University Of Minnesota

NIH Research Projects · FY 2026 · 2026-06

Our primary goal is to acquire new insights into chronic GVHD (cGVHD) pathobiology to create new therapies to limit fibrosis. We’ve shown that T:B cell engagement can initiate cGVHD by causing pathogenic αhost immunoglobulin (Ig) deposition that exacerbates tissue injury, recruits monocytes (monos) and TGFβ-secreting macrophage (Macs), and stimulates fibroblast/myofibroblast/endothelial cell pro-fibrinogenic cytokines. Our central hypothesis is that developing effective αfibrotic therapies requires greater elucidation of tissue cellular mechanisms and dynamic evolution processes that culminate in cGVHD. We will use state-of-the-art techniques in valid mouse models to expose cGVHD vulnerabilities. Tissues will be obtained at an early and late timepoint from cGVHD mice with bronchiolitis obliterans (BO) or scleroderma (Scl) to assess cGVHD progression in lymphoid and cGVHD organs. In an innovative, scientifically and technologically unprecedented, approach we will interrogate cGVHD mechanisms in BO and Scl models by spatially and temporally resolving and integrating proteomics with whole genome transcriptomics at a single cell resolution within histopathological regions of interest. This will result in a spatiotemporal atlas mapping how specific cells drive cGVHD disease progression in both lymphoid and target tissues; an invaluable tool for future studies. Our specific aims will test the hypotheses that: Aim 1. Interrogating T cell:B cell crosstalk at cGVHD tissue sites will lead to novel therapeutics and individualized applications. During the mechanistic discovery phase, we will infuse bifunctional (suppressive and cytolytic) αCD19 scFv chimeric antigen receptor (CAR19) Tregs to preclude B cell support of pathogenic IgG secretion, leveraging our murine cGVHD/BO cell atlas to assess the means by which these cells disrupt cGVHD progression. Aim 2. cGVHD tissue injury recruits monos that evolve into αinflammatory, pro-fibrinogenic Macs and engagement with fibroblasts/myofibroblasts to initiate fibrosis. Coupling mono and Mac reporter and deleter donor mice with spatiotemporal multi-omics, we will define the mechanisms by which monos and Macs enter cGVHD tissues and pro-fibrotic cytokines are produced, leading to new and key therapeutic targets. To halt fibrosis, mannosylated lipid nanoparticles with TGFβ1 siRNA will be given to selectively bind CD206+ Macs linked to murine cGVHD/BO and Scl. Aim 3. Mac communication with fibroblasts/myofibroblasts causes fibrosis that can be halted by fibroblast activation protein (FAP) CAR Tregs. Utilizing our first of its kind cell atlas of disease progression, we will elucidate the nature of crosstalk between profibrogenic TGFβ-producing Macs, fibroblasts, myofibroblasts and endothelial cells culminating in tissue fibrosis. We show FAP upregulation in cGVHD lung (BO), skin (Scl) and cGVHD/Scl patients and will infuse FAP CAR Tregs to eliminate damaged cells. We will fill cGVHD pathophysiology knowledge gaps for mechanistic insights focused on T:B and Mac: fibroblast/myofibroblast/endothelial cell (fibrosis) crosstalk, test novel therapies in clinically relevant models and, with mature data, assist Dr. Pavletic to lead cGVHD CAR trials at the NIH Clinical Center using intramural funds

NIH Research Projects · FY 2026 · 2026-06

Project Summary/Abstract Epilepsy’s unpredictable seizures impair quality of life for >50 million people, and one-third of patients remain drug-resistant. Recent work from the Krook-Magnuson and Netoff labs demonstrates that cerebellar stimulation can truncate seizure duration. Additionally, studies from epilepsy-focused dynamical systems labs show that network instability linked to seizures can be revealed through perturbation-based measures, such as line length of evoked responses. Building on these advances, this predoctoral project pursues two independent yet complementary aims. Aim 1 will reduce behavioral seizure frequency by applying Gaussian-process/Bayesian optimization to identify cerebellar stimulation parameters that most effectively suppress behavioral seizure frequency in the ventral intrahippocampal kainate mouse model of temporal-lobe epilepsy. Aim 2 will improve seizure-risk assessment by delivering brief, sub-threshold hippocampal perturbations and quantifying evoked responses to assess “critical-slowing” and model proximity to seizure bifurcation, benchmarking performance against standard passive depth EEG based features. The integrated experimental–computational strategy is innovative in (i) shifting cerebellar neuromodulation from reactive seizure shortening to preventive seizure suppression, requiring optimization to succeed with fewer sampling data points, and (ii) coupling dynamical-systems theory with active probing to expose latent network fragility in real time. Expected outcomes include a stimulation procedure that lowers behavioral seizure incidence and a high-fidelity risk metric to measure seizure risk dynamically. Together they will advance the field toward anticipatory intervention. Training will take place through the Graduate Program in Neuroscience at the University of Minnesota, leveraging resources such as the Center for Neuroengineering and the broader university research infrastructure. Under the mentorship of Dr. Esther Krook-Magnuson (sponsor; epilepsy and circuit physiology) and Dr. Theoden Netoff (co-sponsor; optimization algorithms for neurostimulation), I will receive integrated training across neuroscience, engineering, and mathematics. This includes advanced coursework, hands-on experience with in vivo electrophysiology, and guided development in machine learning for neural data analysis. The fellowship will equip me to build the interdisciplinary expertise necessary to launch an independent research career focused on neuromodulation, predictive modeling, and translational epilepsy research.

NIH Research Projects · FY 2026 · 2026-06

Project Abstract This project focuses on the development of long-acting opioid antagonist, mixed agonist-antagonist, and agonist nanoparticle formulations for the treatment of overdose, reduction of opioid withdrawal, and prevention of renarcotization from fentanyl and fentanyl analogs (F/FAs), including carfentanil. These compounds are extremely potent mu opioid receptor (MOR) agonists that are easy to manufacture in clandestine laboratories and used as adulterants in counterfeit prescription medications and illicit street mixtures. Because of their widespread availability, F/FAs are responsible for over 70% of overdose deaths in the US. While antagonists like naloxone and nalmefene are FDA approved to treat opioid overdose, they can precipitate severe opioid withdrawal and may be insufficient to prevent toxicity and renarcotization in accidental F/FA overdoses. Renarcotization occurs when the antagonist effects wear off and the respiratory depressive effects of the opioid return due to differences in elimination half-lives. Management of overdoses involving F/FA is complex, often requiring intensive acute medical care and extended observation of patients. To reverse fentanyl-induced respiratory depression and prevent fentanyl effects out to 48 hours, our group has developed a novel long-acting naloxone nanoparticle (cNLX-NP). This project will focus on further development of this naloxone nanoparticle as well as development of other antagonist formulations (nalmefene and naltrexone), mixed agonist-antagonist (MAA) formulations (buprenorphine, nalbuphine, and butorphanol), and an agonist formulation (desmetramadol). The specific aims of the project are Aim 1: Develop antagonist nanoparticles, determine the lowest effective doses of each nanoparticle, and test whether antagonist nanoparticles induce anhedonia or precipitate opioid withdrawal via somatic signs; Aim 2: Develop MAA and agonist nanoparticles, determine the lowest effective doses, test whether MAA and agonist nanoparticles elicit rewarding, antinociceptive, or anhedonic effects, and test whether these nanoparticles precipitate opioid withdrawal; Aim 3: The 3 most effective formulations will be further characterized by determining the highest F/FA doses that each formulation is effective against, testing efficacy out to 120 hours, testing efficacy against a combination of fentanyl and alprazolam, test the efficacy in animals chronically exposed to fentanyl, characterize the pharmacokinetics of the lead formulation, and determine whether these formulations (which include a mixture with free naloxone) precipitate opioid withdrawal in rats using intracranial self-stimulation.

