectomy and identify a novel drug target for enhancement of liver regeneration.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT Deep Learning Reconstruction and Noise Reduction (DLR) algorithms are increasingly used in CT clinical practice, with substantial potential to reduce radiation dose and improve image quality. However, their advancement is hindered by the lack of accurate diagnostic performance evaluation tools and limited public CT projection data for training. These challenges create significant hurdles for developers to optimize their algorithms and prove dose reductions, and for regulatory bodies and clinical users to validate these claims, raising crucial safety concerns for millions of CT patients. With prior funding support from the NIBIB, we created a CT patient projection data library in an open, vendor-neutral format, DICOM-CTPD. This resource has been globally utilized by researchers to develop, train, validate, and test various DLR algorithms, including those mark the beginning of AI applications in CT imaging. Despite its great success, this library is constrained by its static dose levels, disease conditions, and data from older scanner models. These limitations highlight the pressing need for a new, dynamic patient projection data library with advanced diagnostic performance evaluation tools. The objective of this project is to develop a dynamic, customizable, vendor-neutral patient projection data library and software platform (DICOM- CTPD-VIT). This platform will be capable of generating a diverse range of conditions in radiation dose, reconstruction parameters, and lesion characteristics, and will facilitate automated image quality and diagnostic performance evaluation. The project comprises three specific aims: Aim 1: Develop a dynamic and vendor-neutral patient projection-data library with an integrated virtual- imaging-trial (VIT) software platform (DICOM-CTPD-VIT). Aim 2: Develop and validate a patient-data-based VIT evaluation framework for DLR methods. Aim 3: Develop and validate VIT-based approaches to quantifying hallucinations in DLR methods. The innovation and significance of this project lie in its dynamic, vendor-neutral patient projection data library, integrated with a VIT, and a patient-data-based evaluation framework. These resources allow for creation of varied imaging conditions, tailored to the needs of clinical users and researchers. It provides pathways for both algorithm development and clinical evaluation, addressing the diverse needs within the CT field to facilitate effective development and safe implementation of DLR methods in clinical practice. Moreover, the platform has the potential to generate an unlimited number of cases with ground truth, useful for training and evaluating other AI-based diagnostic tools beyond DLR.

