Mayo Clinic Rochester

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY Nonalcoholic fatty liver disease (NAFLD) affects 25-30% of US adults and has a major impact on healthcare burden and population health due to higher morbidity and mortality than the general population. Even though NAFLD can progress to cirrhosis, decompensation, and liver cancer, these outcomes affect a small proportion of patients, but there are no accurate methods that are accessible in primary care to identify these patients early. The heterogeneity of clinical phenotypes, lack of universal screening, and risk-stratification approaches lead to delayed diagnosis, phenotype-specific prophylactic or therapeutic intervention, and poor patient outcomes. The long-term goal is to develop easily accessible methods for NAFLD screening and prediction of disease trajectory. The objective of this application is to leverage large electronic health record (EHR) datasets and analytics to enhance the early identification of NAFLD in general healthcare settings. The central hypothesis is that targeted screening with machine-learning (ML)/artificial intelligence (AI) models applied to longitudinal healthcare data (diagnoses, laboratory values, medications, anthropometrics, demographics) can identify predictors of NAFLD risk and, subsequently, a progressive phenotype toward liver events. The rationale that underlies the proposed research is that EHR-based clinical algorithms which identify NAFLD and phenotype the disease trajectory will guide clinicians in selecting patients who need liver-related diagnostic evaluation and enable timely intervention to prevent hard outcomes. Guided by strong preliminary data, the hypothesis will be tested by pursuing two specific aims. In Aim 1, a predictive model for NAFLD will be developed using retrospective data from a population-based EHR-linkage system with validated diagnosed NAFLD and non-NAFLD controls by chart review. Among those with NAFLD, an EHR-based model to predict a clinical phenotype at risk for future liver-related events or death will be developed, using death from other causes as competing risk. In Aim 2, the two models will be validated and calibrated to screen for NAFLD in the population and to predict a liver phenotype at early stage (steatohepatitis or stage≥2 fibrosis). In this aim, the cohort will be prospectively enrolled from the community to undergo NAFLD screening by magnetic resonance imaging and assessment of liver disease severity by biopsy. Lastly, an AI-based NAFLD care model using the predictive models introduced in the EHR system will be implemented in the primary care practice to measure impact on disease identification and management. The approach is innovative because it expands the analytical toolbox beyond conventional methods and challenges the current clinical paradigm which targets only late-stage liver disease identification. These multimodal EHR-based algorithms of NAFLD screening and clinical phenotyping will serve as scalable and unbiased clinical decision-making tools that enable personalized intervention, thereby fulfilling a critical unmet need in the healthcare burden of NAFLD.

NIH Research Projects · FY 2025 · 2024-02

The most prevalent primary brain tumor, glioblastoma, ranks among the most lethal of human cancers. The brain tumor microenvironment (TME) is thought to play a critical role during tumor development and treatment resistance. Unlike many other solid tumors, the glioblastoma TME is dominated by macrophages and microglia— collectively known as tumor-associated macrophages (TAMs). TAMs are plastic in nature and can polarize toward pro-inflammatory or immunosuppressive states. Many lines of evidence suggest that immunosuppressive TAMs dominate the brain tumor microenvironment, which fosters tumor development, contributes to tumor aggressiveness, and impedes the therapeutic effect of various treatment regimens. Through the development of new therapeutic strategies, TAMs can potentially be shifted towards a proinflammatory state to enhance anti- tumor immunity. The promise of TAM-targeted therapy has not yet been realized, due in part to a limited understanding of the molecular mechanisms underlying TAM behavior and function. My postdoctoral work has elucidated novel mechanisms that govern the polarization of TAMs in the glioblastoma TME. Notably, my preliminary data suggests that targeting the CARD9/BCL10/MALT1 (CBM) signaling complex represents a promising therapeutic approach to shifting the glioblastoma TAM phenotype to favor anti-tumor immunity. The overall objectives of this application are to determine the molecular mechanisms that regulate TAM immunoreactivity in glioblastoma and to utilize this information to inform the development of new and effective therapeutic interventions to improve treatment outcomes. My central hypothesis is that CBM activation within TAMs is required for glioblastoma-induced TAM polarization toward an immunosuppressive phenotype and this CBM-dependent TAM polarization facilitates tumor growth, progression, and resistance to therapy. I first propose to elucidate the molecular mechanisms by which glioblastoma cells communicate with TAMs to drive CBM activation (Aim 1). Second, I will evaluate how CBM activity within TAMs influences TAM function (Aim 2). Finally, during the R00 phase of this proposal I will investigate how inhibiting the CARD9-CBM complex in TAMs in vivo affects glioblastoma tumor progression and responsiveness to standard therapies (Aim 3). Collectively, these studies will advance our understanding of the mechanisms governing the brain tumor immune microenvironment of glioblastoma and inform the development of new approaches to manipulating this immune microenvironment to improve treatment outcomes for glioblastoma. During the mentored K99 phase of this award, I will greatly benefit from the expert mentoring world-class research resources available at the University of Pittsburgh and the UPMC Hillman Cancer Center. Completing the proposed project will allow me to build a strong scientific foundation and then lead an innovative research program as an independent investigator. Overall, the K99/R00 award will be an indispensable support for my timely transition to a successful career as a multifaceted, cross-disciplinary investigator in neuro-oncology.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY/ABSTRACT Uveal melanoma (UM) is the most common primary intraocular tumor of adulthood, and radiation-based therapies are the mainstay treatment for most small, medium, and some large UM. Despite high rates of local tumor control, over two-thirds of patients are left with functional blindness in the treated eye due to radiation- induced side effects. Radiation optic neuropathy (RON) is particularly devastating due to its strong association with profound visual deficits and lack of effective treatment options. We recently identified a potential novel opportunity for intervention based on a retrospective study identifying risk factors for RON. In our clinical practice, we observed that some patients with RON manifested with concurrent neuroretinal rim thinning (NRT). Based on this observation, we analyzed a number of potential risk factors for RON in patients treated with plaque radiotherapy for UM and found that higher baseline intraocular pressure (IOP) was associated an increased risk of RON. These data supported a hypothesis previously proposed in glaucomatous optic neuropathies: Specifically, there is a pressure sensitive component to optic nerve injury, which is associated with connective tissue stress and strain at the lamina cribrosa, that results in a cupping or NRT. Few animal models accurately replicate this aspect of human anatomy due to a lack of a well-developed lamina cribrosa. However, the tree shrew has a similar distinct, load-bearing lamina cribrosa, and is rapidly becoming an animal model for studies in conditions including glaucoma and other optic neuropathies. Given our clinical observation and growing use of the tree shrew as a model for optic nerve pathology, our Central Hypothesis is that a novel model of RON can be developed using the tree shrew based on its load-bearing lamina cribrosa, which will replicate characteristic features of human pathology and allow testing of novel therapeutics, specifically those related to IOP reduction. Establishing a representative animal model will have potential to accelerate new treatment approaches for patients with RON. We will evaluate our central hypothesis via two specific aims. Aim 1 will validate the use of the tree shrew as a model for RON by performing a rigorous assessment of the structural and functional phenotype of the tree shrew model of RON. Aim 2 will examine the role of IOP regulation in the severity of RON. Based on strong clinical data with which to form this hypothesis, these aims will support our long-term goal of developing novel treatment approaches including IOP reduction to reduce the risk RON and prevent blindness in patients requiring exposure to radiation.

