University Of California, San Francisco

NIH Research Projects · FY 2026 · 2024-05

PROJECT SUMMARY Despite evidence that half of all surgical site infections (SSIs) may be preventable, SSIs continue to increase in the United States and are a substantial cause of morbidity, mortality, and healthcare costs. There is a lack of evidence-based guidelines for pediatric SSI prevention. Previous efforts to identify pediatric risk factors to inform actionable recommendations have been limited by small sample sizes and data availability. There is an urgent need to provide clinicians with evidence-based, individualized SSI risk and prevention recommendations to optimize patient care, reduce infection risk, and improve shared decision making and informed consent for children and families undergoing surgery. The objective of this proposal is to harness the power of machine learning to generate SSI risk prediction models using electronic health record (EHR) data to inform pediatric SSI care and the design of an EHR-based clinical decision support tool. This study will leverage a large national pediatric surgical dataset to train, validate, and test statistical and machine learning algorithms that will then be applied to an external test set from Stanford Medicine Children’s Health to evaluate performance and applicability for real-world clinical care (Aim 1). The investigators will then apply human-centered design to create and test the usability of an EHR-embedded clinical decision support tool prototype that provides clinicians with real-time, evidence-based SSI risk estimations and prevention guidance (Aim 2). The long-term goal of this project is to produce a clinical decision support tool that will be ready for prospective testing to augment real-time SSI prevention decision making to help clinicians care for surgical patients with higher reliability using evidence-based, patient-specific actions. This research will support NICHD’s focus on disease prevention and health promotion efforts through improving early detection of children at risk for infection, optimizing timing of prevention efforts, and ultimately preventing adverse health outcomes from SSI. The methods employed in this study will also advance NICHD’s aspirational goals to leverage machine learning and artificial intelligence for precision medicine. The proposed training, guided by an expert mentorship team, will enrich the applicant’s skills in machine learning and prediction, translational data science for precision health, and clinical informatics and technology implementation. The applicant will benefit from interdisciplinary expertise, directed mentorship, and coursework from both the University of California, San Francisco and Stanford Medicine Children’s Health, two world-class research and clinical environments. This research and training will prepare the applicant for a future career as an independent researcher focused on optimizing pediatric health through evidence-based EHR tools with real-word impact on patient care and outcomes.

NIH Research Projects · FY 2025 · 2024-05

PROJECT SUMMARY Membrane tension is thought to be a long-range integrator of cell physiology. During migration, membrane tension has been proposed to enable cell polarity through front-back coordination and long-range protrusion competition. These roles necessitate effective tension transmission across the cell. However, it remains a source of significant debate as to whether cell membranes support or resist long-range membrane tension propagation. I speculated that this this discrepancy likely originates from the use of exogenous forces that may not accurately mimic endogenous forces. To overcome this complication, I used optogenetics to directly control localized actin-based protrusions or actomyosin contractions while simultaneously monitoring the propagation of membrane tension using dual-trap optical tweezers. My results led me to propose a unifying model of tension propagation in which actin-driven protrusions and actomyosin contractions both elicit rapid global membrane tension propagation, while forces applied to cell membranes alone do not. This work laid the foundations of my ongoing studies and raises many important questions. For some processes, membrane tension needs to act locally and in others, it needs to act at the range of the entire cell or multiple cells. What controls the range and efficiency of tension propagation (Aim 1)? The cell has many processes that can locally initiate changes in membrane tension (protrusions, contractions) and many cellular processes that are regulated by tension changes. Are there important functional differences for how these cellular programs transmit or receive membrane tension changes that may be important for cell polarity (Aim 2)? Migrating cells often need to coordinate their movement with other cells in order to achieve collective or cooperative motion. What is the impact of the membrane tension-polarity program in multicellular contexts such as encountered during collective or cooperative cell migration (Aim 3)? I will answer these questions by using a combination of optogenetics, force measurements, and advanced microscopy, and mathematical modeling. My research will elucidate the interplay between membrane mechanics and polarity during cell migration and will provide the foundation for my independent scientific niche of studying membrane mechanics during transendothelial migration. Towards this goal, I am supplementing the input of my postdoc supervisor, Dr Weiner (expert on cell polarity during migration) with a group of internationally recognized leaders at the interface of cell biology and biophysics: Carlos Bustamante (optical traps), Herve Turlier (mechanical modelling), Janis Burkhardt (cell migration). Japp van Buul and Ronen Alon, both experts on transendothelial migration. These scientists will act as both advisors and collaborators, helping me establish my own scientific niche in understanding the role of membrane mechanics in regulating transendothelial migration. I have worked with my mentor and network of collaborators and advisors to develop my research plan and construct a tailored career development program to help launch my independent career as a tenure-track principal investigator.