NIH Research Projects · FY 2026 · 2026-06

ABSTRACT Adrenoleukodystrophy (ALD), a rare X-linked neurometabolic disorder, occurs due to a defective transporter protein crucial for peroxisomal fatty acid oxidation. This results in the detrimental accumulation of very long- chain fatty acids (VLCFAs), a key contributor to nerve injury in ALD. This disorder encompasses diverse neurological phenotypes, and neither genotype nor biochemical features reliably predict its progression. The most severe manifestation is cerebral ALD, in which, without treatment, half of the affected boys succumb to the disease within five years of displaying clinical symptoms. With the nationwide integration of ALD into newborn screening, an increasing number of patients with an ALD variant are being identified, emphasizing the urgent need for safe and effective treatments to prevent disease progression. All individuals with this genetic defect, including females, will eventually develop progressive neurological symptoms, if left untreated. Presently, no approved treatment exists that can lower VLCFA levels in pre-symptomatic patients to prevent or mitigate the onset of cerebral disease. Our preliminary results demonstrate that nervonic acid, a monounsaturated fatty acid, can significantly decrease VLCFA accumulation in blood and tissues in mice. In this project, we propose to conduct well-designed, placebo-controlled study of nervonic acid as a preventive therapy for cerebral ALD (cALD) in a relevant mouse model to assess preclinical efficacy. We hypothesize that nervonic acid prevents or mitigates cerebral disease in ALD as indicated by reduced VLCFA accumulation and neuroinflammation. In Aim 1, we will evaluate the efficacy of nervonic acid in a cALD mouse model. Following administration of optimal doses, we will confirm the biochemical improvements and cerebral efficacy of nervonic acid and assess its effect on neuromotor and behavioral deficits observed in these mice. In Aim 2, we will characterize the exposure-response relationship of nervonic acid in mice. Using a combination of dose-escalation studies and long-term monitoring following dosing, we expect to determine the exposure (drug concentrations)-response (decrease in VLCFA-containing lipids) relationship of nervonic acid. This critical information will then guide the selection of appropriate dosage regimens for clinical studies. Impact: The findings from these nervonic acid efficacy studies hold vital clinical significance as a prospective therapeutic for the primary prevention of cALD and improve outcomes. Further, gaining a clear pharmacological understanding of NA will facilitate translation to human use and increase the likelihood of success by determining optimal dosage regimens for studies in adults and pre-symptomatic boys with ALD. Nervonic acid has the potential to be developed as a noninvasive, orally administered intervention suitable for all individuals carrying the genetic mutation.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY/ABSTRACT This project is fueled by candidate’s ambitions to build an independent academic research program at the intersection of biochemistry, physiology, and analytical chemistry, that explores intermediary lipid metabolism and shifts in subcellular lipid pools, to understand the molecular origins of metabolic diseases. This training project focuses on metabolic dysfunction-associated steatotic liver disease (MASLD) and steatohepatitis (MASH) which are the leading cause of liver disease in Western countries. Due to complex systemic and liver- autonomous factors influencing MASLD-MASH development and progression, treatment options are scarce, triggering an urgent need to better understand the molecular origins to develop new drug targets. The primary objective of this project is to investigate the role of ketone body-sourced polyunsaturated fatty acids (PUFAs) in the development of MASLD-MASH, implicating ketogenesis as a potential therapeutic target. This project will test the central hypothesis that ketogenesis-derived carbon sources malonyl-CoA to hepatic PUFA elongation, via acetoacetyl-CoA synthetase (AACS), which protects against MASLD-MASH by suppressing absolute rates of hepatic de novo lipogenesis. In Aim 1, a convergence of subcellular fractionation and mass spectrometry (MS)-driven proteomics and stable isotope tracing will be leveraged to study the co-localization of AACS and PUFA elongation enzymes within peri-lipid droplet mitochondria-associated membranes. In Aim 2, the hypothesis that ketone-sourced PUFAs protect against liver injury and inflammation will be tested using three loss-of-function mouse models, that strategically lack key ketone body metabolic enzymes, and a suite of physiological and analytical methodologies to quantify indices of liver metabolism and injury. These findings will deepen our understanding of the link between ketogenesis, PUFAs, and liver injury, and in so doing, implicate interventions such as intermittent fasting or low-carbohydrate diets that provoke ketosis as therapeutic options during MASLD-MASH. This project will take place in the Division of Molecular Medicine at the University of Minnesota Medical School. Molecular Medicine houses multiple labs focused on leveraging cutting-edge analytical tools to acquire high resolution molecular and phenotypical profiles, to better understand the metabolic underpinnings of human diseases. In this environment, and in conjunction with my co-sponsors, collaborators, and consultants, the outlined training plan will (1) provide proficiencies in subcellular fractionation, proteomics, and specialized training in shotgun lipidomics, and (2) provide experiences leveraging these tools to study in vivo diseases using dietary and genetic mouse models. Collectively, this will generate a unique and highly personalized training experience to prepare for a career focused on studying the molecular origins of metabolic diseases.

NIH Research Projects · FY 2026 · 2026-06

Bans on menthol as a characterizing flavor in combustible cigarettes have been implemented in multiple US jurisdictions with additional bans proposed. Naturalistic and experimental studies suggest that such bans are associated with decreased smoking in those who smoke menthol cigarettes but that a large proportion continue to smoke combustible products by switching to non-menthol cigarettes. A potentially less harmful pattern of tobacco use would be to switch to non-combustible products if such use decreased combustible cigarette smoking and did not undermine complete tobacco cessation. Among the available non-combustible tobacco products, nicotine pouches are a relatively new, rapidly growing and largely understudied product category that do not contain tobacco leaf and are neither aerosolized nor inhaled. On a continuum of risk, nicotine pouches are therefore likely among the least harmful tobacco products. Considering that people who smoke menthol cigarettes prefer menthol/mint flavored non-combustible products when switched to non- menthol cigarettes, it is important to assess how flavors in pouches affect the extent to which they are acceptable substitutes for menthol cigarettes. Using an experimental marketplace, this application aims to 1) Evaluate the acceptability of nicotine pouches relative to medicinal nicotine in people who smoke menthol cigarettes; 2) Evaluate how flavors influence the substitutability of nicotine pouches for non-menthol cigarettes; and 3) Evaluate the effects of a menthol cigarette ban on pouch use. The central hypotheses are that nicotine pouch uptake is greater than that of medicinal nicotine; that flavored pouches will lead to more substitution for cigarettes than unflavored pouches; and that a menthol cigarette ban will further increase the extent of pouch substitution. We propose a randomized study in which 252 people who smoke menthol cigarettes will for 6 weeks receive all tobacco products from experimental marketplaces designed for the study that will simulate either: 1) a ban on all pouch flavors; 2) a flavor ban that exempts menthol flavors in pouches; 3) no flavor ban (i.e., all flavors available) or 4) a control condition in which menthol cigarettes continue to be available along with all pouch flavors. In all conditions, non-menthol cigarettes and medicinal nicotine are available but menthol cigarettes are only available in the control condition. The study will provide data, under simulated real-world conditions, on the amount of each product used (including cigarette use), biomarkers of exposure, motivation to quit smoking, and subjective ratings of the products used. This work is innovative as there is little data regarding the impact of nicotine pouches and their flavors in the context of menthol removal in cigarettes. It is significant in that it assesses how these products are likely to be perceived and used by people who smoke thereby building a scientific basis for determining the public health impact of their availability, acknowledging that studies are also needed to assess the impact on youth. It is also significant in that it would contribute to the evidence that a menthol ban would accelerate reducing harm from smoking.