NIH Research Projects · FY 2026 · 2024-02

Project Summary The developing alveolus is lined by two epithelial cell types: thin alveolar type 1 (AT1) cells that provide the gas- exchange surface, and cuboidal AT2 cells that secrete pulmonary surfactant. During embryonic development both AT1 and AT2 cells derive from a distal progenitor pool. Alveolar epithelial differentiation is influenced by multiple molecular signals –– especially FGF–– and recent work suggests that another key driver in late gestation is mechanical stretch. Questions remain as to how these signals are regulated to precisely time alveolar differentiation (as both are present at and critical for earlier stages of lung development), and to what extent are they reactivated in adulthood during regeneration. Recent work in embryonic stem cells (ESCs) points to a specific cell-intrinsic mechanical property (known to resist cell stretch) that blocks differentiation – cellular membrane tension (CMT). In ESCs, high CMT represses differentiation by blocking the endocytosis of a FGF receptor, which modulates its downstream signaling. While upstream FGF signaling was always active in ESCs, it was the shunting of its downstream signaling that was CMT-dependent and that was critical in timing differentiation. Could CMT also be involved in the embryonic lung to block alveolar differentiation? Does CMT re-emerge in the adult lung during alveolar epithelial regeneration? If so, how does it affect activation of facultative AT2 progenitors? To address these questions, we propose to investigate CMT in both the developing and adult mouse lung. We hypothesize that alveolar epithelial differentiation is blocked by high CMT, which regulates downstream signaling via inhibiting endocytosis and resisting stretch. During alveolar regeneration, we hypothesize that facultative AT2 progenitors are activated by increased CMT that subsequently reduces to direct AT1 and AT2 differentiation. To answer these questions, we will use a combination of culture and transgenic mouse experiments wherein measurements will be taken primarily by confocal microscopy. Our preliminary data indicate that CMT reduces immediately prior to (and is required for) differentiation during development. Concomitantly, we observe an increase of endocytosis prior to differentiation and that its inhibition also blocks differentiation in culture. Finally, we developed two approaches to measure mechanical properties of individual cells within living lung tissue. In doing so, we observed higher CMT in progenitors versus adult AT2 cells, as well as increased stiffness of AT2 cells relative to neighboring cells in the adult lung. Our approach is innovative, as neither CMT nor endocytic regulation of FGF has been studied before in the context of lung development and regeneration. Moreover, we have established two novel approaches to measure mechanics in living lung tissue with cellular resolution. If successful, we will have established a novel mechanism of cell-intrinsic mechanoregulation for the lung field which will be informative in understanding pulmonary diseases with aberrant epithelial differentiation. Further, our findings will guide development of novel diagnostics or therapeutics, either for direct use in patients or in culture to control stem cell differentiation.

NIH Research Projects · FY 2025 · 2024-01

PROJECT SUMMARY/ABSTRACT Heparin is administered as an anti-coagulant to approximately 10 million patients annually, placing it among the most frequently utilized clinical interventions in the US. About 3% of patients that receive heparin generate antibodies that cause heparin-induced thrombocytopenia (HIT). Antibodies that cause HIT are especially dangerous due to their ability to bind platelet/PF4 complexes on the surface of platelets and stimulate platelet activation and thrombosis. Unfortunately, patients with HIT have a very high mortality rate (~10-25%), even with treatment. The crucial knowledge gap limiting the optimal diagnosis and treatment of HIT is that the platelet components that recruit PF4 to the platelet surface to elicit the epitopes recognized by HIT antibodies have yet to be discovered. Aim 1 of this proposal will identify the platelet components that recruit PF4 to the platelet surface and quantify differences in their expression based on age, sex, and ethnicity to better risk stratify patients and identify candidates that can be targeted to improve HIT treatment. Identifying the platelet molecules that recruit PF4 to platelets will also define a set of antigenic targets to improve diagnostic enzyme-based immunoassays (EIAs) for HIT. EIAs are used as the frontline diagnostic tool for HIT but have a poor positive predictive value (~50%), necessitating the use of platelet-based assays to confirm the presence of platelet- activating antibodies. Improving the accuracy of HIT EIAs would eliminate the need to perform platelet-based confirmatory assays and significantly shorten the time to diagnosis and therapeutic intervention. Because only approximately 50% of HIT patients develop thromboses, we hypothesize an inhibitory platelet mechanism limits HIT antibody-mediated thrombosis. As a proof-of-principle, we fractionated platelet granule components and found that glycosaminoglycan-rich proteoglycan fractions specifically inhibit HIT antibody- mediated platelet activation. Aim 2 of this proposal will identify the platelet components that selectively inhibit antibody-mediated platelet activation and will leverage this discovery to develop improved HIT therapeutics. Our preliminary data also identified a subclass of HIT patients who test negative in diagnostic EIAs but positive in platelet activation assays. Because platelet activation assays are typically only performed after a positive EIA, this subclass of patients may be significantly underdiagnosed using current HIT diagnostic workflows. Aim 3 of this proposal will assess the efficacy of current HIT diagnostic strategies by defining the frequency of HIT- suspected patients that test serologically negative in EIAs but positive in platelet activation assays. Aim 3 will also establish the pathogenicity of this HIT antibody subclass in a humanized murine model of HIT. The long-term goals of this study are to better understand HIT pathogenesis, including the age, sex, and ethnicity-driven factors impacting HIT severity, and improve the diagnosis and treatment of HIT by identifying platelet components that 1) recruit PF4 to the platelet surface to elicit the epitopes recognized by HIT antibodies, and 2) participate in an inhibitory feedback mechanism that attenuates HIT antibody-mediated platelet activation.

NIH Research Projects · FY 2026 · 2023-12

Project Summary Prostate cancer is the most common malignancy and the second leading cause of cancer-related mortality in men in the United States. The American Urological Association recommends the use of multiparametric magnetic resonance imaging (mpMRI) for diagnosis and disease management in all men at high risk for prostate cancer. mpMRI is also beneficial in men: 1) with an increasing prostate-specific antigen (PSA) following an initial negative prostate biopsy; 2) during longitudinal follow-up in active surveillance for biopsy- proven low-risk prostate cancer; and 3) for guidance of locally targeted therapies including cryotherapy, high intensity focused ultrasound (HIFU), MR-guided transurethral ultrasound ablation (TULSA), and emerging radioligand targeted treatments for intermediate to high-risk cancers. These recommendations are only realizable through accurate localization and characterization of individual prostate lesions on MRI, which is significantly challenging in men with pelvic metal implants. This is important since the incidence of prostate cancer increases substantially over 50 years of age and in parallel, the prevalence of hip replacement surgery also increases in that age group (estimated to reach ~4 million by 2030 in the US). Most of the metal implants in current clinical use are MR compatible, however, they distort the local magnetic field (B0) causing significant signal loss and image distortion. This is particularly problematic with echo-planar based diffusion-weighted imaging (EP-DWI), rendering these images non-diagnostic. This is a major limitation since DWI, according to the prostate imaging reporting and data system (PI-RADS) classification, is the pivotal sequence for lesion detection and characterization in the peripheral zone, where 70-75% of prostate cancers arise. To address this unmet clinical need, we propose to establish a 3D turbo spin echo (TSE) based DWI that is robust and can be widely implemented across all MR scanners. This is built upon our recently introduced novel acquisition and reconstruction strategy utilizing variable density Cartesian acquisition with spiral profile reordering (VD- CASPR). The volumetric acquisition of 3D VD-CASPR-DWI provides higher signal to noise ratio and the TSE readout improves the robustness to B0 inhomogeneities. This provides superior image quality and enables the visualization of the entire prostate gland without image distortions facilitating accurate detection and characterization of prostate lesions in men with pelvic metal implants. We will optimize the novel 3D VD- CASPR-DWI in phantoms and in-vivo prostate imaging of healthy volunteers. We will then validate this technique in 20 patients without and 20 patients with pelvic metal implants, evaluated for known or suspected prostate cancer. The successful outcome of this project will be an optimized mpMRI with our novel 3D VD- CASPR-DWI that provides pertinent clinical information for better management of prostate cancer patients and subsequently improve their quality of life and overall survival.