NSF Awards · FY 2025 · 2025-08

This research will investigate membrane protein evolution with the long-term goal of designing artificial cells with tailored functions. The specific objective of this project is to determine how mutations in membrane protein transporters impact dynamics, conformational equilibria, and the direction of transport, which will provide insight into the biogenesis of membrane proteins. This research will involve and train students and postdoctoral associates in biochemistry and biophysical chemistry. Professional training opportunities will be afforded to strengthen communication skills and to prepare trainees for future career paths. The project will also involve the development of a class for graduate students with the goal of teaching students advanced techniques in biochemistry and biophysical chemistry. The broader goal of this endeavor is to strike to a more equitable balance between lecture-based curricula and hands-on learning in graduate education. Many membrane protein transporters are comprised of a single polypeptide chain that contains inverted repeats within its structure. This research will investigate the underpinnings of membrane protein evolution by using mutations to derive a quantitative relationship between function and the free energy distinguishing conformational states required for function. The functional experiments will utilize a flow cytometry-based method capable of analyzing a library of mutations in a high throughput manner. To correlate the free energy with function, a quantitative method involving 19F NMR spectroscopy will be developed. Finally, theories about the evolution of ion-coupled transport mechanisms will be tested by using directed evolution experiments and bioinformatics. This project is supported by the Molecular Biophysics Cluster in the Division of Molecular and Cellular Sciences. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY ABSTRACT Cholangiocarcinoma (CCA) is a highly lethal malignancy of the liver and biliary tract, with incidence and mortality rates steadily increasing worldwide. Despite advancements in cytotoxic therapy, the 5-year survival rate remains below 10% due to high rates of tumor recurrence, drug resistance, and toxicity. This highlights an incomplete understanding of the mechanisms regulating CCA cell death. Ferroptosis, an iron-dependent and regulated mode of cell death driven by phospholipid peroxidation, has emerged as a promising alternative for inducing cell death in tumors resistant to conventional therapies. However, the therapeutic benefits of ferroptosis in CCA are poorly defined, and its induction in vivo faces challenges like poor selectivity, off-target effects, and acquired ferroptosis resistance. Resistance to cell death often results from inactivating mutations in tumor suppressors, such as BRCA1-associated protein 1 (BAP1), which is mutated in up to 25% of CCAs and associated with ferroptosis resistance in renal cell carcinoma. This proposal aims to elucidate the therapeutic benefits of ferroptosis in both sensitive and resistant CCA cells in vitro and in vivo. Preliminary data show that BAP1 inactivation or downregulation confers resistance to ferroptosis in CCA cells, while ferroptosis induced by glutathione peroxidase 4 (GPX4) inhibitors in mice bearing BAP1 wild-type CCA reduces tumor growth. Additionally, EpCAM- aptamer coated nanoparticles packaged with siRNA against GPX4 efficiently target CCA cells, demonstrating tumor enrichment and good tolerability. Our central hypothesis is that ferroptosis provides a novel therapeutic opportunity in both BAP1 wild-type and BAP1-mutant CCA cells, maximizing cytotoxic effects. To investigate this hypothesis, we have outlined two specific aims: 1) evaluate the therapeutic efficacy of EpCAM-aptamer coated nanoparticles with siRNA against GPX4 in inducing ferroptosis in BAP1 wild-type CCA; 2) identify genes mediating ferroptosis resistance in BAP1-mutant CCA cells using a genome-wide CRISPR/Cas9 loss-of-function screen. The proposed research will uncover new targets underlying ferroptosis resistance in BAP1-mutant CCA, revealing metabolic vulnerabilities for future therapeutic interventions. Our approach includes EpCAM-aptamer coated nanovesicles for selective targeting, CRISPR knockout libraries, and sophisticated cellular and animal models to examine ferroptosis and tumor characterization.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Healthy human cells contain 92 telomeres (2 arms per 46 chromosomes), each with unique lengths that shorten during cell division, and that protect the genome from enzymatic degradation. Average telomere length (TL) of all 92 telomeric alleles has clinical relevance as a biomarker for cellular aging and a broad variety of health-related conditions, including telomere biology disorders (TBD): inherited disorders that exhibiting idiopathic pulmonary and liver fibrosis, bone marrow failure and cancer predisposition. Telomeres also play an important regulatory role in gene expression through the telomere position effect (TPE). However, the clinical relevance of these individual 92 chromosome arm-specific TLs and their connection with TPE is still unclear. None of the existing methods to measure TL are at the same time accurate, scalable, suited for any tissue source, or able to provide length of individual telomeres. This includes Flow-cytometry Fluorescence In-Situ Hybridization (flowFISH), the only CAP/CLIA-approved test to measure TL. Recent advances in long-read sequencing technologies have allowed our group to develop Telogator, a novel bioinformatics approach that estimates TL for all 92 alleles, individually, from this data. We propose to use this approach in combination with multiomics analyses to test the main hypothesis that individual chromosome arm-specific TL are clinically relevant in the diagnosis and understanding of TBD, and that individual shortened telomeres can trigger transcriptomic and epigenomic changes involved in the pathogenesis of the disease. Through the Mayo Clinic TBD Specialty Clinic, we have managed over 150 patients with manifestations of TBD including flowFISH and genetic germline testing. These individuals are clinically and molecularly annotated, they have already given consent for research through our IRB-approved protocol and PBMC samples are readily available. We will use this unique resource to test our hypothesis by completing the following aims: In Aim 1 we will define the reference TL values at the chromosome arm level across the lifespan of healthy individuals using our novel sequencing-based bioinformatic tool (Telogator) on 30 individuals distributed in age decades from their 20s to their 70s. We will then compare these reference values with the length of individual telomeres from TBD patients (n=30) calculated using the same approach. In Aim 2 we will characterize the transcriptomic (RNAseq) and epigenomic (ChIP-seq) changes in correlation to the TL of each individual chromosome arm to describe genes regulation through TPE in the context of TBD. This project will explore the conceptually innovative idea that TL of individual chromosome arms has clinical significance and can exert characteristic epigenomic and transcriptomic changes in human cells. Our research will translate into better diagnostic tools for TBD and pave the way for future research in the pathogenesis of the disease. Understanding the gene regulation related to chromosome-specific TL will facilitate the study of other telomere-related processes like aging and cancer.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Chronic villitis of unknown etiology (VUE) is commonly diagnosed in the placenta and key features during histologic evaluation are the infiltration of T lymphocytes into the villous parenchyma and corresponding trophoblast necrosis. VUE is seen in the context of adverse fetal outcomes (i.e., fetal growth restriction, NICU admission, neurodevelopmental delays) and seemingly normal outcomes. As our understanding of the cause of this inflammation is not well defined, it is difficult to understand the spectrum of fetal outcomes associated with a VUE diagnosis and develop useful clinical management strategies which could be employed during pregnancy to positively impact health. The current hypothesis for VUE is that it represents a breakdown of immunologic tolerance to the fetal allograft resulting in maternal T cell targeting of paternal (fetal) antigens in the placenta. We and others have previously shown that there are histologic and immunologic changes associated with villitis, specifically as it relates to human leukocyte antigen (HLA) expression; however, we still cannot predict or prevent VUE from harming a fetus, which is particularly significant for couples worried about recurrent villitis. Therefore, this study aims to provide stronger evidence to delineate the immunology of VUE based on severity and immune mediated rejection pathways. Our hypothesis is that VUE represents an immunogenic pathology resembling organ rejection and that certain maternal-infant HLA types lead to increased risk of disease and poor fetal outcomes. We will address this hypothesis in two Specific Aims: 1) evaluate the associations between maternal-infant haplotype and VUE (adverse fetal outcomes vs. normal outcomes); and 2) determine whether VUE placentae have activation and upregulation of organ rejection genes/pathways in the absence of infectious stimuli. By studying the underlying VUE pathogenesis associated with positive and negative fetal outcomes, we will be able to discover better therapeutic targets, create valuable risk stratification tools and even identify biomarkers of placental inflammation which could be used to manage VUE in utero and prevent the poor neonatal outcomes associated with this pathology.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Inflammatory bowel disease is characterized by remitting and relapsing bouts of intestinal inflammation and symptoms of abdominal pain, dysmotility, and autonomic dysfunction persist beyond the periods of active inflammation. Evidence suggests that neural pathways of the Gut-Brain Axis, particularly the sympathetic nervous system, are involved in pathophysiology, but until recently technical restrictions have limited the ability to fully understand the underlying neural circuit mechanisms. Therefore, the proposed training and research plans are focused on developing the Applicant’s expertise in sympathetic regulation of gastrointestinal function in health and disease. The training plan focuses on development of innovative technical skills and comprehensive career mentorship with the goal of providing the Applicant a complete set of skills necessary to develop into an independent scientist investigating the Gut-Brain Axis. The research plan complements this training by investigating how sensory-sympathetic neural pathways regulate the outcomes of colitis and recovery, specifically delineating the mechanisms used by two distinct circuits innervating the colon: (1) extrinsic primary afferent neuron (ExPAN)-spinal-sympathetic pathways and (2) intestinofugal afferent neuron (IFAN)-sympathetic pathways. This will be achieved through two Specific Aims utilizing innovative ex vivo imaging techniques with optogenetic reporters in intact sensory-sympathetic colon circuits, advanced in vivo chemogenetic approaches, and cutting-edge in vitro systems with patient-derived organoids and microfluidic devices. Aim 1 will define the changes in ExPAN- and IFAN-mediated activation of sympathetic neurons in the prevertebral ganglia (PVG) during active colon inflammation and after recovery. Aim 2 will determine the role of PVG sympathetic neurons for dysmotility and mucosal healing in colitis. The proposed research and training will significantly advance the applicant's expertise in cutting-edge techniques and technologies related to gut-brain signaling, while uncovering novel mechanisms that address a critical gap in our understanding of sympathetic regulation in colitis.