NIH Research Projects · FY 2026 · 2026-06

Project Summary/Abstract Urolithiasis (urinary stone disease) is a rapidly escalating public health crisis worldwide. The prevalence of kidney stones has more than doubled over the past 40 years, yet minimal new strategies for disease prevention have become available for decades. Calcium oxalate (CaOx) is one of the most common types of urolith, but this mineral composition requires expensive methods of physical removal. Following stone removal procedures, recurrence rates are as high as 50% in 5 years, highlighting an urgent need for novel, effective, and accessible stone prevention strategies. Disturbances in calcium balance constitute the most common metabolic abnormality underlying CaOx stone formation, yet the causes of these imbalances are often not identified. Recent discoveries suggest that the microbiome and microbial metabolites may influence stone formation. For instance, patients with CaOx urolithiasis exhibit deficiencies in short chain fatty acids (SCFAs), a class of metabolites produced in the gut following microbial fermentation of dietary and prebiotic fibers. In rodent models of urolithiasis, SCFA supplementation markedly reduces CaOx renal crystallization, potentially by lowering urinary oxalate and exerting anti-inflammatory effects in the kidneys. However, the relationship between SCFAs and calcium homeostasis in the context of CaOx urolithiasis remains poorly understood. This gap in knowledge is especially important given the high prevalence of idiopathic calcium disturbances in CaOx stone formers. In this study, we will use a multi-omics approach to delineate the interplay between the microbiome, SCFAs, calcium regulation, and stone risk in a natural canine model of CaOx urolithiasis. Dogs are a particularly powerful translational model for studying CaOx stone formation, as dogs form CaOx stones commonly and share several risk factors and features of disease pathology with humans. This includes parallels in calcium disturbances, microbiome abnormalities, and lower abundances of SCFA-producing bacteria. First, we will perform a prospective, case-control study to define interactions between SCFAs and individual calcium regulatory pathways in dogs with and without CaOx urolithiasis. This will bridge major gaps in our understanding of the microbiome and its role in stone pathogenesis. Second, we will examine how these pathways can be manipulated using prebiotic fibers to enrich SCFA production. This will provide practical insight into an inexpensive and accessible dietary supplement as a potential new stone prevention strategy. Third, we will determine how specific microbial and metabolic features predict stone recurrence. This will help us identify potential risk factors for stone recurrence that can be used to guide patient monitoring. We have assembled a powerful multidisciplinary mentoring team that is uniquely suited to guide this project. Data from this work will also lay the foundation for a future R01 funding application. With the resources and protected time provided through this award, I will advance my professional training and create a clear path towards independence as a translational microbiome and urological researcher.

NIH Research Projects · FY 2026 · 2026-06

ABSTRACT In recent years the 2AR adrenergic receptor (2AR) agonist xylazine has been incorporated in the illicit use of fentanyl, resulting in serious adverse pharmacological effects in users that are poorly understood. The synergistic interactions between mu-opioid receptors and 2AR adrenergic receptors have been long described. Additionally, the ameliorating effects that 2AR adrenergic receptor agonists exert on withdrawal from chronic exposure to mu-opioid receptor agonists is well-established. However, the interaction between xylazine and mu- opioid receptor agonists has been minimally investigated. We have previously extensively evaluated neuropharmacological interactions between clinically used 2AR adrenergic receptor agonists (e.g. clonidine, dexmedetomidine, brimonidine, and moxonidine) and opioid agonists ; we have also defined the mechanism of such interactions both in opioid-naïve rodents and those chronically exposed to opioids. We have also have established a model of fentanyl self-administration in mice. Applying these approaches to evaluation of the combination of xylazine with fentanyl will characterize the neuropharmacology of their interaction. The objective of this application is to test the combination of xylazine and fentanyl in pre-clinical models of analgesia, sedation, opioid tolerance, opioid withdrawal, and opioid self- administration. The central hypothesis of this application is that the pharmacology of the xylazine interaction with opioids is similar to that of the clinically used 2AR agonists. This proposal has two specific aims: i) Define the neuropharmacological mechanisms of xylazine when given alone and in combination with fentanyl. ii) Define the mechanism of the impact of xylazine on oral fentanyl self- administration. At the completion of our research, our expected outcomes are that we will have determined the neuropharmacological interactions of xylazine with fentanyl in well defined measures of analgesia, sedation, opioid analgesic tolerance and withdrawal and opioid self-administration. Our studies will have a positive impact because knowledge gained from pursuit of this program will significantly expand the current understanding of fentanyl and xylazine interactions, which is currently very limited. Such information will inform future approaches aimed at addressing harms associated with the introduction of xylazine into the illicit fentanyl use environment.

NIH Research Projects · FY 2026 · 2026-06

Project Summary There is an unmet need for the development of whole-brain level non-invasive metabolic neuroimaging tools for quantitatively and reliably detecting abnormal energy metabolism and impaired mitochondrial function in the early stage of the Alzheimer’s disease (AD) and brain tumors. Phosphorus-31 (31P) magnetic resonance spectroscopy imaging (MRSI) at ultra-high field strength (7 Tesla) is a non-invasive metabolic imaging method for investigating essential aspects of energy metabolism involving the adenosine triphosphate (ATP) synthesis and utilization in the human brain. The 8-channel 31P transceiver array coil offers better imaging sensitivity and SNR compared to conventional volume coils, but is limited by the inhomogeneous B1+ field of the RF array coil. We will develop a phase-only parallel transmission (ppTx) interface that allows continuously adjusting the RF phase for each transmit channel and B1+ phase shimming of the RF array, to significantly improve the B1+ distribution (∝ flip angle [FA]) of the array RF coil to enhance the FA uniformity and signal-to-noise ratio (SNR) of 31P MRSI across the whole brain. Using the proposed technique, we will quantify, for the first time, the molar concentrations of inorganic phosphate (Pi), phosphocreatine (PCr), ATP, and nicotinamide adenine dinucleotide (NAD), as well as determine the intracellular pH value throughout the brain, all of which are tightly involved in ATP metabolism within brain cells. The ppTx interface is digital-controlled, RF broadband, and portable on the MRI scanner's patient table, and thus can be applied to 1.5T, 3T, and 7T MRI scanners that do not have the parallel transmission (pTx) option.