NIH Research Projects · FY 2025 · 2023-12

PROJECT SUMMARY/ABSTRACT MLL-rearranged (MLL-r) leukemias account for 5-10% of human acute leukemia and is associated with poor prognosis. The unmet clinical needs and the lack of an effective targeted therapy to the MLL-r leukemias emphasize the need for novel regimens. Recent cancer epigenetics studies discovered a central role for the histone H3 lysine 79 (H3K79) methyltransferase DOT1L in MLL-r leukemogenesis. Important clinical responses have been noted with DOT1L inhibitor treatment as a single agent, however, it is expected that combination treatments will be necessary. Our preliminary studies based on a DOT1L-inhibitor sensitization screen have identified an essential role of the PHF20/KAT8 histone acetyltransferase complex, in supporting the expression of DOT1L-driven oncogenes. The objective of this application is to determine the critical epigenetic mechanisms that collaborate with DOT1L to maintain oncogene expression in MLL-r leukemia. Our central hypothesis is that PHF20 mediates KAT8 recruitment to maintain the locus-specific histone acetylation and transcription of the DOT1L-driven leukemic program. We will investigate the efficacy of DOT1L and PHF20/KAT8 combination therapies (Aim 1), dissect the PHF20/KAT8 chromatin targeting mechanisms (Aim 2), and validate a novel high-density CRISPR protein scan technology for de novo discovery of the functional elements in DOT1L/PHF20/KAT8 (Aim 3). This study is innovative because (1) it introduces a novel concept of simultaneously targeting multiple components of an epigenetic feed-forward loop to efficiently suppress the cancer programs, and (2) it establishes a brand new genetic screen approach for a sub-protein level functional domain discovery. The impact of this research will be of significance because (1) it immediately provides novel therapeutic opportunities against the difficult-to-treat MLL-r leukemias, and (2) it will help identify novel functional elements in epigenetic regulators for future pharmaceutical targeting.

NIH Research Projects · FY 2026 · 2023-12

PROJECT SUMMARY/ABSTRACT Hispanic Americans experience a high burden of type 2 diabetes (T2D) and suboptimal diabetes-related outcomes, including glycemic control, medication adherence, physical activity, dietary quality, and glucose self-monitoring. Interventions that measurably improve these outcomes in culturally relevant ways are needed. Digital storytelling (DST), a narrative-based intervention in which individuals share personal experiences related to T2D, is a promising strategy to support diabetes self-management. Narrative approaches that incorporate community perspectives have been shown to influence health behaviors and engagement more effectively than traditional didactic education. DST interventions may improve diabetes outcomes by increasing knowledge, enhancing motivation, and supporting behavior change; however, their effects on specific, measurable T2D outcomes (e.g., HbA1c, self-management behaviors) have not been well established among Hispanic populations. In addition, it remains unclear how individual characteristics (e.g., age, sex, social support) and baseline clinical and behavioral factors influence response to DST interventions. The overall goal of this F32 proposal is to evaluate the impact of a DST intervention on diabetes-related outcomes and to identify factors that modify intervention effectiveness among Hispanic adults with T2D. Dr. Abby Lohr has training in community-based participatory research focused on improving chronic disease outcomes and seeks to further develop expertise in intervention assessment and tailoring. In collaboration with her mentors, she has developed a comprehensive training plan to: (1) advance mixed methods skills and application of theory; (2) expand expertise in DST intervention development and evaluation; and (3) strengthen skills in translating diabetes research into clinical practice. To achieve these aims, Dr. Lohr will conduct a mixed methods study to (1) quantify changes in key diabetes outcomes (including glycemic control and self-management behaviors) following a DST intervention, and (2) identify sociodemographic and clinical factors associated with variation in response. This work builds upon Stories for Change (5R01DK113999-03), a collaborative research effort between Hispanic community partners and Rochester Healthy Community Partnership. By focusing on measurable outcomes and identifying factors that influence intervention effectiveness, this project directly supports NIH priorities to advance rigorous, reproducible, and solution-oriented research that improves chronic disease outcomes.

NIH Research Projects · FY 2026 · 2023-12

The field of Alzheimer's disease (AD) is entering a new era where increasing numbers of novel treatments targeting the biological underpinnings of AD will be available within the foreseeable future. If we fail to identify cognitive impairment due to AD early, we will miss an important treatment window. There is a critical need for easily accessible, scalable, and sensitive cognitive tools that can aid early detection and monitoring of cognitive impairment and thereby allow earlier intervention to mitigate further decline. High quality, brief cognitive assessment tools that can be deployed remotely or via self-administration in clinic settings represent one key component of the future of AD research and clinical practice. These tools will help enrich clinical trials and aid triage decisions to inform specialty clinic referrals and initiation of treatment in clinics, ideally in conjunction with plasma biomarkers to address both clinical symptoms and underlying biology. Additional validation of novel remote cognitive assessment tools is needed. Mayo Test Development through Rapid Iteration, Validation and Expansion (Mayo Test Drive, MTD) is a cognitive testing platform developed for self- administered digital cognitive assessment. MTD addresses remote assessment needs and is a multi-device (smartphone, tablet, PC), flexible and easily accessible platform, with subtests that provide more in-depth assessment of targeted cognitive domains relative to typical screening tests. The MTD brief cognitive screening battery takes 15-20 minutes and includes 2 subtests: (1) the Stricker Learning Span (SLS), a novel computer adaptive word list memory test (learning and delay trials) and (2) Symbols Test, an open-source measure of visual matching and processing speed/executive function; these are combined into a screening battery composite (MTD-SBC). The overall goal of this R01 is to establish the validity of the MTD-SBC and the SLS for several specific contexts of use. Specific aims are to (1) determine cross-sectional diagnostic accuracy of MTD for clinically defined and PET biomarker-defined groups, (2) demonstrate sensitivity of MTD to amyloid- related cognitive decline over 30 months, (3) determine utility of MTD for detecting clinical progression over 45 months, and (4) determine whether MTD performance is associated with plasma biomarkers to a similar degree as in-person neuropsychological measures. Most participants will be recruited from the Mayo Clinic Study of Aging, with additional recruitment from the Alzheimer's Disease Research Center (Rochester, MN and Jacksonville, FL) and the Memory Impairment and Neurodegenerative Dementia Center - Mayo Clinic Study of Aging at the University of Mississippi Medical Center in Jackson, MS (N=2,300 across cohorts, predominantly remote administration). Including cohorts from Jackson, MS and Jacksonville, FL will broaden sample representation and support exploratory analyses for Aim 4 that will examine the robustness of results. Future research will determine the combined utility of MTD and plasma biomarkers.