NIH Research Projects · FY 2026 · 2024-05

ABSTRACT: UCSF Understanding how T cell receptors (TCRs) see tumor antigens presented by MHCs is necessary to fully understand how the immune system recognizes tumor antigens, and to reap the full potential of antigen-specific immunotherapy. To achieve this goal, a quantum leap forward is required in which the revolutionary advances in machine learning are combined with a large volume of structure, function, data on matched TCR-pMHC pairs. The development of accurate predictors of TCR-antigen recognition will be dependent on the creation and integration sequencing-based datasets with high-throughput structural and functional insights. Our proposal, submitted as a CRUK/NCI Grand Challenge team (MATCHMAKERS) will combine researchers with expertise in immunology, methods development, structural biology, and computation to enable generalized prediction and design of TCR recognition. This work will be spread across four Work Packages (WPs): WP1: Large-scale generation of TCR-pMHC pairs from naturally occurring sources. We will build datasets of naturally occurring TCR-pMHC pairs. Our team will use an array of approaches to collect these datasets, from humans and from mouse models, and in the context of both cancer and immunity more generally. WP2: Ultra-high throughput TCR-pMHC matching using molecular engineering. Efforts to create general models will require a broader array of data than feasible to collect from natural TCR systems. We will use an array of synthetic approaches developed by our team to comprehensively match TCRs with pMHCs to train computational models. WP3: Large-scale structural and biochemical analyses of TCR-pMHC interactions. A key to our team’s vision is to match interaction datasets with high throughput structural and functional insights. A deep understanding of how the TCR contacts with MHC helices control function and orientation will be essential for training and testing computational models. WP4 AI-based prediction and design of TCR-pMHC interactions. We will integrate our data to train next- generation algorithms capable of generally predicting and designing TCR-pMHC interactions. These predictions will proceed through a reiterative testing and feedback circuits for further model optimization.

NIH Research Projects · FY 2026 · 2024-05

ABSTRACT Dry eye disease (DED) is a major health problem. Reduced ocular surface hydration is a key pathology in DED, which causes symptoms and can lead to ocular surface inflammation and damage. Current FDA- approved treatments for DED include artificial tears and anti-inflammatory agents that have variable and limited efficacy. Here we propose a novel target for treatment of DED. The target is the extracellular Ca2+-sensing receptor (CaSR) that has well-known modulatory effects on epithelial ion transport in various tissues such as intestine and kidney. We found prominent CaSR expression in mouse and human corneal and conjunctival epithelial cells. CaSR activation in the ocular surface reduced CFTR-mediated Cl- secretion, confirming key roles of CaSR ocular surface ion transport. Importantly, we found that CaSR inhibition by NPS-2143 stimulates CFTR-mediated Cl- secretion in the ocular surface and increases tear fluid volume in mice by ~100%. Here we will perform key translational studies for CaSR inhibitors, by testing them in clinically relevant animal and human cell models of DED to advance them towards clinical testing (Aim 1). We will also study the roles and mechanisms of CaSR effect in ocular surface ion transport and health (Aim 2). The proposed studies can potentially lead to clinical testing of CaSR inhibitors for DED in the very near future. By identifying a novel treatment, these studies can ultimately reduce DED morbidity. Lastly, by identifying roles and mechanisms of CaSR in ocular surface, this project can also identify novel disease mechanisms for DED.