NIH Research Projects · FY 2025 · 2025-08

ABSTRACT Sporadic outbreaks of Ebola virus (EBOV) and related filoviruses pose a grave threat for worldwide health populations, especially those of Western African nations. The largest outbreak of EBOV in 2013 caused nearly 13,000 deaths and had case fatality rates of up to 90%. Despite its near half-century since its emergence in human populations, there is only one vaccine and only two recently approved therapeutic treatments for Ebola virus disease (EVD). Detailed molecular study of EBOV biology is necessary to rapidly advance antiviral development. The largest impediment to detailed molecular study of EBOV is the requirement for high containment facilities when handling infectious virus. As an alternative, many groups utilize the well-established life-cycle modeling of both minigenome and transcription and replication virus-like particles (trVLP) systems to assess molecular viral mechanisms under biosafety level-2 (BSL-2) conditions. To this end, we have generated a penta-cistronic minigenome (5XMG) construct that contains four of the viral open reading frames and a reporter gene. The minigenome system remains BSL-2 due to its inability to replicate unless in the presence of the replication “helper” proteins: NP, VP30, VP35, and L. While this is valuable as a safety mechanism, it inhibits the use of EBOV modeling in animals. It would be invaluable to have a murine model in which to study the dynamics of EBOV under BSL-2 conditions in vivo. The proposed study will generate two mouse models for studying EBOV dynamics using trVLPs by supplementing the necessary helper proteins in trans. We would achieve this through two specific aims: (Aim 1) In Vivo EBOV Life Cycle Modeling with trVLPs via EBOV Helper Protein Expression by mRNA-Lipid Nanoparticles (LNPs); (Aim 2) In Vivo EBOV Life Cycle Modeling with trVLPs Supported by EBOV Helper Protein Expression via Transgenesis. In this model, the proteins missing from our poly-cistronic minigenome (EBOV VP30, VP35, and L) would be expressed in mice via either mRNA encapsulated LNPs or as integrated, Cre-recombinase dependent mouse transgenes. Expression of the viral helper proteins will replicate our 5XMG to amplify and create additional trVLPs that would bud off, infecting additional cells in the animal’s tissues. Importantly, the proposed system would be a self-contained model for infection with minigenome-containing trVLPs but will not generate any infectious virus. These tools can have many applications including the assessment of antiviral host responses as well as being a platform for testing antiviral therapies like vaccines and small molecule inhibitors.

NIH Research Projects · FY 2026 · 2025-08

PROJECT DESCRIPTION/ABSTRACT Glioblastoma, IDH-wildtype (GBM) is the most common and most lethal primary brain cancer. Despite tremendous research efforts, the last FDA drug approval for newly diagnosed GBM was temozolomide in 2005, and these tumors invariably recur with a median survival of only 16 months. Thus, there is a tremendous unmet medical need to develop effective therapies for this disease. Antibody drug conjugates (ADCs) are a rapidly growing class of molecularly targeted therapeutics with impressive clinical activity in various solid malignancies. ADCs capitalize on tumor-specific monoclonal antibodies to deliver highly potent toxins into tumor cells, while limiting exposure to normal tissues. Given this potential for selective tumor therapy, we have been investigating the potential of ADCs in GBM for over five years. GBM express a number of tumor antigens for which ADCs have been developed including Epidermal Growth Factor Receptor (EGFR) and B7 family of immune-regulatory checkpoint protein (B7H3). In partnership with AbbVie, members of our team evaluated three EGFR-targeted ADCs in a panel of GBM patient-derived xenografts (PDXs). These studies demonstrated that i) most EGFR-amplified GBM grown as heterotopic tumors are profoundly sensitive to these ADCs, ii) restricted delivery across the blood-tumor barrier after systemic dosing limits the efficacy of the same ADCs in most orthotopic PDXs, iii) direct infusion of ADCs into brain tumors using convection enhanced delivery (CED) can be highly efficacious, and iv) neuronal toxicity of ADCs delivered into the brain is highly dependent on stability of the linker. These key findings provide the overall rationale for this R01 that is focused on engineering ADCs specifically designed to enhance the therapeutic window for treatment of GBM and other cancers within the central nervous system. Importantly, the work proposed is only possible with the collective expertise of our key team members: Dr. Kyoji Tsuchikama (MPI) is a medicinal chemist and leader in ADC engineering, Dr. Jann Sarkaria (MPI) is an expert in translational development of novel therapeutic strategies for GBM, and Dr. William Elmquist (Co-I) is a world-leader in the pharmacokinetics of drug distribution in the central nervous system. With this team, we will address key challenges in the development of ADCs as a therapy for GBM in three specific Aims. Aim 1: Develop novel linker chemistries to improve stability of ADCs in the brain Aim 2: Evaluate the potential to limit neuronal toxicity through optimized toxin selection Aim 3: Evaluate novel ADC therapeutic strategies to address tumor heterogeneity

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT Grade 4 astrocytomas are primary brain tumors with a median survival of 15-31 months, depending on the isocitrate dehydrogenase (IDH) mutation status. Despite maximal safe surgical resection and aggressive chemoradiation, these tumors inevitably recur fatally. Disease monitoring relies on magnetic resonance imaging (MRI), as assessed by the Response Assessment for Neuro-Oncology criteria (RANO 2.0), which is imperfectly sensitive to disease progression. Specifically, evaluation of disease progression can be hampered by pseudoprogression, or increased contrast-enhancement post-treatment without an actual change in tumor burden. Additionally, significant disease progression must occur before meeting the >25% increase in radiographic size to diagnose progression per current response assessment criteria, preventing early action when disease burden is more minimal. Limited tissue access post-resection underscores the critical need for non-invasive, highly sensitive, and specific monitoring tools. Our project addresses this need by developing a novel liquid biopsy-based assay using plasma circulating tumor DNA (ctDNA) to detect amplified genomic chromosomal junctions unique to each patient. These junctions, resulting from chromosomal rearrangements under selective pressure, are highly specific to the patient's tumor. Leveraging our team's prior experience with this assay in other cancer types, our data to date in patients with high-grade gliomas (HGGs) demonstrate excellent sensitivity and specificity for disease detection, including at baseline and through disease recurrence. In this project, we will perform the technical and clinical validation studies necessary to deploy the amplified junctions ctDNA plasma test as a clinical assay in patients with HGGs. In the UH2 phase, we will validate the technical aspects of our amplified junctions ctDNA assay, starting with low-coverage whole genome sequencing of tumor tissue to identify informative chromosomal junctions. These will be used to generate primers that can then be deployed to detect the abundance of junctions in plasma via PCR. Each step's accuracy, precision, sensitivity, specificity, and quality control measures will be thoroughly assessed, with specific go/no-go criteria required for transition to the clinical validation phase (UH3). The UH3 phase aims to establish the clinical utility of this approach, as defined by the clinical endpoint of detecting changes in disease burden, including response to cytoreduction or disease progression. We will assess changes in junction abundance relative to cytoreductive therapy, stable disease, and progression, distinguishing true increases in disease burden from pseudoprogression. This phase will also evaluate the utility of our assay to detect progression earlier than MRI. In summary, this project not only proposes a robust method for glioma monitoring but also integrates advanced genomic tools and personalized medicine approaches to significantly enhance response assessment accuracy in patients with HGGs, toward our goal of improving outcomes by enabling more timely therapeutic interventions.