NIH Research Projects · FY 2025 · 2023-11

Summary/Abstract The overall goal of this proposal is to better understand the role of the tyrosine kinase receptor EphA2 signaling pathway in alveolar macrophages during Pneumocystis pneumonia (PCP). Pneumocystis jirovecii pneumonia (PJP) remains a significant cause of morbidity and mortality in AIDS1,2. During AIDS and other immunosuppressive states, the absence of CD4 lymphocytic immunity results in exuberant and often fatal PJP3,4. Myeloid cells, particularly alveolar macrophages (AMs) are crucial for anti-PJP innate immunity5-8. The binding of Pneumocystis to alveolar macrophages mediates early host immune response to the fungus9,10. AMs further promote killing and clearance of organisms but are also major sources of proinflammatory mediators contributing to profound pulmonary inflammation and lung injury during PJP11-13. To date, extremely little is known regarding the role of EphA2 receptor and Pneumocystis engagement in mediating subsequent effects on the host immune response and fungal killing during Pneumocystis pneumonia (PCP). We have previously shown the importance of the β-glucan receptor EphA2 in Pneumocystis organism attachment to lung epithelial cells14. Furthermore, we demonstrated that following EphA2 receptor-ligand engagement with Pneumocystis β-glucan carbohydrates, the tyrosine kinase receptor is activated in epithelial cells14. Now, our preliminary studies further indicate the importance of the EphA2 receptors on AMs in inflammatory responses to Pneumocystis. To date, there are no published descriptions of the potential roles of EphA2 on AMs during fungal infection pathogenesis. We now provide exciting new initial in vivo data demonstrating that EphA2-deficient mice have significantly less proinflammatory responses and significantly greater organism burden in both immunocompromised and immunocompetent mouse models of PCP. We therefore hypothesize that EphA2 receptors signal following Pneumocystis binding to alveolar macrophages mediating early host immune recognition and response to the organism. In addition, based on our preliminary in vivo PCP model data, we further hypothesize that EphA2-receptor signaling in AMs is critical for mounting proper lung inflammation and organism control during PCP. Two Specific Aims are proposed. Aim 1: We will characterize the function of EphA2 signaling pathways in AMs following in vitro organism attachment. We will specifically study the resulting cytokine response in AMs, as well as subsequent organism killing by AMs. To address this, we will examine the function of EphA2 in AMs derived from EphA2 (EphA2-/-) receptor knockout and wildtype mice challenged with mouse derived Pneumocystis murina. Specifically, we will study the binding kinetics and inflammatory responses following P. murina interactions with AMs. Initial data from our lab suggests significantly less inflammatory cytokine responses in macrophages from EphA2-/- versus wildtype controls. Furthermore, we will analyze the uptake and killing of P. murina by macrophages, as well as cytokine release by these cells challenged with either P. murina as well as isolated Pneumocystis organism cell wall components. Aim 2: We will further determine the role of EphA2-signaling on modulating lung inflammation, AM subset recruitment, and organism burdens in the PCP immunocompetent and immunosuppressed models in mice. Our preliminary studies demonstrate that CD4-depleted EphA2-/- mice with PCP exhibit significantly increased organism burdens compared to their wildtype counterparts. Accordingly, we will evaluate the time course and magnitudes of organism clearance and inflammatory cytokines in wildtype and EphA2-/- mice. We will directly contrast these parameters in immune competent compared to CD4-depleted mice. Furthermore, we will directly isolate and assess macrophages derived from both groups of infected mice to assess their ability to kill Pneumocystis and generate adequate cytokine responses. We will also employ flow cytometry to further isolate and characterize distinct pulmonary macrophages subset recruitment. This will include whether the AMs are relatively Type 1 or Type 2 polarized and will further assess the recruitment of other macrophage populations include tissue-resident alveolar macrophages (TR-AMs), monocyte- derived alveolar macrophages (Mo-AMs), and interstitial macrophages (IMs). These determinations will be performed in both mouse strains tested in the PCP models proposed. These studies promise valuable insights into the role of EphA2 receptor signaling pathway in macrophages during PCP and should provide initial data that may inform future targeted therapies of this pathway for the treatment of PCP. We anticipate that these studies will define the importance of the of EphA2 receptor signaling pathway in organism killing and regulating the host response in PCP. Modulating these pathways may serve as a novel strategy for therapeutic intervention potentially beneficial to patients with PJP when provided in addition to traditional antibiotic agents.

NIH Research Projects · FY 2026 · 2023-11

Abstract Genomic variants in an individual may be either inherited (i.e., transmitted through the germline) or generated by mutagenesis in post-zygotic cells. Widespread genomic mosaicism in somatic cells of phenotypically normal individuals is now well established. In certain cases, it is known that mutations have causative role in diseases and contribute to neuropsychiatric disorders. But generally, little is known about to what extent natural mosaicism influences an individual’s susceptibility to disease. In our recent study we discovered a phenomenon of hypermutability in adult brains. Hypermutability was not related to diagnosis but increased with age, reaching at least 3% population frequency (95% confidence interval) for brains over 40 years old. The phenomenon of hypermutability could have implications for a number of conditions. Since it correlates with aging, it could be important for age-related neurodegenerative diseases; or, as it involves mutations in cancer-related genes, it may be related to predisposition for brain cancer. Alternatively, hypermutability may reflect imbalances (expansion/contraction) between cell lineages in the developing brain. We have three hypotheses explaining hypermutability. The goal of the proposed study is to firmly establish the origin(s) of hypermutability. To conduct the study, we will first expand the set of hypermutable brains (Aim 1). The ultimate judgement about the origins will be made from the regional distribution and frequency of mutations in the brains (Aim 2), the cell type(s) carrying the mutations (Aim 3), and proving (or disproving) that mutations in the hypermutable brains arose from a clonally expanded cell lineage (Aim 4). Proving the hypothesized origins of hypermutability will be a fundamental contribution to understanding brain development and aging in humans and will lay the foundation for future studies of the underlying mechanisms.

NIH Research Projects · FY 2024 · 2023-09

ABSTRACT/SUMMARY Over 800,000 patients in the United States are suffering from end-stage renal disease, a condition defined clinically as an estimated glomerular filtration rate (eGFR) <15 mL/kg/1.73m2, compared to a normal eGFR >90 mL/kg/1.73m2. This profound loss of renal function makes hemodialysis (HD) treatment, a procedure where the blood is filtered externally to augment renal function, critical for the survival of this patient population. Performing HD efficiently requires access to a section of vasculature with excellent flow, patency, and resistance to repeated punctures. The clinically preferred method of creating such a site is by surgically joining the high-pressure radial or brachial artery with the low-pressure cephalic or basilic vein to create an arteriovenous fistula (AVF). Ideally, once an AVF is created, the outflow vein undergoes a process of arterialization, with a dramatic increase in volumetric flow and an outward thickening that makes a mature AVF an ideal HD access site. Unfortunately, AVFs have a remarkably high failure-to-mature rate, with as many as 40% being unusable 1 year after creation. AVFs can also take months to mature, and their failure probability can be difficult to assess during the maturation process, making it critical to identify biomarkers that are predictive of AVF maturation at the earliest possible timepoints to drive interventional treatments and AVF succession planning. In this project, we propose examining the stiffness of the AVF-adjacent vasculature as one possible class of biomarkers using ultrasound-based pulse wave imaging (PWI). PWI takes advantage of the fact that the speed at which a mechanical excitation propagates through a tube is related to the stiffness of that tube as described by the Moens-Korteweg equation. Such a mechanical excitation is provided continuously by pulsatile flow in blood vessels under in vivo conditions, and can be imaged using ultrasound. However, imaging this wave robustly requires excellent temporal and spatial resolution, as pulse waves in vasculature propagate at speeds on the order of meters per second and typically induce small (<1 mm) deformations, which has made PWI difficult and hampered its clinical adoption. Several new US imaging techniques have recently been proposed that promise to increase spatiotemporal resolution including Time-Aligned Plane Wave Compounding (TA- PWC) and Comb Detection (CD), but these have not yet been applied for US-based PWI. In Specific Aim 1, we propose to apply the TA-PWC and CD techniques to PWI and validate the results in vascular phantoms and both ex vivo and in vivo porcine common carotid arteries to improve the spatiotemporal resolution of ultrasound. In Specific Aim 2, we plan to apply PWI to measure AVF stiffness in porcine models of chronic kidney disease and assess the relationships between past values of stiffness and future surrogates of AVF maturation, the relationship between AVF stiffness and histologically-assessed fibrosis, and the agreement between PWI-assessed AVF stiffness and mechanical testing of excised tissue.