NIH Research Projects · FY 2025 · 2024-05

Project Summary Combinatorial screens characterize the molecular pathways of the cell by identifying interactions between perturbations. Quantifying interactions requires a notion of non-interaction (i.e., a null interaction model), which is well-established for combinatorial screens measuring cell survival and other simple phenotypes. However, recent advances in single-cell omics and automated microscopy have enabled combinatorial screens with high- dimensional phenotypic profiles. These phenotypic profiles consist of features with unknown, non-linear behavior, such as gene expression or image features. As a result, there is no well-defined null interaction model for combinatorial screens with high-dimensional phenotypes. The goal of my proposal is to establish a null interaction model for high-dimensional phenotypes. My approach, termed DENIM (Data-driven Estimation of Null Interaction Models), aims to use robust estimation models to learn an additive representation for null perturbations and improve quantification of interactions across combinatorial screen datasets. In addition, this framework has a key application in drug target identification by enabling accurate prioritization of chemical-genetic interactions from microscopy-based phenotypic profiles. My central hypothesis is that null interaction models which accurately model the diverse behavior of high- dimensional features can substantially improve interaction detection. In Aim 1, I will develop and critically assess DENIM using both simulated and real-world combinatorial datasets with gene expression or high-content imaging readouts. I will evaluate DENIM against existing baselines on the ability to accurately interaction strength in simulation, as well as recapitulate known interactions between gene pairs or drug pairs. Preliminary data from simulation demonstrates that DENIM can accurately identify and learn non-linear perturbation trajectories, which leads to improvement in performance relative to the standard baseline (i.e., vector addition). In Aim 2, I will leverage the improved prioritization of interactions provided by DENIM to perform drug target identification. This target identification platform requires large-scale genetic perturbations and the study of chemical-genetic interactions. To this end, preliminary results show that large-scale genetic perturbations can be accurately conducted using pooled genetic screening followed by in situ sequencing for guide demultiplexing. Moreover, I analyzed a pilot screen on a well-established MEK1/2 inhibitor, where I accurately recovered target pathway components, with similar performance using drug-relevant or drug-agnostic biomarkers. Together, completion of this proposal will result in a rigorous framework for high-dimensional interactions, benchmarking of null interaction models, and a novel platform for chemical-genetic target identification using microscopy-based phenotypic profiles.

NIH Research Projects · FY 2025 · 2024-05

Project Abstract / Summary Emerin (EMD) gene mutations cause Emery Dreifuss Muscular Dystrophy type I (EDMD1), a disease characterized by skeletal muscle wasting and life-threatening atrial defects. However, the mechanism of emerin dysfunction in the heart is unknown. In this proposal, I seek to understand emerin's role in human atrial cells. Emerin is a protein of the inner nuclear membrane (INM), and a component of a nuclear cytoskeletal structure known as the nuclear lamina. Investigators have thus assumed that EDMD1 arises from defects in emerin's nuclear functions. However, Emerin has been observed at two locations in cardiomyocytes (CMs); emerin also localizes to the cardiomyocyte intercalated disc (ID)—a specialized plasma membrane (PM) domain that electromechanically couples cardiomyocytes to one another. Both of emerin's observed locations are essential for CM gene regulation and biomechanics, though we do not understand emerin's role at either structure. Our understanding has been limited by 1) the lack of tools to manipulate emerin's distribution, and 2) the lack of models to study these CM-specific mechanisms. This proposal will overcome these roadblocks by manipulating emerin's localization in human induced pluripotent stem cell-derived atrial cardiomyocytes (hiPSC-ACMs). Until our recent discovery, we did not know how emerin localizes to the PM, which impeded our ability to test its functions there. However, we have discovered that emerin can traverse the secretory pathway through vesicular transport to the plasma membrane. We hypothesize that secretory trafficking routes emerin to the ID to perform a specific function in cardiomyocytes. To test this hypothesis, I will first engineer a model cell system to assay emerin dynamics. Emd -/- mice do not display EDMD1 phenotypes and are thus a poor model of emerin pathology. hiPSC-ACMs, however, are tractable, mutable, and applicable to human disease. I have previously identified motifs within emerin that control its trafficking. Using this knowledge, I will introduce a panel of mutants with enhanced or blocked trafficking into emerin knockout hiPSCs. Using the cell lines I generate, I will test how emerin disruption affects ACM gene regulatory networks (Aim 2) and cellular mechanics (Aim 3). By controlling emerin's secretory trafficking, I will also test how its localization at the PM or INM influences these functions. This system will illuminate emerin's fundamental roles in human atrial cardiomyocytes for the first time. By defining these mechanisms, we will better understand EDMD1 cardiac defects and open the door to new potential therapies.