NIH Research Projects · FY 2026 · 2025-08

Project Summary Anthracyclines are highly effective chemotherapeutics, but one of the main side effects limiting their use is cumulative dose-dependent cardiac toxicity, also known as Anthracycline-Induced Cardiotoxicity (AIC). Cardiac responses of patients to anthracyclines are highly variable, and the underlying genetic underpinnings remain largely obscure. Dr. Xu’s group has been developing zebrafish as an efficient vertebrate model for discovering genetic modifiers of AIC. Our recent screens identified fabp7 and gpnmb as two AIC modifiers, detailed studies of which suggested a novel epicardium-based aging-associated therapeutic strategy for AIC. In the meantime, Dr. Norton’s group has been carrying GWAS studies and exome sequencing of human AIC patients. A large number of clinically relevant candidate variants have been identified; however, precise genetic factors are often difficult to conclude. We reasoned that efficient zebrafish genetics could be used for experimentally testing candidate genes suggested from human GWAS studies. We have generated substantial preliminary data, prompting us to propose to demonstrate that modifier screens in zebrafish facilitate the discovery of novel susceptibility genes by testing candidate genes suggested from human GWAS studies and novel pathological events such as aging-associated epicardial remodeling. Our efforts can be divided into the following three specific aims. In Specific Aim 1, we propose to continue to study fabp7 and gpnmb, aiming to confirm their identity as therapeutic target genes for AIC. To obtain direct evidence supporting that their cardioprotective effects are related to cardiac aging, we will explore killifish, a vertebrate with a short lifespan, as a new animal model for studying cardiac aging. In Specific Aim 2, we propose to test the epicardial remodeling hypothesis by detailed expressional studies of fabp7 and gpnmb, followed by conditional genetic studies. In Specific Aim 3, we will discover additional aging-associated modifier genes via utilizing zebrafish genetics to experimentally test ~23 candidate genes suggested from GWAS studies of AIC patients. The deliverables of the proposal are: 1) we will establish epicardial remodeling as a previously unrecognized aging-associated pathological event in AIC. 2) fabp7 and gpnmb could be established as the first two epicardium-based target genes that can be harnessed for therapeutic benefits on AIC. 3) 5 additional AIC modifier genes, including about 3 with epicardial expression, could be identified. Our proposal will lead to the establishment of a fish-human platform that enables systematically discovery of AIC modifying genes.