NIH Research Projects · FY 2026 · 2023-09

PROJECT SUMMARY There has been a dramatic increase in the number of persons living with reduced physical function and with aging-related chronic conditions. If we compare chronological age (calendar-based age) with biological age (changes at the cellular, tissue, organ, and system levels), we can classify persons as aging faster (accelerated aging) or slower (successful aging) than their peers. Methods have been developed to measure biological age based on DNA methylation, telomere length, and blood biomarkers. However, such measures may not accurately reflect organ- and tissue-level changes from aging. A multi-organ/tissue approach is needed to identify comprehensive age-related structural changes before signs, symptoms, or clinical diagnoses occur. Abdominal computed tomography (CT) has widespread use in the general population (35% of adults ages 20-89 years in an 11-year period). Quantitative measures of the organs and tissues on abdominal CT may predict organ-specific diseases, or in combination, may be used to calculate biological age and predict the more global outcomes of hospitalization and mortality. Therefore, our central hypothesis is that deep learning (DL) models applied to abdominal CTs can quantify structural features of the organs and tissues to identify persons with accelerated aging at high-risk for organ-specific disease, hospitalization, and death. The Rochester Epidemiology Project record-linkage system provides access to a general population archive of images for 423,081 abdominal CTs and to comprehensive medical record data among 181,187 adults (ages 20-89 years) between 2010-2020. Our team has already developed and validated DL tools to measure liver, kidney, aorta, fat, muscle, and bone on abdominal CT images. We will leverage these resources to 1) establish percentiles of abdominal CT biomarkers from both healthy and general population samples; 2) determine the risk of organ-specific clinical disease by abdominal CT biomarkers in the general population; and 3) determine the risk of hospitalization and death associated with abdominal CT measures in the general population. If successful, application of DL tools to abdominal CT images will enrich the characterization of age-related health risks without additional testing burden. Subclinical abdominal CT biomarkers may also inform the biology of aging and early disease, improve disease classification, and provide opportunities for early intervention.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY/ABSTRACT Covid-19 has increased the prevalence of substance use disorders (SUD) visits in primary care, which, when coupled with problems of mental healthcare access for vulnerable rural populations, accentuates already existing disparities for these patients. There is an urgent need to improve the timely identification and management of SUD for rural primary care patients. Screening, brief intervention, and referral to treatment (SBIRT) is an evidenced-based strategy to address SUD in the primary care setting, but it is seldom fully used. Building on our experience in depression collaborative care (CC) management and ability to scale telehealth outreach, we built a digitally native integrated behavioral health CC platform using a novel app and provider platform called Senyo Health. We will utilize this SUD CC platform to complement existing CC for depression to deliver SUD care integrated into primary care. We feel this approach will enhance SBIRT care and improve patient-centered outcomes. If our pilot succeeds, we will scale the Senyo Health platform for SUD treatment among rural Midwest Mayo Health System patients. With this study, we will apply two operational strengths of Mayo Clinic to a new domain of care delivery: a demonstrated track record of implementing integrated behavioral health CC to improve the management of depression in the primary care setting and a demonstrated ability to rapidly deploy telemedicine service for all psychiatric and psychological care during the COVID-19 pandemic. Through diverse stakeholder engagement, we will explore specific barriers, facilitators, and optimal implementation processes to adapt and pilot test a digitally native CC platform for SUD. Using a mixed implementation and efficacy study designed, we propose enrolling patients from three clinical sites, including two rural health clinics, to evaluate the intervention's acceptability, feasibility, and potential effectiveness. Building off this initial phase, we will expand our intervention to include four more new clinics continuing the same mixed methods to adapt and improve implementation processes and clinical outcomes. We will then disseminate our acquired knowledge through research publications, web stories, and a manualized process to share with those interested in implementing a virtual CC approach to delivering SUD services in primary care.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY / ABSTRACT We propose to improve risk assessment of pancreatic ductal adenocarcinoma (PDAC), a major lethal cancer for which no early detection strategy exists. We hypothesize that a novel enhanced multifactor risk score (eMRS) will improve risk stratification for moderate to high genetic risk strata for PDAC, by incorporating carrier status of hereditary cancer syndrome pathogenic variants. We will develop a new eMRS using large Mayo Clinic datasets of PDAC cases recruited since 2002 and primary care controls in the Mayo Clinic Biobank with recently completed whole exome sequencing (WES) by Regeneron Genetics Center (RGC) to first assess the limited number of existing published models. We will then construct our eMRS with these data and perform independent validation using the UK Biobank. We will test the value of the model in a dataset of at-risk first- degree relatives (FDRs) of Mayo Clinic PDAC patients. Our new eMRS for PDAC will for the first time comprehensively accommodate inherited cancer syndrome pathogenic variants to stratify risk of PDAC in identically processed sequencing and informatics workflows. The study team includes leaders in gene identification and genetic epidemiology of PDAC, having led the GWAS that identified the risk SNPs that will be included in risk model testing and development. By leveraging the three existing major genomic datasets and one family dataset with accompanying epidemiologic and disease covariates, our specific aims are: (1) To use uniquely coordinated datasets on Mayo Clinic PDAC cases and Mayo Biobank controls to validate published polygenic risk score (PRS) and MRS models. We will assemble and analyze Mayo Clinic’s 4,552 prospectively recruited cases and 53,229 controls to evaluate the heretofore unvalidated models. (2) To build and validate a novel, enhanced eMRS for PDAC that incorporates cancer germline pathogenic variants. (2a) We will use the datasets built for cases and controls in Aim 1 and include additional epidemiologic risk factors and germline pathogenic variant status for hereditary cancer genes, which have not been previously available in public datasets. (2b) We will validate our new eMRS with an independent dataset of genotype and risk factor data extracted from 2,371 PDAC cases and the remaining 497,629 non-PDAC UK Biobank participants whose germline DNA will have also been sequenced by RGC. (3) To determine the extent that PRS, MRS or eMRS can further refine PDAC risk in first degree relatives of PDAC cases. We will compare standardized incidence ratios (SIRs) in a dataset of personal cancer histories of 23,739 relatives assembled from Mayo Clinic PDAC cases’ family history questionnaires to quantify and assess impact. This project will result in several useful genetic risk stratification models to provide more precise risk assessment for PDAC. The results are anticipated to have a translational impact on risk assessment, genetic counseling, and early detection of PDAC through better, potentially targeted, identification of individuals at increased risk who in the future could be offered minimally invasive surveillance or other options for early-stage detection and prevention.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY/ABSTRACT Irritable bowel syndrome is a globally prevalent disorder (~11%) characterized by an alteration in stool form/frequency and abdominal pain one or more days per week. Abdominal pain in IBS, like other forms of visceral pain, is often diffuse and poorly localized, making it difficult to delineate the site of pathology, and as a result there are few therapeutic options. Gut microbial products have been shown to be important luminal signals for abdominal pain, but these studies have largely focused on the colon. We recently found that small intestinal microbial composition is associated with gastrointestinal (GI) symptoms like abdominal pain, but the mechanisms underlying the role of small intestinal microbiota/microbial products in the pathophysiology of abdominal pain remains a critical knowledge gap. To address this gap, we will elucidate the sensory innervation of the proximal small intestine and identify the cellular and molecular pathways by which the small intestine detects and transduces luminal microbial signals that contribute to visceral hypersensitivity. In our extensive preliminary studies, we established a dedicated model to study the physiologic effects of human small intestinal microbiome in germ free (GF) mice, metabolite effects on isolated DRGs and epithelial cells, and a novel ex vivo spinal cord- small intestine preparation to study the transmission of luminal signals to the spinal cord via luminal metabolite signaling through (1) EC cells, that are epithelial cells which signal to neurons via serotonin, a neurotransmitter in the gut that modulates visceral pain, and (2) sensory neurons in dorsal root ganglia (DRG), the first-order afferent neurons of pain pathways. Using these models, we identified distinct bacteria and bacterial metabolites that activate EC cells and thoracic DRG neurons. Based on our preliminary findings, we hypothesize that small intestinal bacterial products contribute to visceral hypersensitivity by activating small intestine sensory afferents directly and through neuro-epithelial connections by activating EC cells. We will address the hypothesis in two Specific Aims using cutting-edge stimulation/acquisition approaches, including combination of Ca2+ imaging in organoids, electrophysiology of EC cells and DRG neurons, ex vivo preparations from novel transgenic mice, and adeno-associated viruses (AAVs) characterizing the sensory input from the small intestine, and optogenetics to study neuro-epithelial signaling. In Specific Aim 1, we will determine mechanisms underlying activation of EC cells and DRG neurons, and in Specific Aim 2, we will determine the sensory transduction pathways involved in responding to distinct microbial products. These studies will be the first to provide a functional and molecular characterization of sensory neurons in the DRG that innervate the small intestine, determine which subpopulations are activated and/or sensitized by microbial products in the lumen, and test whether EC cells are involved in the sensory transduction pathway. Our findings will allow the development of novel microbial therapies for abdominal pain that target distinct microbial pathways in the small intestine.