NIH Research Projects · FY 2024 · 2024-04

SUMMARY ABSTRACT Paralytic shellfish poisoning (PSP) is a serious illness that disproportionately affects Alaska Native communities. Sitka Tribe of Alaska has organized an environmental monitoring program and seafood safety testing program along with other regional partners (called SEATOR: Southeast Alaska Tribal Ocean Research) to reduce the burden of PSP disease in Southeast Alaska. This grant seeks to augment SEATOR's ongoing efforts, through additional data collection and partnerships with the Sitka School District and Emory University. Collecting additional years of PSP toxin monitoring data using the same protocols as today will allow a time- series analysis to inform a cutting-edge forecasting model that will give Southeast Alaska residents warning of especially risky subsistence shellfish harvest times, and thereby prevent poisonings. Collecting new data on specific kinds of PSP toxins (rather than only measuring the presence or absence of any PSP toxin) can clarify which other settings around the world are most similar (e.g., by comparing toxin profiles). This is important because PSP is a global health problem and there are other locations that could benefit from an informative prediction model, or that might be able to provide useful information for developing this region's model. SEATOR will continue training members of the community via workshops on how to collect and process samples per the citizen science framework of the ongoing work. This project also includes planned K12 educational interventions to target PSP prevention messages to youth at particular risk of PSP toxicity, while also promoting science education and traditional lifeways. This project is fully community-based participatory research (CBPR) building on existing tribal organization programs, with employment and skills-training opportunities for members of the community, to meet community-prioritized needs and to address this health disparity.

NIH Research Projects · FY 2026 · 2024-04

Project Summary/Abstract Type 2 diabetes (T2D) is a chronic degenerative cell disease characterized by hyperglycemia due to progressive progressive loss of overworked pancreatic islet beta cells. Recent work from our laboratories has shown that hyperactivation of the unfolded protein response (UPR) to ER stress in beta cells due to upstream may be a critical early event in the development of T2D. We have developed and are refining novel small molecule pharmacological reagents that allow us to manipulate critical components of the UPR—called KIRAs that maintain desirable (adaptive) outputs of the UPR while quelling undesirable (maladaptive) outputs including insulin resistance in peripheral tissues, consequently leading to apoptosis and diabetes. We show that first generation versions of KIRAs we developed against a kinase/RNase target called IRE1α can efficaciously prevent and even reverse diabetes in leptin deficient ob/ob mice. Here, we plan a medicinal chemistry campaign using several strategies to optimize KIRAs on three scaffolds towards the clinic. In this collaborative grant we will capitalize on the complementary expertise of the investigators to optimize these KIRA lead molecules for potency, selectivity, drug-likeness (including oral bioavailability) and conduct critical proof-of-concept studies using reductionist chemical genetic systems, ob/ob and HFD-fed mice, and human islets, and perform key IND-enabling steps needed to advance these candidates further into the clinic for eventually treating human patients with T2D.