NIH Research Projects · FY 2025 · 2025-08

In 2017, the All of Us (AoU) Research Program was established to realize the ambitious goals set out by the Precision Medicine Initiative proposal. The goal of the AoU Research Program is to enroll one million people living in the U.S. into a national cohort that broadly represents the U.S. population and to accelerate progress toward a new era of precision medicine on a larger scale than previously possible. Over the course of the past 7 years, the AoU Research Program has enrolled 848,783 participants with 608,145 providing biospecimens, resulting in 3,959,288 samples collected and 14,521,323 aliquots created and stored at the AoU Biobank housed within the Mayo Clinic Biorepositories Program. The Mayo Clinic Biorepositories Program is CAP accredited and has biobanking activities established at all three Mayo Clinic sites (Rochester, MN, Jacksonville, FL, and Phoenix, AZ). Mayo Clinic has been an integral member of the AoU team since its inception, supporting the AoU Research Program through input on program development and execution of all activities related to biospecimen collection, processing, storage, distribution, and management. The objective of this new proposal is to continue utilizing the full resources available through the Mayo Clinic Biorepositories Program to efficiently and cost effectively support the AoU Research Program in innovative ways with respect to AoU Biobanking activities, namely: biospecimen collection, kit assembly, biospecimen shipping, receipt, accessioning, tracking, processing, quality assurance, storage, and distribution. Mayo Clinic Laboratories in Rochester MN, the third largest reference laboratory in the United States, will continue to support AoU Biobank activities by enabling efficient specimen transportation and routing between AoU collection sites and the AoU Biobank at Mayo Clinic. While Mayo Clinic Rochester will continue to serve as the primary site for AoU Biobank activities, the biorepository group at Mayo Clinic Jacksonville will continue to provide support as the secondary site for sample storage, ensuring that a subset of the cohort is available as a backup. The specific aims of this proposal are to: 1) Maintain Existing Biospecimens and Sustain Biobank Operations; 2) Support Biobank Expansion Initiatives, Including Pediatric, Reassessment, and Ancillary Studies; and 3) Provide Ongoing Collaboration and Support for the AoU Research Program. Overall, the combined capabilities and facilities available through the Mayo Clinic Biorepositories Program and Mayo Clinic Laboratories provide an unparalleled ability to collect, accession, process, distribute, store, and manage virtually any type of biospecimen, while maintaining the highest standards for biospecimen quality. As demonstrated over the course of the past 7 years, we have the capacity and the proven capabilities to support all new initiatives on behalf of the AoU Research Program.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT Therapy for high-grade serous ovarian cancer (HGSOC) relies on DNA damaging agents, with a high percentage of cancer with cyclin E amplification or overexpression having de novo or acquired chemoresistance due in part to homologous recombination (HR) proficiency. We recently discovered that the anti-leukemic drug clofarabine (CLF) + olaparib combination aided in the nuclear translocation of cathepsin L (nCTSL) to induce DNA damage and sensitize OC cells to PARP inhibitors (Funding by DoDIIRA, OCRP venue.) Although our ongoing studies substantiate the role of nCTSL in the DNA damage response (DDR), the precise mechanism by which nCTSL induces DNA damage remains unclear. Our preliminary data has uncovered a potential mechanism that promotes homologous repair deficiency (HRD) in CCNE1 high tumors. CLF + ATR inhibitor (ATRi), AZD6738 induced an increase in the replication stress (RS) marker pRPA2S33 foci validated through immunofluorescence (IF) and cell fractionated western blot analysis. This coincided with the presence of CTSL in the nucleus. ATR knockdown (KD) or ATRi combined with CLF, synergistically decreases OC cell survival, observed in both CCNE1 high and low expressing cell lines and in ex vivo cultures of patient derived xenograft models in vitro. Significantly, CTSL KD confirms nCTSL's role in CLF-induced replication stress. This proposal aims to develop novel approaches to evaluate drug induced replication stress marker as a means of developing novel drug combinations in high grade serous ovarian cancer in vivo and mechanistically determine the how nCTSL attenuates CCNE levels. Continued development of novel combinations targeting replication stress is essential, particularly in patients with PARPi resistant/recurrent tumors. The goal of this proposal is to test our hypothesis that nCTSL mediated CCNE1 downregulation may sensitize CCNE1 driven tumors to PARPi and ATRi treatment and restore homologous repair deficiency in vivo. The project will 1) Demonstrate that the cytotoxicity of CLF + ATRi and CLF + PARPi saruparib is associated with nCTSL in CCNE1 high PDX models in vivo and in syngeneic mouse model using KPCA cell line CCNE1 overexpression. 2) Determine the mechanism by which nCTSL targets CCNE1. Continued development of novel combinations targeting replication stress is essential, particularly in patients with PARPi resistant/recurrent tumors. Considering the challenge of resistance to first-line therapy and PARP inhibitors, the combination of CLF with ATRi holds promise in overcoming or delaying drug resistance in recurrent in high grade serous ovarian cancer.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT Problem To Be Addressed The large number of surgical management options for BPH can be overwhelming for patients and providers alike. To better understand the factors that impact patient uptake of surgical treatment for BPH, it is important to understand specific patient preferences, including attitudes regarding cost. Specific Aims and Experimental Design Aim 1: Identify the peri-operative factors most important to patients undergoing BPH surgical intervention. Aim 2: Quantify patient priorities surrounding BPH surgical intervention. Aim 3: Assess the relative importance of cost for patients undergoing BPH surgical intervention. Beginning with an ongoing study utilizing validated questionnaires (EQ-5D-5L, OAB-5D, COST-FACIT, and patient preference ranking) provided to 150 patients undergoing BPH surgery, we will identify the factors (e.g. procedure location, indwelling urethral catheter duration, risk of urinary incontinence, and retreatment rate) most important to patients (Aim 1). These factors will inform the construction of a subsequent discrete choice experiment (DCE) survey. Patients will be asked to choose among multiple choice tasks that can quantify the trade-offs for positive and negative attributes for hypothetical BPH surgical interventions (e.g., lower risk of short-term incontinence but higher risk of need for retreatment). In addition to the factors identified on the initial survey study, a range of costs will be included to ascertain willingness-to-pay (WTP) thresholds for the various attributes prioritized by patients. The DCE will be prospectively administered online to 200 patients undergoing evaluation of BPH within the urology clinic. Each patient will value 10 to 15 choice tasks. Econometric analysis will be performed to identify preference weights, overall attribute importance, attribute equivalence for each factor included, and WTP estimates (Aims 2 & 3). These data will serve as the first quantifiable measure of preferences for patients undergoing BPH surgery. Findings from this study will serve as a foundation for subsequent patient and provider decision aid tools. Career Goals and Development Through work with a multidisciplinary mentorship team at Mayo Clinic, the candidate will gain knowledge and skills needed for survey-based research, qualitative analysis, DCE methodology, and shared decision-making tool creation for BPH and other benign urologic conditions.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY/ABSTRACT Despite a characteristic indolent course, a substantial subset of follicular lymphoma (FL) patients has an early relapse with a poor outcome. Thus far, efforts to identify factors that predict survival have been unsuccessful. We found that FL patients with a worse prognosis display an increased expression of IRF4. While IRF4 normally promotes terminal differentiation of B cells to plasma cells, its overexpression in FL B cells dysregulates immune signaling and induces an immunosuppressive microenvironment. Our preliminary data show that IRF4 controls the expression of multiple immune recognition molecules, suggesting its role in modulating the interaction between B and T cells. Additionally, silencing of IRF4 in vitro and in vivo led to memory B cell (MB) transcriptional reprogramming and cell differentiation. At the molecular level, there was an inverse correlation between IRF4 and the MB transcription factor (TF) BACH2, suggesting that their mutual antagonism may dictate cell fate. We hypothesize that IRF4 controls B cell fate in counterbalance with BACH2, and disruption of this mechanism promotes lymphomagenesis. To test our hypothesis, we propose two specific aims. In Aim 1 we will use new transgenic mouse models with overexpression and deletion of irf4 in germinal center B cells to investigate whether and how IRF4 counteracts BACH2 to define B cell fates by integrating single-cell transcriptional/protein (CITE-seq) analysis and DNA-binding (CUT&RUN), the latter to identify IRF4 and BACH2 target genes. In Aim 2 we will perform CITE-seq and CUT&RUN of malignant B cells from the transgenic mouse models in Aim 1 to elucidate whether the imbalance between IRF4 and BACH2 promotes lymphomagenesis. These findings will be validated using single cell RNA-seq of human FL tumors. Our study will provide critical insights into normal B cell differentiation as well as lymphomagenesis mediated by IRF4 dysregulation and may uncover novel therapeutic targets.