NIH Research Projects · FY 2025 · 2023-09

Multiple myeloma (MM) is the most common blood cancer in persons with a higher proportion of African Ancestry (AA) compared to those with a higher proportion of European Ancestry (EA). It remains unknown why the incidence and outcomes of MM vary by genetic ancestry. Large-scale studies comparing variation of the MM tumor, its tumor microenvironment (TME) and disease survival among populations with differing genetic ancestry are critically needed. Thus, our long-term goal is to identify factors contribution to variation in incidence and survival outcomes across populations with differing genetic ancestry. The overall objective of this proposal is to characterize the genetic variations of the MM tumor, its TME, and the impact of this variation on disease survival in a large, well-powered study of patients with differing genetic ancestry. We hypothesize that AA patients in comparison to EA patients have favorable MM tumor genetics but a greater immunosenescent TME, which can affect response to therapy and overall survival. The following specific aims will be evaluated: 1) Differentiate the genetic variations of MM tumors between newly diagnosed AA and EA patients; 2) Analyze the MM tumor microenvironments of newly diagnosed AA and EA patients; and 3) Compare the responses to treatment of MM tumors in newly diagnosed AA and EA patients. In specific aim 1, we will analyze newly diagnosed MM patients from two independent cohorts and determine the frequency of risk-defining tumor genetic abnormalities, genome-wide genomic complexity, and mutation signatures in association with genetic ancestry. Differences in disease survival will be compared in relation to these risk-defining genetic abnormalities and the influence of genetic ancestry. In specific aim 2, we will analyze newly diagnosed MM patients from Mayo Clinic to characterize the TME signatures using RNAseq and validate using CyTOF. Differences in TME and disease survival will be compared in relation to these TME signatures and the influence of genetic ancestry. In specific aim 3, we will analyze newly diagnosed MM patients from Mayo Clinic to characterize their tumor responses to therapeutic regimens using an ex vivo drug sensitivity platform in association with genetic ancestry. Genetic and transcriptomic predictors of ex vivo drug response will be assessed, and top targets and novel agents will be evaluated using human myeloma cell lines. This proposal is significant because understanding MM tumor genetics and TME across patient populations with differing genetic ancestry will allow for improved treatment selection and prognostication.

NIH Research Projects · FY 2025 · 2023-09

Metastasis is the primary cause of cancer death, yet therapeutic strategies to inhibit metastatic invasion do not exist. The long-term goal of our research program is to define the molecular mechanisms driving metastatic invasion, with the goal of identifying novel therapeutic targets and strategies to improve cancer survival. While tumor cells are known to undergo metabolic reprogramming to support tumor growth, the metabolic drivers of metastasis are poorly understood. This proposed research will define how stored lipids are used as a fuel source to power metastatic invasion in pancreatic cancer. We have preliminary data that pancreatic tumor cells undergo a shift towards lipid storage, and that this is required for invasion. This occurs through a suppression of the hormone sensitive lipase (HSL) by the oncogene KRAS, leading to lipid accumulation and priming tumor cells for metastasis. These stored lipid droplets are then catabolized during the process of invasion via the action of lipases. This results in increased oxidative metabolism in the most migratory cells, thereby coordinating lipid droplet breakdown and fatty acid oxidation with cell migration. These data lead to the hypothesis for this proposed research that PDAC cells undergo a metabolic shift to favor the accumulation and storage of lipid droplets, which are catabolized during invasive migration to fuel oxidative phosphorylation to power metastasis. Using a combination of cell biology, biochemistry, and in vivo models, we will test this hypothesis by defining the mechanisms of lipase suppression leading to lipid droplet storage (Aim 1), and the coordinated and localized activation of lipolysis to drive tumor cell invasion (Aim 2). Successful completion of this research will provide fundamental advances in defining the metabolic pathways regulating invasive migration, with a focus on lipid droplets, and with the goal of identifying metabolic vulnerabilities in tumor cells that will provide targets for therapy to block metastasis and improve survival.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY/ABSTRACT This application investigates the relationship between mitochondrial RNA, inflammation, senescence, and aging. Cellular senescence is a well-established driver of tissue and organismal aging, a process thought to be partly mediated via the induction of a chronic Senescence-associated secretory phenotype (SASP). Consequently, there is great interest in selectively targeting senescent cells as a strategy to promote healthy aging. Our work has demonstrated that mitochondrial dysfunction is a hallmark of cellular senescence and a driver of the SASP and that by targeting them we may be able to suppress the detrimental SASP known to contribute to aging. Mitochondrial double-stranded RNA (mtdsRNA) when present in the cytosol is known to be especially immunogenic and trigger an inflammatory response. We observed that senescent cells contain increased levels of cytosolic mtdsRNA together with increased expression of RNA sensors. Furthermore, we found that transfection of cells with mtdsRNA triggers an inflammatory response and inhibition of mitochondrial DNA transcription in senescent cells, decreases the SASP. Finally, we observed that mtdsRNA requires the cytosolic DNA sensor cGAS to induce inflammation suggesting an interplay between RNA and DNA sensing pathways in the process. This led us to hypothesize that cytosolic mdsRNA plays a role in the regulation of the SASP and may be a novel target for interventions to improve healthspan during aging. In this project, we will dissect the mechanisms by which mtdsRNA increases in the cytosol of senescent cells as well as investigate if activation of RNA sensing pathways contributes to the SASP. Additionally, we will explore the mechanisms mediating the interplay between mitochondrial DNA and RNA in the development of the SASP. Our ultimate goal is to identify new interventions that target senescent cells to alleviate age-related dysfunction.