NIH Research Projects · FY 2026 · 2024-04

Summary / Abstract The nuclear Lamins are intermediate filament proteins that underlie the nuclear envelope. Lamins assemble into bundled filaments that strengthen the nucleus and scaffold the genome. The LMNA locus encodes the Lamin A and C proteins; hundreds of mutations to LMNA have been linked to more than 10 distinct syndromes, collectively referred to as “laminopathies”. Disease-linked LMNA mutations are found in every domain of these proteins, and no clear genotype-phenotype correlations exist. LMNA-linked laminopathies primarily afflict the cardiovascular system, muscle, skin, connective tissue, bone, and fat. A major goal of our research program is to determine why specific tissues are especially vulnerable to LMNA mutations while the Lamin A/C proteins are broadly expressed. In this proposal, we present evidence that modulation of protein lifetime is a major novel axis that regulates both the function of Lamin A/C and the effects of laminopathy mutations. We have discovered that the lifetime of the Lamin A/C proteins varies over an order of magnitude across tissues, and these proteins are especially long- lived in the tissues that are most vulnerable to laminopathies. In the devastating laminopathy Hutchinson-Gilford progeria syndrome (HGPS), production of a toxic Lamin A isoform called “Progerin” disrupts nuclear function and causes accelerated aging. We determined that this mutation further slows the turnover of Lamin A in the diseased cardiovascular system, where it accumulates over time and interferes with the turnover of hundreds of other proteins. These findings raise key questions that will be addressed by our Aims. Firstly, while hundreds of mutations to LMNA are linked to human disease, we do not yet know how these other mutations affect Lamin A/C protein stability and/or aggregation within diseased tissues. We will leverage a novel in vivo metabolic labeling approach that we developed to probe the effect of another laminopathy mutation on protein turnover in the LmnaH222P mouse model (Aim 1). Secondly, we do not understand how Lamin A/C protein lifetime is modulated across tissues or in disease states. Differences in protein folding and/or assembly state often underlie differences in protein turnover rate, but we know very little about the structure of the Lamin A/C polymers in the cellular milieu or how variable these are across different tissue contexts. In Aim 2, we will use functional proteomics to probe the folded state of the Lamin A/C proteins in situ, and we will test the hypothesis that Lamins sense mechanical information – such as tissue strain – to alter their folded state. Thirdly, we know that the Lamins perform the important function of scaffolding heterochromatin at the nuclear periphery into repressive lamina-associated domains (LADs), yet we do not understand the consequences of variable Lamin A/C folded state on this Lamin function. In Aim 3, we will apply a novel genome binding analysis technique to map LADs and to define their repressive function within healthy and diseased tissues.

NIH Research Projects · FY 2026 · 2024-04

PROJECT ABSTRACT / SUMMARY There is a fundamental gap in our understanding of outcomes related to third-line treatments [onabotulinumtoxinA, peripheral tibial nerve stimulation (PTNS), and sacral neuromodulation] for refractory overactive bladder (OAB) in older and frail older adults, who collectively comprise the majority of patients suffering from this condition. Despite being well-studied in young and middle-aged healthy adults, little is known about outcomes and complications of these treatments in these more vulnerable populations, potentially resulting in overutilization of treatments that are ineffective or dangerous, or in underutilization of treatments that may significantly improve health related quality of life. Our overarching research objective is to improve care for all older and frail older adults undergoing treatment for OAB. The objective of this proposed study is to better understand the utilization, outcomes and complications associated with onabotulinumtoxinA, PTNS, and sacral neuromodulation in older and frail older adults, and to use this information to create an individualized and actionable prognostic tool. The central hypothesis is that utilization of these procedures may be biased towards younger aged individuals (of white race and higher socioeconomic status, performed by high volume specialists), regardless of level of frailty, but that frailty will be highly correlated with poor outcomes and increased rates of complications. We will test this hypothesis by leveraging a 100% cohort of fee-for-service Medicare beneficiaries undergoing these procedures and the claims based frailty index (CFI), as the measure of frailty, by the following three aims: (1) to understand the utilization of invasive OAB treatments (a) according to age and frailty, and (b) according to other non-clinical factors: patient (race/ethnicity), regional (socioeconomic) and provider (surgeon and volume); (2) to determine downstream outcomes and utilization of third-line OAB treatments across the spectrum of advancing age and frailty up to 2 years after the procedure; (3) to develop and internally validate a prognostic tool to predict successful outcomes for older and frail older adults undergoing third-line OAB treatments. This study is innovative because it will measure and apply the important factor of frailty to a national cohort of 100% of fee-for-service Medicare beneficiaries undergoing onabotulinumtoxinA, PTNS, and sacral neuromodulation to ultimately create an individualized prognostic tool to improve outcomes in this understudied population. The proposed research is significant because there is a critical lack of information about outcomes for refractory OAB in this large and vulnerable population who suffer greatly from this condition. Development of an individualized prognostic tool to aid in this decision-making process will serve to minimize the risks of potentially unsuccessful, unnecessary and even harmful procedures, while promoting the use of such procedures among individuals who are more likely to receive benefit.