NIH Research Projects · FY 2026 · 2026-06

I. PROJECT SUMMARY The abundance of available genome sequence information across the tree of life coupled to advances in DNA synthesis and genetic engineering tools have enabled innovative approaches to genome-guided natural product discovery1. Unfortunately, the success-rate of any individual approach (e.g., heterologous expression, regulator engineering, promoter replacement) is still low. Also, existing approaches are challenging to integrate into high- throughput natural product (NP) discovery pipelines. Each biosynthetic gene cluster targeted for activation/NP- discovery requires a bespoke set of genetic reagents (plasmids, sgRNAs, etc.) that must be introduced to the host genome using transgenesis pipelines that have not changed in the past thirty years. Put another way, plasmid creation and transgenesis protocols stand as major bottlenecks that prevent research groups from leveraging the abundant strain and genome sequence resources for high-throughput, genetics-enabled NP discovery. We propose a simple but radical shift in classical approaches by using universal genetic reagents and high- throughput transgenesis to invert the role of strain engineering in NP drug discovery campaigns. In this MIRA proposal we will elaborate on this approach, provide preliminary data to support its effectiveness, and describe plans to expand the technology from a proof-of-concept towards a new paradigm for NP drug discovery. The first innovation in our approach is a shift from bespoke genetic reagents towards universal ‘performance- enhancing plasmids’. These plasmids will encode the expression of transgenes designed to increase secondary metabolism in any recombinant Streptomyces host. Example transgenes include (i) conserved pleiotropic regulators, (ii) mutant RNA polymerases and ribosomal proteins, (iii) the bldA tRNA, (iv) pathways to overproduce precursor pools and/or posttranslational modifications to support secondary metabolism, and (v) combinations of the above elements. This circumvents a major bottleneck in genetic engineering. The second innovation is the development of a novel Streptomyces transgenesis pipeline that incorporates single-pot transgenesis, outgrowth, and NP discovery. By applying this approach in multi-well plates, we will be able to create and screen hundreds of transgenic Streptomyces strains (i.e., the same performance-enhancing plasmid in hundreds of unique genomic contexts). LC-MS/MS coupled with molecular networking will identify novel, over-produced NPs that will be structurally characterized from clonal cultures using traditional scale-up and analytical chemistry approaches. We have a solid foundation to begin work in scaling this approach for finding novel antiviral terpenoids from curated collections of actinobacteria. We will present plans to scale this in future years toward other biological activities (e.g., antibacterial, anticancer) and NP biosynthetic classes (e.g., nucleosides, aminoglycosides, polyketides).

NIH Research Projects · FY 2026 · 2026-06

ABSTRACT Prostate cancer is one of the most common malignancies and a leading cause of cancer-related deaths in American men. In advanced prostate cancer, androgen deprivation therapy (ADT) is the standard of care. While most patients initially respond to ADT, they will eventually develop castration resistant prostate cancer (CRPC). CRPC is a lethal and aggressive disease stage with limited treatment options. A novel therapeutic strategy for CRPC is bipolar androgen therapy (BAT). Clinical trials have demonstrated both safety and efficacy, as well as improvements in quality of life with the use of BAT. However, the patient populations best suited for BAT and the mechanism of anti-tumor activity in patients who respond to BAT remain undefined. CRPC is associated with amplification and overexpression of the androgen receptor (AR). One mechanism of AR amplification is extrachromosomal DNA (ecDNA). ecDNAs are Mb-sized circular structures located in the nucleus that allow for increased rates of transcription and replication, with new studies estimating that 1/3 of CRPC tumors likely contain AR ecDNA. Preliminary data in this project indicate that CRPC patient-derived xenografts (PDXs) classified as positive for AR ecDNA are sensitive to BAT. Based on this finding, my research training will explore AR ecDNA as a therapeutic vulnerability and biomarker to identify patient populations that should be considered for BAT. The central hypothesis is that AR ecDNA levels are dynamic during ADT and BAT, and that BAT inhibits AR ecDNA-positive CRPC by inducing R-loops, transcription-replication collisions, and DNA damage on ecDNA. Aim 1 will utilize computational tools to investigate the composition of ecDNA and how treatment affects quantity of AR ecDNA structures. Aim 2 will investigate the mechanism of BAT in ecDNA- positive tumors, assessing the vulnerability of these structures to excess DNA damage during treatment. This project is specifically designed to provide rigorous training in key areas for my development as an independent physician-scientist. Aim 1 will provide hands-on training in computational biology and who genome sequencing data. Aim 2 will build my expertise in advanced molecular biology techniques to probe mechanisms by which BAT enhances transcription and DNA damage. Completing this project will advance our understanding of the role of AR ecDNA in BAT sensitivity and biological mechanisms of action and provide me with the conceptual knowledge and technical skills necessary to lead a research project focused on translating mechanistic discoveries into novel cancer therapies.

NIH Research Projects · FY 2026 · 2026-06

Abstract. Dental plaque is among the most complex microbial communities in humans, second only to the gastrointestinal tract. During community development, bacteria interact with and respond transcriptionally to proximal bacteria and host factors such as MUC5B. In this context, Streptococcus gordonii is a key model pioneer colonizer, transcriptionally discriminating MUC5B from a low-density salivary protein fraction using an outside-in signaling (OIS) circuit and also responding to Actinomyces naeslundii and other early plaque colonizers during coaggregation. We will learn whether OIS recognizes and responds to A. naeslundii polysaccharides and MUC5B and both together. The OIS circuit is comprised of recognition adhesin proteins known as SspA and SspB, lipoteichoic acid (LTA) for signaling across the cell wall into the membrane, and an intramembrane two-component system called SraSR for transcriptional regulation of adhesins sspA and sspB, tagatose pathway genes, and luxS (quorum sensing). Intramembrane C-terminal peptides (C-peps) cleaved from LPXTG-motif surface proteins by sortase A (SrtA) also modulate SraSR transcription, including adhesins as part of a larger transcriptional profile. Gaps to be addressed: S. gordonii interacts with some plaque species and competes with others, yet the vital role that pioneer S. gordonii plays in shaping the biofilm community and structure remains largely unknown. We seek to understand how the adhesin genes in the transcriptional responses of S. gordonii to other early plaque colonizers help shape the initial plaque community. Specifically, we aim to: 1. Examine the mechanism that S. gordonii uses to respond transcriptionally during co-aggregation with A. naeslundii. 2. Establish the structural basis and outcomes of interactions between the S. gordonii SspB adhesin V-region and A. naeslundii polysaccharides. 3. Characterize how OIS signaling influences the assembly of a synthetic oral bacterial community. Community changes in response to stressors may affect plaque composition and biogeography and underlie the transition from health to significant diseases of the oral cavity ranging from dental caries and periodontitis to squamous cell carcinoma. Given the burden and concomitant costs of disease that occur with unchecked changes in the composition of oral biofilms, our proposed research project will provide a better understanding of fundamental adhesive mechanisms that direct the membership and architecture of dental plaque and suggest novel targets for potential therapeutic interventions.