NIH Research Projects · FY 2025 · 2025-07

Title: DNA aptamers as tools for toxin delivery to glioblastoma Despite recent advances in understanding the pathophysiology of glioblastoma (GBM), translation into improved clinical outcomes is absent. With a 5-year survival rate of less than 5%, nearly every patient with GBM will experience recurrence and succumb to the disease. Therapeutic efficacy is hindered by obstacles including the highly selective blood brain barrier (BBB), and inherent tumor characteristics such as high intratumor heterogeneity, which enables therapeutic resistance and recurrence. Aptamers are short, synthetic, single strands of DNA or RNA. Folding into three-dimensional shapes inherent to oligonucleotides, aptamers act as nucleic acid antibodies, binding ligands with equilibrium dissociation constants in the nanomolar region, comparable to antibodies. Aptamers are also ~6-fold smaller than antibodies and some have the potential to cross the BBB. Aptamers are identified through Systematic Evolution of Ligands by Exponential Enrichment (SELEX) and preliminary experiments suggest that for aptamers to be developed as successful aptamer drug conjugates (ApDCs), training of drug conjugates to properly localize and release the free drug must be included in the SELEX process itself. Thus, this study seeks to optimize approaches to identify DNA aptamers that can be used as GBM-specific ApDCs. Overall question: Can we develop new drug conjugates that deliver therapy to GBM tumors otherwise resistant to treatment due to the blood-brain-barrier and intratumoral heterogeneity? Central Hypothesis: DNA aptamer-drug conjugates can be selected from a vast random library for their ability to home to human GBM patient-derived xenograft (PDX) tumors in mice. Aim 1 will identify GBM-specific MMAE- conjugated DNA aptamers in PDX models. Aim 2 will select ApDCs for intracellular MMAE toxin delivery. Long term goal: Develop a diverse library of therapeutically effective anti-GBM ApDCs from which personalized ApDCs cocktails could be selected to treat GBM patients. Completion of this study will impact the fields of both aptamer technology development and neuro-oncology, while potentially identifying novel therapeutic modalities. Additionally, the research aims of this proposal, coupled with my education plan, provide a unique training opportunity for me to translate my preliminary basic science data into a potential GBM therapy. Not only will the proposed fellowship train me to become an expert in the field of ApDCs, but the project and training environment will also take me one step closer to becoming an effective physician scientist.