NIH Research Projects · FY 2025 · 2024-04

Project summary/Abstract Our long-term objectives are to define the complex mechanisms responsible for brain damage and repair following neonatal hypoxia-ischemia (HI), as a model for neonatal hypoxic-ischemic encephalopathy (HIE) in term infants, and to search for novel and specific diagnostic and therapeutic targets for HIE. Inflammation is a hallmark of HI brain injury, during which brain resident microglia play critical roles in both induction and resolution of the inflammatory responses. We recently observed a striking buildup of cholesterol- containing lipid droplets in microglia (lipid-laden microglia) in the injured areas following HI in the neonatal mice, as well as in human HIE patients. In this proposal, we study how abnormal lipid accumulation and defective clearance of cholesterol in microglia affect their normal function and neuroinflammation after neonatal HI. We focus on cholesterol 25-hydroxylase (ch25h), which converts cholesterol to 25-hydroxycholesterol for clearance, and plays multiple roles in reducing intracellular cholesterol overload. We hypothesized that ch25h is critical for the maintenance of microglial homeostasis and attenuation of neuroinflammation following neonatal brain HI. In Aim1, we will assess how ch25h regulates microglial lipid accumulation and function in HI-injured neonatal brain. We will delete ch25h selectively in microglia and investigate the direct effects on lipid accumulation in microglia, microglial phagocytic ability, as well as the production of cytokines and reactive oxygen species. The contribution of microglial ch25h to brain injury are determined at early phase after HI. In Aim 2, to begin dissecting the mechanisms by which ch25h regulates microglia phenotype after neonatal HI, we will characterize the transcriptional profiles of microglia with unperturbed or deficient ch25h in the sham and HI-injured brain using translating ribosome affinity purification (TRAP) technology followed by RNA-sequencing. This is the first study of the crosstalk between cholesterol metabolism and microglial immunity using a clinically relevant mouse model of HIE. By addressing this important gap in knowledge in perinatal brain injury, we will improve our understanding of how lipid regulation/processing in microglia controls their innate immune and inflammatory responses, and contributes to brain damage/recovery. The results of this study could inform novel therapeutic opportunities for neonatal brain injury by targeting microglial lipid metabolism.