NIH Research Projects · FY 2026 · 2026-05

The afferent innervation of the kidney remains poorly understood. Gaps in knowledge persist regarding the precise location of afferent renal nerves (ARN) in the kidney, their origin in spinal and vagal sensory ganglia, their interaction with central nervous system (CNS) interoceptive circuits, and their functional properties under physiological and pathophysiological conditions. Clinical trials for catheter-based renal denervation for drug resistant hypertension (HTN) have shown additional benefits independent of reducing blood pressure. These benefits can only be explained by ARN ablation. Findings from our preliminary data and the literature suggest that input to the CNS from TRPV1-expressing kidney-innervating primary afferent neurons (TRPV1+ kiPAN) are involved in the central regulation of blood pressure. The objective of this proposal is to define the diversity of ARN that express the capsaicin receptor TRPV1 and the impact of their CNS input on central interoceptive networks. Our central hypothesis is that input from distinct subtypes of TRPV1+ kiPAN can modulate CNS interoceptive circuits. This proposal will address this hypothesis in two aims. In Aim 1, we will define the modulation of central interoceptive circuits by TRPV1+ ARN under normal conditions and during neurogenic HTN. We hypothesize that increased activity of TRPV1+ ARN is reflected in altered neuronal activity in central interoceptive circuits under normal and HTN conditions. We will use the TRAP2 transgenic mouse line to conduct global analysis of CNS neuronal activation after capsaicin stimulation of ARN to identify CNS regions that process TRPV1+ ARN input. We will use fluorescent in situ hybridization (FISH) to determine the neurochemical subtype of these activated neurons and employ viral vector tracing to define the projections of brainstem and spinal neurons that receive direct input from TRPV1+ ARN. In Aim 2, we will determine if subtypes of TRPV1+ kiPAN have distinct contributions to renal interoception. We will use single-cell RNA sequencing, FISH, and neurotracing to test the hypothesis that distinct TRPV1+ kiPAN subtypes innervate the renal cortex and pelvis, and that these subtypes undergo specific transcriptional changes during neurogenic HTN. We will test if TRPV1+ kiPAN are spontaneously active and if their activity increases during neurogenic HTN using in vivo imaging of identified TRPV1+ kiPAN, and use FISH to identify the transcriptional subtypes. We will visualize the peripheral processes of TRPV1+ kiPAN within the renal pelvis and cortex using retrograde viral vector tracing to test if the kidney is innervated by spinal and vagal primary afferent neurons with different peripheral termination patterns. There is a critical need for thorough analysis of the interoceptive functions of ARN in order to harness the therapeutic potential of their manipulation through pharmacological or non-invasive neuromodulation approaches. The proposed work will transform our understanding of the diversity of kiPAN and the integration of their inputs in central interoceptive circuits. We expect that these studies will provide insights into central and peripheral neuroplasticity during neurogenic hypertension, which may inform future therapeutic development.

NIH Research Projects · FY 2026 · 2026-05

Project Summary/Abstract Deciphering how gene perturbations reshape cellular signaling networks is essential for un- derstanding disease mechanisms and discovering therapeutic targets. However, current pooled CRISPR screening platforms lack the scalability, throughput, and protein-level resolution needed to capture complex cellular responses, especially those governed by post-translational modifications. This collaborative project between Dr. Xiaokang Lun’s lab at the University of Minnesota and Dr. Jellert Gaublomme’s lab at Columbia University seeks to overcome these limitations by developing CRISPRmap-CyTOF, a novel single-cell screening platform that integrates high-complexity pooled CRISPR perturbations with high-dimensional protein-level profiling using mass cytometry. The platform builds on the CRISPRmap combinatorial barcoding strategy (developed by the Gaublomme lab) and a photocrosslinking-based DNA stabilization method (developed by the Lun lab), combining mass cytometry-based signaling network analysis to simultaneously decode thousands of gene perturbations and quantify ∼30 protein or phospho-epitope markers in individual cells. This one-step, high-throughput readout enables proteome-resolved CRISPR screens at single-cell resolution across millions of cells. In Aim 1, we will re-engineer CRISPRmap for compatibility with mass cytometry, optimizing probe chemistry, staining conditions, and barcode decoding algorithms to enable high-fidelity linkage between gene perturbation and signaling phenotypes. In Aim 2, we will apply CRISPRmap-CyTOF to systematically profile early and late signaling responses to a kinome-scale CRISPR knockout library in chronic myeloid leukemia (CML) cells, with and without tyrosine kinase inhibitor (TKI) treatment. These experiments will identify kinases that drive CML progression, uncover novel TKI resistance mechanisms, and generate integrative signaling biomarkers predictive of disease outcome. Together, this work will deliver a transformative technology for functional genomics and a mecha- nistic framework for dissecting therapy resistance in cancer.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY Measles virus (MeV) is a highly contagious pathogen that causes significant morbidity worldwide. Despite the availability of an effective vaccine, MeV continues to cause outbreaks, highlighting the need for alternative treatments that target viral replication and pathogenesis. The MeV polymerase complex, consisting of the large protein L and phosphoprotein P, plays a critical role in RNA transcription and replication, making it a promising target for antiviral strategies. However, key aspects of the complex’s structure and function remain poorly understood, including how it interacts with RNA, how it distinguishes between transcription and replication, how the L and P proteins work together during RNA synthesis, and how the polymerase complex cooperates with the nucleoprotein N during RNA replication. Additionally, the role of other factors in regulating polymerase activity has yet to be well defined. Our overall objectives for the proposed funding period are to (1) determine the structural and functional properties of MeV polymerase in its various functional states, including initiation, elongation, and termination; (2) explore the dynamics of the polymerase-nucleocapsid interaction; and (3) investigate the role of the viral co-factor C protein in regulating RNA synthesis. We hypothesize that understanding these interactions will reveal insights into the mechanisms of how MeV polymerase functions and how its regulation can be targeted for therapeutic intervention. To address these questions, we will utilize a combination of cryo-electron microscopy (cryo-EM), protein-RNA biochemistry, and viral minigenome systems. The proposed work aims to provide new insights into the structural and functional mechanisms governing MeV replication, with the potential to identify new targets for antiviral therapies that could help control MeV outbreaks.

NIH Research Projects · FY 2026 · 2026-05

ABSTRACT The long-term goal of our research is to determine how opioid exposure modifies nucleus accumbens microcircuits, and use this knowledge to reverse or prevent maladaptive forms of opioid-evoked plasticity. This proposal focuses on a specific type of nucleus accumbens neuron, the low-threshold spiking interneuron (LTSI), which express unique molecular markers not present in other nucleus accumbens cell types (e.g., somatostatin and neuronal nitric oxide synthase). LTSIs are critical gatekeepers of nucleus accumbens output, and prior research shows that LTSIs regulate the reinstatement of drug-seeking behavior. We recently identified LTSIs as a novel target of mu opioid receptor (MOR) action in the nucleus accumbens. This evidence and our preliminary data suggest that nucleus accumbens LTSIs may be modulated by, and in turn regulate, opioid seeking/taking. The specific goal of this proposal is to study interactions between LTSIs and other nucleus accumbens cell types during the acquisition, extinction, and reinstatement of intravenous fentanyl self-administration in mice. Our central hypothesis is that fentanyl self-administration causes adaptations in LTSI function that result in vulnerability to reinstatement of fentanyl-seeking behavior. We will test this hypothesis across three specific aims. AIM 1 is to measure LTSI synaptic output after acquisition of fentanyl self-administration, and determine the contribution of MOR expression. We will measure synaptic transmission ex vivo after fentanyl self-administration in vivo, using transgenic mice to optogenetically stimulate LTSIs and record the response of post-synaptic neurons in acute brain slices. We will also combine viral vectors and transgenic mice in an intersectional genetic strategy to selectively delete MOR expression from nucleus accumbens LTSIs, and then rescue MOR signaling using optogenetics. AIM 2 is to monitor dynamic interactions between LTSIs and other cell types throughout fentanyl self-administration. Using fiber photometry, we will simultaneously monitor the dynamic activity of LTSIs and other nucleus accumbens cell types. These signals will be recorded longitudinally as mice acquire, extinguish, and reinstate fentanyl taking/seeking. AIM 3 is to manipulate LTSI regulation of other cell types during reinstatement of fentanyl-seeking. Following acquisition and extinction of fentanyl self-administration, we will perform chemogenetic stimulation of LTSIs, with or without genetic block of GABA release. We will then perform chemogenetic stimulation of LTSIs and concurrently activate or inhibit other nucleus accumbens cell types, to determine if chemogenetic activation of LTSIs produces reinstatement via regulation of downstream neural activity. Successful completion of these experiments will yield new information about the fundamental operation of LTSIs within nucleus accumbens microcircuits. Our results may also highlight LTSIs as a new therapeutic target for decreasing the reinforcing properties of fentanyl and reducing vulnerability to reinstatement/relapse.