NIH Research Projects · FY 2025 · 2023-09

ABSTRACT: Continuous colon motility is critical for the overall health and survival of an organism and results from activity in the enteric nervous system (ENS) and interstitial cells of Cajal (ICC) that are electrically-coupled to smooth muscle. Although these cellular components have been individually studied in detail, how they interact to coordinate motility across the length of colon is not well understood. Two motor patterns are measured experimentally: (1) ‘ripple’ contractions produced by ICC slow waves of depolarization, and (2) colon migrating motor complexes necessary for propulsion of fecal contents that require ENS activity. Existing models of colon motility are focused on distal regions where distension from a fecal pellet activates intrinsic sensory neurons (or IPANs) that excite ENS motor neurons for oral contraction and anal relaxation of smooth muscles; the forward movement of the pellet then distends the adjacent segment, activates another IPAN, and this ‘neuromechanical loop’ ensures propagation and propulsion of fecal contents. However, these models do not explain the regular rhythm of colon motor complexes, which occur every 2-5 min, or how they are first initiated in the proximal colon where fecal pellets have not yet formed. Unlike motor complexes that reach distal regions only when sensory input is applied, spontaneous, rhythmic motor complexes occur in proximal regions regardless of luminal content, stretch, or distension, indicating that the proximal colon has unique pacemaker capabilities that determine the rhythm of motor complexes. The objective for this project is to determine and model the cellular interactions unique to the proximal colon that are responsible for generating rhythmic motor complexes in normal and inflamed conditions. We hypothesize that rhythmic motor complexes are due to cyclical interactions among ICC, IPANs and motor neurons of the ENS, and that dysmotility during inflammation is due to dysregulation of these interactions. To test this and address knowledge gaps, we will use optogenetics, calcium imaging, in situ immunofluorescence, and computational modeling to define the cell-to-cell interactions responsible for spontaneous, rhythmic motor complexes produced in the proximal colon and determine the cellular components that contribute to dysrhythmic motility following inflammation. Aim 1 will determine the mechanical sensitivity of proximal colon IPANs to ICC-generated ripple contractions. Aim 2 will define the ‘ENS neural program’ activated by IPANs that produces motor complexes in proximal colon. Aim 3 will determine the effect of ENS activity on ICC slow waves and ripple contractions. Each Aim will collect data from normal and inflamed colons, and findings will be incorporated into our model to computationally test whether predictions can be made regarding motility behavior based on changes in cellular activity. Thus, these studies will yield a novel computational model that will help identify cellular mechanisms of dysfunction in colon diseases and guide optimization of therapeutic devices that employ pacemaker technology or nerve stimulation to normalize and restore colon function.

NIH Research Projects · FY 2025 · 2023-09

PROJECT DESCRIPTION/ABSTRACT Patients with glioblastoma and other high-grade gliomas have a dismal prognosis, and there is a compelling unmet medical need to develop more effective therapies. In conjunction with maximal surgical resection, radiation therapy is a cornerstone of treatment for these patients. While focal radiation therapy significantly improves tumor control, approximately 80% of tumors progress within the irradiated volume. While radiation dose escalation has not appreciably impacted this pattern of local failure, there is a compelling rationale to develop novel pharmacologic strategies to enhance the efficacy of radiation therapy in high-grade gliomas. The focus of this application is the first-in-man clinical evaluation of a highly potent, brain penetrant ATM inhibitor (WSD0628) in combination with radiation in high-grade gliomas. We have developed significant pre-clinical data demonstrating robust radiosensitizing effects in cell culture and orthotopic brain tumor patient- derived xenografts (PDXs) with WSD0628. Importantly, a long-term survival study demonstrated no evidence of enhanced CNS toxicity when WSD0628 was combined with a high, single dose of radiation. This is in contrast to significantly enhanced radiation toxicities in epithelial tissues (skin, oral and gut mucosa) observed with this drug. In contrast to most other peripheral tumors with intimate adjacency to various epithelial tissues, minimal ‘at-risk’ epithelial tissues receive significant radiation dose with high-grade glioma treatment, and this can be further limited by using restrictive radiation therapy planning constraints. While this provides a theoretical rationale to combine WSD0628 with radiation in either newly diagnosed or recurrent gliomas, this first-in-man study will be limited to recurrent patients with an especially dire prognosis where a higher risk to benefit ratio is clinically appropriate. As a first step towards understanding a biologically effective tumor tissue concentration in humans, we will use multiple orthotopic GBM PDXs to develop a PK→PD→efficacy model to describe total and free-drug WSD0628 plasma and tumor concentrations associated with robust ATM inhibition and radiosensitizing effects. This model then will be used to interpret the systemic PK data collected as part of a WSD0628 dose-escalation and dose-expansion Phase I clinical trial. In addition, six patients requiring surgery will be treated on the same day with WSD0628, radiosurgery, and surgical resection. By carefully integrating the pre-clinical and clinical data, this study will provide a biologically-informed recommended Phase 2 dosing regimen for effective radiosensitization by WSD0628 for treatment of recurrent high-grade glioma.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY/ABSTRACT – NETWORK COORDINATING CENTER The Network Coordinating Center (NCC) will facilitate and be responsible for network-wide coordination and harmonization of scientific and clinical needs of the funded UG3/UH3 centers comprising the Cancer Adoptive Cellular Therapeutics (Can-ACT) Network. The Network Coordinating Center at Mayo Clinic (NCC-MC) will serve as the administrative hub and coordinating center for all network-related trials and projects. This will include multiple aspects of the functioning of this network, including the administrative and scientific coordination of the Can-ACT Network. The administrative coordination will include (a) providing leadership and facilitating interactions across the network, (b) providing administrative infrastructure to support the activities of the Can-ACT Network, and (c) the coordination of the restricted collaborative funds and administrative supplements for the network. Scientific coordination of this network will include several different critical components, including (a) identifying and establishing strategies for facilitating reagent and specimen sharing, (b) identification and harmonization of process development and manufacturing steps for cell product management and utilization for the functioning of the network between centers and the ICN Core, (c) biostatistical support and collaboration for preclinical and clinical research studies and centralized data management and shared data governance, and (d) bioinformatics and data science support and collaboration. The infrastructure, expertise, and experience of the individuals at Mayo Clinic with national coordination of translational and clinical research is extensive, making this group well-positioned to function as an outstanding coordinating center for the Can-ACT Network. The leadership of this proposed coordinating center brings with them complementary and deep experience and background across these key aims, and they have worked extensively with multi-institutional trials, consortia, and networks as well as cooperative groups in research leadership, trial coordination and implementation, data structures, and importantly development of SOPs and guidance for newly formed collaborative research networks across multiple institutions. Beyond the experience and expertise, the NCC-MC also leverages institutional infrastructure and resources to adapt and evolve as the Can-ACT Network develops and grows through the expected and necessary collaborations across all UG3/UH3 centers and project leaders.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY Approximately 10% of neonates require assistance to begin breathing after birth. At-risk neonates, i.e., those requiring advanced resuscitation due to prematurity, brain injury, or other conditions, are at increased risk of mortality when born in a hospital without a neonatal intensive care unit (‘outborn’) compared to at-risk neonates born in a hospital with a NICU (‘inborn’). Furthermore, at-risk outborn neonates are more likely to suffer serious morbidity, e.g., pneumothorax, severe intraventricular hemorrhage (sIVH), and seizures, and receive cardiopulmonary resuscitation in the delivery room. We estimate that there are 10,000-15,000 at-risk outborn neonates each year who experience poorer health outcomes due to their birth location. However, there are limited strategies to optimize the resuscitation of at-risk neonates born in community hospitals that lack higher levels of neonatal care. There is a critical need to improve patient-important outcomes for this population. The overall objectives in this application are to (i) determine the impact of real-time, audio-video telemedicine consults provided by a neonatologist (termed teleneonatology) on the risk of early mortality and morbidity for at-risk outborn neonates, and (ii) evaluate the effect of teleneonatology on delivery room care provided to these neonates. The central hypothesis is that teleneonatology reduces early mortality and morbidity and improves delivery room care for at-risk outborn neonates. The rationale for this project is that teleneonatology brings resuscitation expertise to the bedside of at-risk neonates more effectively than a brief telephone consult, which is the current practice. Through teleneonatology, the neonatologist can visualize the neonate and provide step- by-step guidance to the community hospital care team. The central hypothesis will be tested by pursuing three specific aims: 1) Determine the impact of teleneonatology on the risk of early neonatal mortality (death within 7 days), 2) Identify the effect teleneonatology has on the risk of early morbidity (defined as pneumothorax, sIVH, or seizure during the first 7 days of life), and 3) Evaluate the impact of teleneonatology on delivery room care. The three aims are embedded into a single prospective, multi-center research trial. With sequential roll-out of the intervention using a cluster randomized stepped wedge design, neonatologists from four NICU ‘hub’ sites will provide teleneonatology consults to at-risk outborn neonates born at 27 community hospital ‘spoke’ sites. Completion of the proposed research will contribute the first-ever, high quality evidence on the impact of teleneonatology on neonatal health outcomes. The research is innovative because it uses video telemedicine to connect neonatologists exactly when needed to at-risk outborn neonates in a way that was not previously possible. This project is significant because it may identify an innovative care model that reduces the long- standing outcome disparities experienced by at-risk outborn neonates.