NIH Research Projects · FY 2025 · 2025-07

ABSTRACT Arthrofibrosis is a debilitating condition that limits joint motion and causes substantial pain. It is a common complication (~5%) following routine total knee arthroplasty (TKA) and its symptoms are often resistant to pharmacological treatments, physical therapy, and revision surgeries. While aberrant extracellular matrix generation is the primary manifestation, the cellular and molecular effectors that trigger and sustain arthrofibrosis have yet to be elucidated. As a result, there are no therapeutic interventions available to prevent or treat arthrofibrosis following TKA. It is thus critical to establish non-surgical preventive and therapeutic modalities for arthrofibrosis. The scientific premise of our study is that routine surgical trauma caused by index TKA alters the local cellular environment, leading to loss of adipocytes and adipokines that inhibit myofibroblastogenesis and the formation of excessive fibrotic tissue in the joint. This application will address the central hypothesis that activation of the adiponectin cascade suppresses myofibroblast-induced scar tissue formation in the knee. Our working model is that arthrofibrosis development is promoted by the local absence of the adipose-derived hormone adiponectin. Preliminary data from our group reveal that adiponectin expression is suppressed in arthrofibrotic knees and the synthetic adiponectin mimetic AdipoRon exhibits anti-arthrofibrotic properties in vitro and in vivo. Our studies will (i) assess whether local sustained release of AdipoRon can prevent the onset of arthrofibrosis in a pre-clinical rabbit model (Aim 1) and (ii) define the impact of adipogenic factors, including Adiponectin and its mimetic AdipoRon, on myofibrogenesis and decipher the mechanisms by which the adiponectin-axis counteracts myofibroblast differentiation (Aim 2). These studies are impactful as development of novel and non-surgical therapeutic measures that prevent and/or treat arthrofibrosis will reduce complex, unpredictable, painful, and costly revision TKAs. Our studies provide conceptual innovation as a pre-clinical model of arthrofibrosis will be used to test if local AdipoRon delivery in the knee can prevent disease onset. In addition, we will define the role of the adipogenic milieu and of adiponectin signaling activation on myofibroblast differentiation using primary human cell culture models. Overall, we aim to establish novel therapeutic approaches to combat the onset and progression of arthrofibrosis.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY/ABSTRACT Patients with neurodegenerative disease and dementia frequently report difficulties writing in their daily lives as an early symptom, and clinical observations suggest that this may be a valuable tool to support differential diagnosis. Nevertheless, the reported challenges often go overlooked by clinicians who do not have the domain expertise to recognize their relevance or identify their etiology. Empirically, written language production is vastly understudied in these populations, particularly when compared to auditory language and speech production, which also limits the clinical utility at present. We propose that using digital tools to collect and characterize handwriting samples could increase access for timely and accurate dementia diagnoses, particularly for those patients who do not have access to the costly neuroimaging and multidisciplinary expertise that are currently required. Accurate diagnoses are paramount for tailoring prognosis and access to increasingly available disease modifying drugs. Before digital handwriting analysis can inform timely and accurate dementia diagnoses or prognosis, it is crucial to characterize the handwriting patterns specific to each of the atypical Alzheimer’s disease (AD) and frontotemporal lobar degeneration (FTLD) variants. Prior dementia research has focused on linguistic knowledge by examining spelling errors or on singular aspects of fine motor skill in specific groups (e.g., micrographia in Parkinson’s disease). Exploiting informative elements of handwriting samples requires more comprehensive analysis of the writing process and the written product and direct comparison of handwriting across dementia phenotypes. Writing process variables (e.g., stroke speed) are those extracted during the physical act of writing and must be collected using a digitizing tablet or similar tool. Written product variables (e.g., letter size and spacing) can be extracted from any written sample. This proposal seeks support for the technology, personnel, and statistical support to 1) retrospectively analyze longitudinal changes in written product variables, derived from descriptions of the Western Aphasia Battery picnic scene from a cohort of 90 patients with primary progressive apraxia of speech or agrammatic primary progressive aphasia (PPAOS/agPPA) and 40 healthy controls, with at least two annual visits and 2) characterize the written product and writing process in the same task performed on a digitizing tablet in a prospective cohort of 80 patients with atypical AD and FTLD phenotypes (20 each: posterior cortical atrophy, logopenic primary progressive aphasia, PPAOS/agPPA, and semantic dementia) and 60 healthy controls. Achieving these aims would be a foundational step toward developing a digital marker of neurological impairment across AD and FTLD phenotypes, paving the way for earlier, accurate diagnoses in a highly cost-effective and accessible way.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY TAR DNA-binding protein-43 (TDP-43) aggregation is seen in neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). These diseases present a significant burden to patients and caregivers, with little promising intervention to date. Interactions between oligodendrocyte precursor cells (OPCs) and microglia are increasingly recognized to play a role in disease states. OPCs are able to differentiate into oligodendrocytes, the primary myelin-forming cells of the central nervous system (CNS). However, they are also involved in modulating activity of immune cells such as microglia, the principal immune cells of the CNS. In models of neuroinflammation and neurodegeneration, OPCs play a neuroprotective role in attenuating detrimental microglial proliferation. Understanding the mechanistic details of this OPC-mediated neuroprotection has therapeutic implications. The objective of the proposed research is to determine the mechanism underlying OPC-microglia interactions in TDP-43-related neurodegeneration and outline the functional significance in terms of disease outcome. Our preliminary data in the rNLS8 mouse model of inducible TDP-43 expression suggests that OPC-microglia interactions significantly increase during disease. Performing predictive ligand-receptor analysis of transcriptomic data from human ALS patients highlights microglial secreted phosphoprotein 1 (SPP1) signaling to OPC integrin receptors as a likely mechanism. We hypothesize that increased OPC-microglia interactions occur through enhanced SPP1 signaling during TDP-43-related neurodegeneration and serve a protective role by reducing microglial numbers and neuroinflammation. To accomplish our objectives, we will use transgenic and viral mouse models of TDP-43-related neurodegeneration. We will first address whether SPP1 signaling to OPC integrin receptors mediates this interaction through immunohistochemical (IHC) staining and use of an SPP1 genetic knockout mouse model. To investigate real-time dynamics and ultrastructure of this interaction, we will perform in vivo two-photon imaging and serial block-face scanning electron microscopy. We next address the effect of OPC-microglia interactions on disease progression through a pharmacogenetic OPC ablation mouse model. Following ablation, we will measure readouts of neuroinflammation, neuropathology, and motor behavior function. Finally, we will confirm the translational relevance of our findings by performing IHC staining to assess OPC-microglia interactions in human ALS and FTD tissue. Successfully completing these aims will unveil a critical regulatory axis of neuroinflammation in TDP-43-related neurodegeneration, providing a foundation for targeted therapeutic interventions.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY/ABSTRACT Disorders of the inner ear manifest as an array of audiovestibular diseases and constitute a significant public health burden, affecting millions of individuals and costing billions of dollars each year. As the population ages worldwide, the disease prevalence is intensifying, necessitating immediate action to address this growing public health issue. CT imaging of the inner ear is a central component of the diagnostic evaluation of patients presenting with vestibular symptoms, hearing loss, autophony, and chronic otitis media. As a part of this evaluation, accurate detection and characterization of potential labyrinthine dehiscence is imperative to definitively rule in or rule out disease, inform patient counseling, guide shared clinical decision-making, quantify the risk and potential outcome associated with surgical intervention, and further understand the natural history of disease. Notably, current in vivo imaging techniques are unable to consistently decipher thin versus truly dehiscent labyrinthine bone. Further, current CT imaging is not able to fully characterize dehiscence morphology (shape, size, and surface area). As such, high-quality, high-resolution, artifact-free in vivo imaging is critically needed for the reliable diagnosis and management of inner ear disorders such as superior semicircular canal dehiscence syndrome and cholesteatomatous labyrinthine fistula. To address this unmet clinical need, we propose to develop high-fidelity clinical imaging techniques for the inner ear using photon counting detector computed tomography (PCD-CT) and advanced artificial intelligence (AI) algorithms. Our team has been at the forefront of PCD-CT development and its clinical translation, and has an established track record of strong multidisciplinary collaborative innovation in this space. Building upon our successful prior work, we will harness the power of AI – in synergy with PCD-CT – to reduce image noise, improve spatial resolution, and reduce radiation dose for CT imaging of the inner ear. We will develop and validate these techniques with cadavers, and in vivo patient CT exams. The innovation of our proposal lies in the synergy of PCD-CT and advanced AI algorithms for spatial resolution improvement, noise and dose reduction, and a cadaver library of normal and labyrinthine thinning and dehiscence specimens that will enable iterative protocol optimization of ultra-high-spatial-resolution inner ear imaging. Without the proposed techniques, PCD-CT, while impressive, remains limited by noise, image artifacts, and radiation dose. Further, under controlled conditions using cadaveric specimens, definitive evidence of the impact of the developed techniques will be obtained. Successful completion of this proposal will have a significant impact on patients with inner ear diseases, allowing high-fidelity imaging with improved diagnostic accuracy and individualized patient management. In addition, the technical innovations for noise reduction, super-resolution, and dose reduction developed in the proposal will benefit inner ear imaging tasks beyond dehiscence, including direct visualization of the interscalar cochlear partition, and can be readily applied to the middle ear.