NIH Research Projects · FY 2026 · 2026-05

ABSTRACT The University of Minnesota (UMN) Innovative Approaches to Trauma Care Program will provide dual mentorship-based training to surgery residents, critical care fellows, and emergency medicine fellows in a broad variety of topic areas that relate to trauma and critical care. Trauma is the number one cause of death for people under the age of 46, and deaths are more likely to occur in less severely injured patients in rural settings compared to their urban counterparts. Major gaps and pressing questions in the area of optimal trauma care include 1) evaluating the effect of prehospital care interventions on patient-centered outcomes; 2) optimizing trauma/critically ill patient monitoring (e.g., wearable sensors, AI-assisted early warning systems, and prehospital telemedicine); 3) advancing innovative, patient-centric clinical trials targeting the leading causes of death and disability post-injury; and 4) developing novel solutions to improve rural trauma care. Physician scientists are uniquely positioned to create and lead interdisciplinary teams due to their broad network of health professionals and non-physician scientists. However, physician scientists are becoming exceedingly rare, with only 1.5% of active physicians currently funded by the NIH1. Our program will provide a research focused curriculum, access to high-quality mentors and collaborators, and provision of protected time, all critical to launching early career physician scientists. Curriculum: Our two-year program will consist of four tracks including 1) Clinical Trial Innovation, 2) Rural Trauma Care, 3) Trauma and Critical Care Outcomes, and 4) Translational Preclinical Research. The trainee project topic areas will broadly include trauma care, including: early resuscitation, wound infection and prevention, hemorrhagic shock, and prehospital interventions after severe injury. By the end of the fellowship, trainees will also receive a relevant Master's degree. Co-Directors, Mentors, and Collaborators: The Co-Directors of this program are physician scientists in Surgery and Emergency Medicine and leaders of the Translational Center for Resuscitative Trauma Care, a Center at the University of Minnesota Medical School that improves trauma care through the creation of technologies and knowledge products deployable by first responders and hospital-based healthcare workers. Mentorship within our program also features nationally and internationally recognized mentors in Acute Care Surgery, Emergency Medicine, Prehospital Emergency Care, Critical Care, and Biostatistics. Moreover, we have unique training opportunities related to rural trauma care (University of Minnesota-Duluth and Minnesota Department of Health), established linkage of prehospital data to outcomes for rapid hypothesis development and testing (eso and UMN-led database), and innovative emergency care clinical trial design development and leadership (SIREN network and UMN School of Public Health). We have strong collaborations with four Minnesota Level 1 trauma Centers with a combined total of more than 15,000 trauma admissions/year with active clinical trial activity that can be leveraged by our trainees.

NIH Research Projects · FY 2026 · 2026-05

Project Summary/Abstract Cholangiocarcinoma (CCA) is an aggressive tumor with the urgent need of better therapeutical approaches. Our research has established that primary cilia, critical sensory organelles in cholangiocytes, are frequently lost or malformed in CCA, contributing to disease progression. Our findings also show that dysfunctional or experimental elimination of cilia transforms a normal cell into a hyperproliferative one. We also found that pathologic overexpression of SIRT1 leads to ciliary loss. Importantly, our preliminary data indicate that butyrate, a gut microbiota-derived short-chain fatty acid (SCFA) and a known SIRT1 inhibitor, restores ciliary expression and reduces tumor cell proliferation. Dietary fibers are indigestible polysaccharide digested by bacterial fermentation in the gastrointestinal tract until arrival to the colon, resulting in the formation of SCFAs including butyrate. Thus, presence of fiber diet and specific bacterial type may favor the production of the SCFAs, leading to different changes in the gut and distal organs. Our long-term goals are to understand the pathogenesis of the cholangiociliopathies and to develop new therapies for its treatment. Our overall objective for this proposal is to explore the novel hypothesis that dietary interventions that promote bacterial butyrate formation in the gut maintain and/or rescue ciliary expression in cholangiocyte to decrease disease progression. Our rationale is based on our previous findings and preliminary in vitro data showing that; (i) experimental deciliation of cholangiocytes induces cell proliferation and migration; (ii) overexpression of deacetylases, including SIRT1, are responsible for decreased ciliary expression; (iii) SIRT1 inhibition rescues primary cilia expression; and (iv) Butyrate inhibits SIRT1 activity and promotes ciliary expression decreasing cell proliferation. Thus, we propose here the novel CENTRAL HYPOTHESES that increasing butyrate levels in the gut (through fiber supplementation and/or probiotic microbiota modulation) will restore cholangiocyte primary cilia and reprogram tumor cell behavior, thereby slowing CCA progression. To test these central hypotheses, we propose two Specific Aims. In Aim 1 (Therapeutic Aim), we test the hypothesis that butyrate produced via fiber fermentation by gut bacteria, or delivered as oral tributyrin, can reduce CCA tumor growth and progression by promoting cilia-dependent tumor cell cycle arrest and cell death in rodent models of CCA. In Aim 2 (Mechanistic Aim) we Determine the mechanisms by which SCFAs restore primary cilia and alter tumor cell behavior, independent of Aim 1. Impact: These experiments will uncover novel and generalizable information on the fundamental mechanisms of therapeutic ciliary-restoration by the gut microbiota and dietary manipulations for CCA that may be applicable to other tumors and ciliopathies and have translational potential to improve outcomes for patients.

NIH Research Projects · FY 2026 · 2026-05

Project Summary Selective therapeutic delivery enables precision medicine. Overexpressed biomarkers provide delivery targets, yet moderate expression in healthy tissue results in on-target, off-disease toxicity thereby limiting the therapeutic window. Exquisite cellular targeting remains a challenge. Strategies to increase disease-specific delivery are sorely needed. To increase cell specificity, we will [1] advance from targeting biomarker presence to targeting high biomarker density, and [2] require the presence of two distinct biomarkers rather than one to reduce on- target / off-disease opportunities. These stringency factors will be achieved through low-affinity, high-avidity, cooperative binding via controlled heterobivalency (i.e., dual density-dependent molecular AND gate binders). While density-dependent binding has been achieved in several cases, more rigorous and systematic evaluation is needed to elucidate design principles and empower precise and tunable control to maximize selectivity. Thus, increased insight and efficient modular engineering will be especially valuable in rationally tailoring the engineered avidity and cooperativity in bispecific systems that are needed to achieve ultraselectivity. We pursue an innovative strategy that leverages modular synthetic miniproteins, systematic engineering, and assembly of molecular components through an integrated experimental/computational approach. Objective: Engineer targeting ligands that selectively bind cells expressing high levels of two antigens — i.e. dual density- dependent molecular AND gates — via engineered bispecific avidity. We pursue [1] specific application of bispecificity to engineer a Trop2/EGFR density-dependent bispecific targeting agent, with physiological validation in a murine tumor model; [2] broader elucidation of the molecular factors — both ligand and target — that dictate performance; and [3] establishment of a generalizable engineering platform for efficient generation of dual density-dependent molecular AND gates. Aim 1: Engineer dual density-dependent Trop2/EGFR AND gate via bispecific avidity. We will engineer a bispecific density-dependent binder to Trop2 and EGFR by modulating affinity, valency, and epitope, and the length, rigidity, and orientation of the linker. The benchmark for success is a ligand with a sub-nM EC50 for binding cells with dual high expression and ≥100:1 binding differential relative to cells with single-high density or dual-moderate density. Physiological targeting will be evaluated in murine xenograft tumor models via PET imaging and excised tissue gamma counting. Aim 2: Elucidate factors that drive dual density dependence in bispecific molecules. We will more broadly evaluate binding with a set of systematically varying ligands and receptors experimentally via flow cytometry and computationally via a mechanistic mathematical model. The impact of molecular design parameters and target settings on dual density dependence and binding selectivity will be quantified across multiple biomarkers for generality. Resolution of model mechanisms with experimental data will elucidate the factors that drive expression dependence and empower rational design of molecules with tailored expression dependence.