NIH Research Projects · FY 2025 · 2023-09

SUMMARY Visual perception is a confluence of visual input and attentional state. How visual inputs are processed and represented in human visual cortex is being captured with increasing accuracy in the field. However, a critical gap in our understanding is how top-down inputs from higher order areas influence and sculpt sensory processing—especially an understanding grounded in direct causal manipulations. The long-term goal of the proposed research is to develop causal perturbation methods that modulate visual processing in ways that mimic attentional processes. To achieve this goal, we will use electrical stimulation to test, refine, and extend a quantitative model of feedback information flow to the ventral temporal cortex (VTC). Our research leverages a recent clinical shift in epilepsy surgery from brain surface recordings with electrocorticography to stereo-electroencephalographic (sEEG) depth electrodes: sEEG provides unique opportunities to measure VTC and electrically stimulate feedback projections from gyral and sulcal parietal, frontal and anterior temporal regions as well as the white matter pathways that connect these areas to VTC. In Aim 1, we will determine how stimulating these different feedback locations affects neuronal activity in VTC and simulate VTC signals in a model of a cortical column. VTC electrodes will be localized functionally with a scene perception task and structural connections will be identified from preoperative diffusion MRI measurements on Mayo Clinic’s novel compact 3T scanner with high performance gradients and distortion free imaging. In Aim 2, we will determine the optimal stimulation frequencies for feedback pathways to VTC. Electrodes in feedback pathways will be stimulated with different temporal frequencies and measurements in VTC will be used to characterize optimal response frequencies. In Aim 3, we will test predictions of a previously proposed model of feedforward drive and feedback modulation in VTC. We will present visual scenes while delivering electrical stimulation of feedback pathways and measure changes in sEEG neural responses and behavioral reaction times. This combination of electrical stimulation and visual perception allows us to test and refine an extant model of how interactions between VTC and the intraparietal sulcus affect visual processing and contribute to visual perception. Overall, this multidisciplinary proposal will deliver a set of empirical measurements and models that describe the causal influence of feedback pathways on human VTC. These studies will provide the groundwork for future research to design perturbation based strategies that mimic human attentional conditions.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY/ABSTRACT Heterotopic ossification (HO), a diverse pathologic process of formation of extraskeletal bone in muscle and soft tissues, can be classified into hereditary and nonhereditary (acquired) HO types. A rare and devastating form of hereditary HO is fibrodysplasia ossificans progressiva (FOP), caused by gain-of-function mutations in Activin receptor A type I (Acvr1) gene. In the nonhereditary forms, HO frequently occurs in tendons and ligaments and is commonly incited upon soft tissue trauma, orthopedic surgeries, combat-related blasts, and burns. There are currently still no effective drugs to treat HO. Our long-term goal is to decipher the mechanism(s) of HO and develop novel strategies for the clinical treatment of HO. The overall objectives are to (i) elucidate the essential roles of cilia on the Bone morphogenetic protein (BMP) signaling pathway and (ii) determine its function using novel HO models. The central hypothesis is that primary cilia play central roles in transducing normal and pathogenic BMP signaling and regulating the pathophysiological mechanism of both hereditary and nonhereditary HO. The rationale is that both hereditary and nonhereditary HO formation are primarily mediated by the BMP signaling pathway. However, it remains elusive where and how BMP signaling is transduced and regulated in cells. Primary cilia are antenna-like structures protruding from the surface of cells and are critical for proper transduction of many cellular signaling pathways. Dysfunctional primary cilia result in a broad spectrum of human diseases collectively termed ciliopathies. Our preliminary data suggest that primary cilia play central roles in transducing normal and pathogenic BMP signaling and regulate the pathophysiological mechanism(s) of HO formation. The central hypotheses will be tested by pursuing these specific aims: 1) Determine the ciliary components/pathways that govern normal and pathogenic BMP signaling pathways. 2) Determine the regulatory function of cilia in our newly established nonhereditary burn/tenotomy-induced HO mouse models that have abrogated ciliary signaling pathways. 3) Identify targets for inhibition of cilium-related pathways as the basis for treatments in hereditary HO and nonhereditary HO. The research proposed in this application is innovative because our preliminary data suggest that FOP and perhaps other HO conditions represent cilium-mediated disorders, which provides a novel perspective for future therapies. Mechanistically, we will systematically elucidate the function of cilia on BMP signaling specifically as associated with HO mouse models. The proposed research is significant, and scientifically it will build a new paradigm that primary cilium plays a central role in transducing BMP signaling in chondrogenesis and/or osteogenesis that ultimately results in both hereditary and nonhereditary HO. Clinically, targeting the cilia-mediated BMP pathway is an unexplored therapeutic strategy that could be applied not only for FOP but also for other more common forms of HO, such as post-traumatic HO.