NIH Research Projects · FY 2026 · 2025-07

ABSTRACT Obesity prevalence continues to increase in our country and worldwide. The incretins, GIP and GLP-1, facilitate disposal of ingested nutrients and lead to glucose-dependent insulin secretion or inhibition of glucagon secretion. Medications targeting these receptors are effective in inducing weight loss; however, they are frequently associated with gastrointestinal (GI) adverse events (AEs), particularly nausea, and possibly development of gastroparesis. We previously demonstrated that the daily-administered GLP-1 receptor agonist (GLP-1RA), liraglutide (3mg SQ), slowed gastric emptying of solids (GES) and induced mean weight loss of 5.6kg at 16 weeks. Among patients treated with liraglutide, 60% had documented GES T1/2 >174 minutes (“gastroparesis range”) at 5 weeks and 30% at 16 weeks, suggesting that tachyphylaxis occurs with continued treatment with the daily-administered GLP-1RA liraglutide. GLP-1RA treatment has been associated with risk of lung aspiration during endoscopy, suggesting clinically relevant slowing of GES. The overall hypothesis is that pharmacological incretin-based agents administered weekly, semaglutide (GLP-1RA) and tirazepatide (dual GLP-1 and GIP receptor agonist), significantly retard GES, but the effect on GES is heterogeneous, associated with tachyphylaxis, and may persist after cessation of treatment with these agents. Effects of the weekly administered semaglutide on GES beyond 4 weeks’ treatment is unknown. Tirzepatide administered for 4 weeks delayed GE of liquids; effects on GES are unknown. GES is more relevant for risk of pulmonary aspiration with these medications. The effects of semaglutide and tirzepatide on gastric functions (accommodation and emptying of solids) and on GES at 4 weeks after stopping these medications are unknown. We propose one specific hypothesis and aim in obese/overweight adults eligible for these medications. Hypothesis: Semaglutide or tirzepatide result in slowing of GES and increased satiation with considerable inter-individual heterogeneity and evidence of tachyphylaxis. Specific Aim: To compare effects of weekly SQ semaglutide 2.4mg SQ, SQ tirzepatide 10mg, and placebo administered for 24 weeks on GES measured repeatedly at baseline, 16 weeks, 24 weeks, 28 weeks, 4 weeks after stopping the medication, and accommodation and satiation at 24 weeks compared to baseline. Significance: Our study will document heterogeneity in the effects of semaglutide or tirzepatide on gastric motor functions and satiation, the potential for tachyphylaxis in the effect on GES or the persistence in slowing of GES after stopping treatment. This pilot trial will also provide initial data on the coefficient of variation of the effects on GES and satiation in response to the 2 drugs in order to develop the power statement for future clinical trial comparison of semaglutide and tirzepatide.

NIH Research Projects · FY 2025 · 2025-06

Project Summary/Abstract Genetic variations significantly contribute to orofacial cleft. Identifying predisposition genes of orofacial cleft holds promise for early diagnosis, preventive measures, and targeted therapies. However, pinpointing genes harboring rare causal variants remains a challenge due to limited statistical power associated with typical sample sizes of hundreds or thousands of cases and controls. We propose utilizing genetic burden analyses with large control sample size and gene expression data sets to discover orofacial predisposition genes. This approach leverages multiple genomics datasets: ~1,400 orofacial cleft samples from Gabriella Miller Kids First Pediatric Research Program and ~730,000 samples included in the public biobank-level summary counts from the recently released Genome Aggregation Database V4, ~240 orofacial cleft samples and ~220K controls from All of Us Genomics data sets. Furthermore, transcriptomics data sets will be analyzed to enrich potential causal genes, including the analysis of both bulk and single cell RNA-seq data sets from craniofacial tissues and reference GTEx tissues. We will perform discovery analysis, followed by further validation and combined analysis. This comprehensive approach aims to identify novel predisposition genes and variants associated with orofacial cleft, ultimately facilitating early diagnosis, risk stratification, improved understanding of disease etiology, and developing targeted treatments.

NIH Research Projects · FY 2026 · 2025-05

PROJECT SUMMARY / ABSTRACT The social and economic implications of dementia is greatest in women because of their longer life expectancy and resulting elevated risk for dementia compared to men, and this risk of dementia in women may be, in part, modulated by ovarian hormones. Although premenopausal bilateral oophorectomy (PBO) is associated with a reduced risk of ovarian and breast cancer, it is also associated with an increased risk of dementia and other neurological diseases later in life. It is estimated that one in ten U.S. women have undergone abrupt endocrine disruption by having their ovaries removed before reaching natural menopause and this rate is even higher in African American/Black (AA) women. For the large number of women who underwent prophylactic PBO over the past decade, it is critical that the pathologic mechanisms by which PBO influences the risk of dementia should be determined in a rigorous manner for preventive approaches. The most common pathologies that contribute to cognitive impairment and dementia are Alzheimer's disease (AD) and cerebrovascular disease. Our goal is to understand the effects of abrupt disruption of ovarian hormones before the onset of menopause on imaging biomarkers of AD and cerebrovascular disease pathophysiology later in life. We will enroll 150 women who underwent PBO and 150 women who did not undergo PBO drawn from a well-characterized and established population-based cohort. Because our current cohort is predominantly White in Mayo Clinic Rochester, we plan to enrich this cohort with a new AA cohort from Mayo Clinic Florida to reach a sample of 400 women (200 with and 200 without PBO). We hypothesize that imaging biomarkers of AD and cerebrovascular disease, and cognitive function will be abnormal both cross-sectionally and longitudinally in women who underwent PBO compared to referent women who did not undergo PBO, and that this difference will be modified by race (AA versus White), age at PBO, and APOE ε4. Findings from the proposed project have implications both for research and for clinical practice. Findings will contribute to clarifying the mechanisms underlying the development of dementia in women who underwent PBO. In addition, they hold the potential to alter the clinical practice paradigms related to PBO for non-malignant indications and outside of the setting of high genetic risk of ovarian cancer. For the one in ten women currently facing the decision to undergo PBO, determining the late life effects of PBO on the brain is critical for maintaining cognitive health in the long-term.

NIH Research Projects · FY 2025 · 2025-04

ABSTRACT This application requests partial funding for the 2025 Symposium of the Midwest Aging Consortium (MAC). The conference will be held April 23-25, 2025, at the Hilton Rochester Mayo Clinic Area Hotel in Rochester, MN. The expected attendance is 150-200, including basic scientists, physician-scientists, clinicians, and other health professionals. Speakers and attendees will integrate junior and senior investigators and trainees specializing in various scientific disciplines (cell biology, cancer, genetics, medicine, and geriatrics). This conference represents the 6th annual MAC meeting with a unique blend of cutting-edge science in the field of aging. The specific topics to be covered at this conference include: (1) Cellular Senescence and Aging; (2) Age-Related Disease; (3) Metabolism and Aging; (4) Immune Aging; (5) Clinical Research and Aging; (6) Stress Response and Aging. We believe that this meeting is unique, timely, and significant because it will bring together recognized experts from more than ten institutions in the Midwest, including the Mayo Clinic, University of Wisconsin, Minnesota, Michigan, Iowa, Indiana, Northwestern, North Dakota, Ohio State, and Illinois-Chicago, which are all leading institutions in aging, cell biology, genetics, and geriatrics. This 6th MAC symposium is designed to disseminate new information and promote the interaction between established and junior investigators. This meeting will provide a framework for novel approaches and future age-related disease research directions.