NIH Research Projects · FY 2026 · 2026-05

ABSTRACT Biomarkers of exposure and biological effects are invaluable tools for studying the etiology of environmentally- induced cancers, identifying individuals and populations at risk, and developing and evaluating preventive interventions. This proposal will leverage the expertise and the teaching and capacity building experience of the Masonic Cancer Center (MCC), University of Minnesota faculty to provide an innovative, hands-on course to develop skills necessary for employing biomarkers of carcinogen exposure and biological effects in studies of cancer etiology and prevention. The course is targeted towards researchers-in-training (RITs) such as predoctoral and medical students and postdoctoral trainees from the US and from countries that represent high burden of exposure to environmental and lifestyle cancer risk factors. We will recruit 10 RITs each year, with each cohort consisting of four US-based participants and – leveraging our existing partnerships – six participants from India, Thailand, and African countries (two participants from each site). Specific Aims are (1) Provide a course that offers structured didactic content and hands-on experiences for developing skills in biomarker measurement and data analysis and interpretation. The course will include virtual modules focused on the fundamentals of chemical carcinogenesis and examples of biomarker development and applications, and a 5- week in-person laboratory training in state-of-the art biomarker measurement techniques. (2) Provide course participants with an understanding of clinical and translational biomarker applications to inform cancer prevention and control. Participants will learn and practice methodologies for designing, planning, and implementing human cross-sectional and intervention studies employing biomarkers. This will include modules on the ethics of human subject research as well as the logistics of project management, such as participant recruitment, retention, compliance with study procedures, and biological sample collection, processing, and shipment. (3) Provide course participants with skills and opportunities to establish global collaborative partnerships for conducting biomarker-based research in the area of cancer prevention and control. Each participant will partner with a participant from a different country to develop a joint research proposal employing biomarker assessments. The proposal will address a cancer risk factor relevant to both countries (e.g., air pollution, tobacco, diet) and will be designed to ensure that its findings will serve to inform each country’s cancer prevention and control efforts. Our long-term goal is to establish a community of highly skilled researchers who can engage in collaborative, translational biomarker research aimed to inform policies and practices to reduce the global burden of cancer.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY Stable transmission of chromatin states, which encode epigenetic information, is essential for preserving gene expression patterns and maintaining cell identity across generations. Despite its importance, the mechanisms ensuring the intact transmission of chromatin structure and histone modifications during cell division remain poorly understood. The PI’s long-term goal is to elucidate the mechanisms governing epigenetic inheritance, with a primary focus on nucleosome dynamics. The specific objectives for this funding period are to: (1) determine the regulatory mechanisms governing parental histone transfer to newly replicated DNA strands during DNA replication; (2) investigate how parental histone transfer pathways influence replication fork recovery under replication stress; and (3) examine the DNA replication strand preference of chaperones for newly synthesized histone in epigenetic inheritance. His central hypothesis is that proper control of parental histone transfer safeguards the stability of chromatin status inheritance, and that the transfer process is highly regulated by a variety of mechanisms, including the iron-sulfur assembly pathway. In addition, he hypothesizes that the chaperones for newly synthesized and parental histone must cooperate to maintain chromatin state during replication. The proposed work leverages innovative methodologies, including advancements in the eSPAN technique (enrichment and sequencing of protein-associated nascent DNA). This technique, central to the project, enables precise mapping of protein interactions with the leading or lagging strands of replicating DNA. This technology allows researchers to interrogate the mechanisms regulating nucleosome dynamics during chromatin replication, an area that was previously difficult to study due to technical challenges. The outcome of this work will be elucidation of the regulatory mechanisms responsible for the transfer of parental histone H3-H4 tetramers and a better understanding of the importance of this process in chromatin stability. Aberrant regulation of proteins involved in chromatin replication, including histone chaperones, histone modification enzymes, and chromatin remodeling complexes, have been directly linked to breast, gastrointestinal, and prostate cancers. Thus, the mechanisms the PI uncovers for DNA replication–coupled histone transfer and assembly will not only provide new insights into the fundamental process of epigenetic inheritance, but also create new potential targets for human cancer therapies.

NIH Research Projects · FY 2026 · 2026-05

ABSTRACT Cancer survivors experience multiple age-related diseases that occur at earlier age than in cancer-free persons. Thus, cancer survivors age faster than those without cancer, and the accelerated aging is driven by prolonged cellular senescence. The detrimental role of senescence is mainly explained by the proteins that comprise senescent-associated phenotype (SASP). These SASP proteins may be released into blood. The central hypothesis of our proposed R21, responsive to PAR-23-255, is that, in cancer survivors, circulating post- diagnosis SASP proteins are associated with an increased risk of mortality, increased risk of cardiovascular death (CVD), and greater changes in frailty and kidney function. To assess circulating proteins after cancer diagnosis, we will use proteomic data measured by highly sensitive SomaScan assay in a large population- based cohort study – Atherosclerosis Risk in the Community (ARIC) of white and black participants. Using ~5000 SomaScan proteins measured three times over 20 years, we will create cross-sectional (Aim 1) and longitudinal SASP indexes (Aim 2) that will include 133 functional, biologically meaningful proteins that were identified in the published atlas of soluble SASP proteins. The unique feature of our novel longitudinal SASP index is that it will combine the characteristics of the cross-sectional index and rate of change in each SASP protein, and may better reflect the aging process than the cross-sectional metrics. The indexes will be created in the training set of cancer-free participants, internally validated in the remaining cancer-free participants, and then applied to cancer survivors to examine associations with all-cause mortality including mortality from cancer and from other causes, as well as the CVD risk and changes in frailty and kidney function. We will also explore whether associations of SASP indexes with outcomes differ in those with cancer and in cancer-free participants. Our multi-disciplinary study team is well-prepared to lead this work, with complementary expertise in cancer and molecular epidemiology, cancer survivorship and oncology, chronic disease, aging biomarkers, biostatistics, and proteomics analysis. The use of existing well-characterized ARIC data will enable a quick and cost-efficient testing of our hypothesis. The proposed study will have a significant impact because SASP indexes are biologically meaningful, and understanding their role in cancer survivors will not only inform risk-stratified cancer care and surveillance for age-related diseases, but may also serve as a potential target for anti-senescence drugs that are currently in clinical trials or under development.

NIH Research Projects · FY 2026 · 2026-04

ABSTRACT Bacteria shield themselves from the exterior environment with a rigid cell wall. The major structural component of this exoskeleton is the mesh-like peptidoglycan (PG), a heteropolymer composed of glycan strands and crosslinked stem peptides. Glycan strand polymerization and transpeptidation are catalyzed by penicillin-binding proteins (PBPs). Organisms express multiple PBP isoforms with varied spatiotemporal activity. Transpeptidation is inhibited by the b-lactams, a family of covalent inhibitors that mimic the terminal dipeptide structure of the nascent PG monomer; PBP inhibition results in growth abnormalities and cell lysis. Despite the therapeutic successes of b-lactam antibiotics, the current antimicrobial resistance crisis underscores the importance of understanding of PBP activity and inhibition, and cell wall composition more broadly. The Carlson Group studies PBP activity with chemical probes; however, achieving isoform selectivity is difficult due to spatial conservation of the transpeptidase active site. Specifically, the field lacks a comprehensive description of how conserved motifs in this active site mediate substrate and inhibitor binding. There is also a lack of understanding of how changes in the PBP activity profile alter PG composition. While nonuniformity of PG composition has been demonstrated in the context of bacterial resistance and host immune response to infection, a lack of bioinformatic methods for the unbiased identification of PG fragments impedes analyses of cell wall digests using liquid chromatography coupled to mass spectrometry (LC-MS). To address these outstanding questions in PBP activity and cell wall maintenance, the proposed research will use an alanine scan to map and quantify the contribution of conserved active site motifs in PBP2x from the Gram-positive pathogen Streptococcus pneumoniae to native substrate processing and b-lactam inhibition. Following initial assessment with a commercially available probe, this approach will be extended to a chemically diverse suite of b-lactams in live cell, in vitro, and in silico experiments to clarify how the active site permits occupancy of multiple inhibitor classes. Additionally, this project will use activity-based protein profiling and LC- MS/MS analyses to investigate how conditions mimicking the acidic infection microenvironment perturb PBP activity and PG composition. This work will be coupled to experiments probing the effects of transpeptidase inhibition and PBP deletion on PG structural diversity. Bioinformatic analyses of LC-MS/MS data will serve as crucial validation of the application of -omics strategies to interrogate PG composition. Ultimately, these aims will provide insight into key mediators of cell wall synthesis and maintenance, inform understanding of how bacteria leverage cell wall perturbations as defense mechanisms to evade antimicrobial threats, and inspire future probe design. The training plan will facilitate instruction in chemical biology, computational biochemistry, and bioinformatic -omics analyses through education in the group of Erin Carlson, and through collaborations at the University of Minnesota.

NIH Research Projects · FY 2026 · 2026-04

PROJECT SUMMARY Immunotherapies such as bispecific T-cell engagers (BiTEs) and chimeric antigen receptor (CAR) T cells have shown significant promise in treating hematologic malignancies. However, therapeutic responses vary between patients and across the different subtypes of leukemia and lymphoma. A key challenge in optimizing these therapies is the lack of in vivo preclinical models that accurately reflect both the patient's immune and cancer cell biology. Current patient-derived xenograft (PDX) models lack functional immune systems, while humanized mouse models typically involve healthy donor immune cells paired with cancer cells from a different donor. These immunologically mismatched, or allogeneic, models fail to replicate autologous immune-cancer cell dynamics and the effect the cancer microenvironment and therapy have on immune cell function. To overcome these limitations, we aim to develop innovative autologous humanized PDX models using leukemia and immune cells derived from the same patient. We will collect paired bone marrow (BM) samples from pre-B acute lymphoblastic leukemia (B-ALL) patients at diagnosis and remission. Hematopoietic stem and progenitor cells (HSPCs) from the remission BM will be expanded and transplanted into immunodeficient mice to generate humanized mice with intact immune systems. These mice will then be engrafted with diagnostic leukemia cells, creating PDX models with autologous immune and leukemia cells. In parallel, a second cohort of models will be developed using autologous peripheral blood mononuclear cells (PBMCs) and leukemia cells from the same patient. We hypothesize that these fully patient-derived autologous models will provide deeper insights into immune-leukemia interactions and enhance preclinical testing of immunotherapies. To test this hypothesis, we will pursue three specific aims. First, we will establish autologous PBMC- and HSPC- humanized models using samples from B-ALL patients with diverse genetic and risk subtypes. We will evaluate disease progression and immune responses longitudinally in these models. Second, we will assess the efficacy of a CD3/CD19 BiTE and CD19 CAR-T cells in these autologous models. Third, we will analyze the immunophenotypic and transcriptional profiles of cells from autologous models, allogeneic models, and patient samples before and after immunotherapy treatment to validate the translational relevance of the models. These studies aim to create robust, patient-specific models that can be used to test and optimize immunotherapies, ultimately improving their clinical impact in the treatment of hematologic malignancies.

NIH Research Projects · FY 2026 · 2026-04

Since its introduction, MRI has been a transformative tool in medicine and biomedical research. Beyond its remarkable contributions to imaging advancements, significant national resources have recently been directed toward utilizing MRI to investigate neuropsychiatric disorders in the human brain. Two notable initiatives in this area are the Human Connectome Project (HCP) and the BRAIN Initiative. The primary goal of the HCP is to map the functional and structural connectivity of the entire human brain—using functional MRI (fMRI) and diffusion MRI (dMRI), respectively—at the highest possible resolution. Meanwhile, the BRAIN Initiative focuses on developing cutting-edge technologies and methodologies to drive neuroscience discoveries. Theoretical predictions consistently highlight higher field strengths, particularly ultrahigh field (UHF, 7T and above) MRI scanners, as essential for achieving these ambitious milestones. In this context, the 10.5T scanner at CMRR has revolutionized UHF MRI, demonstrating the immense potential of UHF scanners to push the boundaries of imaging technology. However, despite its promise, the current radiofrequency (RF) and gradient technologies at 10.5T remain suboptimal, limiting the scanner’s ability to reach its full potential. Motivated by the transformative opportunities presented by the 10.5T scanner, particularly in neuroimaging, and leveraging our extensive expertise, this proposal seeks to overcome these technological barriers through innovative solutions targeting ultrahigh-resolution fMRI and dMRI. Our approach focuses on three key aims. Aim 1 seeks to advance RF technology to achieve the theoretically promised signal-to-noise ratio (SNR) while addressing RF excitation challenges at 10.5T. This will involve developing an innovative 80-element, shielded, tight-fitting hybrid array coil that combines loop-dipole designs with high-permittivity materials. The coil will be rigorously characterized for SNR, parallel imaging, and RF shimming performance. Aim 2 aims to develop a novel RF coil safety assessment technique to reduce overly conservative safety factors, thereby increasing input power limits while ensuring patient safety. This includes designing a patient-specific, physics-guided deep learning-based method to predict the safety parameters during scans. Aim 3 focuses on achieving mesoscopic-scale human fMRI and dMRI with unprecedented resolutions, including isotropic resolutions of 0.46 mm and 0.63 mm for whole-brain functional and structural connectivity mapping, respectively, and 0.27 mm for partial-brain fMRI studies. By addressing these critical technological challenges in RF coil design, safety assessments, and imaging protocols, this proposal aims to achieve imaging resolutions that surpass those currently available at 3T and 7T. These innovations will set a new benchmark in neuroimaging, advancing the goals of two major national initiatives—the HCP and the BRAIN Initiative—while unlocking new insights into brain structure and function.