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY Inflammation promotes regeneration of epithelial tissues, but it is also implicated in intestinal pathologies including cancer, inflammatory bowel diseases and celiac disease. Recent studies in barrier tissues (i.e. skin and airway epithelium) demonstrate that stem cells in these organs maintain a memory of inflammatory exposure, which enhances their wound healing capacity in later injuries. Whether intestinal stem cells (ISCs) remember encounters with inflammatory stimuli, and how this affects their subsequent function, is poorly understood. Our group recently demonstrated that infection with the enteric helminth Heligmosomoides polygyrus (Hp) generates inflammatory granulomas in the murine intestine, which alter ISC identity and induce a fetal-like transcriptional signature in granuloma-associated crypts (GACs). Furthermore, we observed that ISCs isolated from GACs maintain this fetal-like identity when cultured as organoids ex vivo, indicating that they retain a memory of their exposure to helminth infection. The goal of this application is to investigate whether this inflammatory memory is encoded through epigenetic reprogramming of ISCs and how this process impacts their future regenerative capacity. In Aim 1, I will perturb candidate molecular regulators of inflammatory memory identified through analysis of epigenetic datasets comparing enteroids generated from non-inflamed tissue and fetal-like spheroids generated from GACs. Next, I will examine the in vivo epigenetic landscapes of intestinal crypts from naïve, Hp-infected, and previously infected/recovered mice using the 10x genomics Multiome assay. This will enable characterization of cell states across conditions and identification of open chromatin domains that are established during inflammation and retained after its resolution. Analysis of transcription factor motifs associated with these regions will provide additional insight into the signaling pathways that control this phenomenon. In Aim 2, I will study the downstream effects of these epigenetic changes on ISC function. I will first test whether ISCs that had prior exposure to inflammation have increased organoid forming capacity in vitro. I will then investigate whether previously inflamed tissue has faster regeneration kinetics in vivo, by inducing a second injury and assessing tissue morphology, proliferation, and differentiation. This study will improve our mechanistic understanding of ISC function in regeneration and provide insight into the role of inflammatory memory in intestinal homeostasis and pathology. The proposed research will be carried out under the guidance of a multi-disciplinary co-mentorship team with expertise in ISC biology and allergic inflammation within the dynamic and collaborative environment of UCSF. The proposed research plan is integrated with a comprehensive training plan which involves building conceptual knowledge in intestinal biology and immunology, and developing technical proficiency in the use of transgenic animal models, epigenomic assays, and bioinformatic analyses. During this fellowship, I seek to advance my mentorship, networking, and career skills to ultimately enable a successful transition to an independent investigator position at a top-tier academic institution.

NIH Research Projects · FY 2026 · 2024-04

This D43 grant application aims to sustainably strengthen the research capacity of the Universidad Javeriana in Bogotá, Colombia with an emphasis on training in-country experts to develop and conduct research focused on Alzheimer’s disease and related dementias (ADRD). Our application is in response to NOT-AG-21-027 from the National Institute of Aging (NIA) aimed at encouraging the development of training and research programs related to ADRD in low- and middle- income countries (LMICs). We build on existing research collaborations, funded by the National Institute of Aging, in the Multi-Partner Consortium to Expand Dementia Research in Latin America (ReDLat, R01-AG-057234) between UCSF and Universidad Javeriana. We also expand UCSF’s Global Brain Health Institute’s (GBHI) eight-year engagement in Colombia by training five Atlantic Fellows in a non-residential fashion to advance ADRD research, education, and care. Three pillars of activities described in this application will be used to increase ADRD research capacity for the university framed around a new Doctorate of Neurosciences program led by MPI, Santamaría-García. First, we will augment programing for a new Doctorate of Neuroscience degree with further coursework in statistics and grant writing. A full-time statistician at Universidad Javeriana will amplify the success of this program. We will support the most promising doctorate students with stipends and seed grants to free up time needed to advance their research. Second, we will increase capacity of the early-career mentoring pool through a non-residential experience provided by GBHI and will provide access to seed grant funding for early-career, patient-oriented faculty at Universidad Javeriana. Most seed grants to faculty will be offered in year 1 to create a platform for research projects that postdoctorate students can join and to provide preliminary data for NIH applications, a core metric of success. The non-residential experience at GBHI will be 12 months in duration and will include coursework around ADRD, leadership, mentoring, and skills such as grant writing. Resources for these alumni include access to USD 25,000 pilot grants program. Finally, we will increase in-country networking and research exchange opportunities, in collaboration with the Universidad de Antioquia which has a robust NIH-supported research program around early-onset, dominant-inherited AD. An in-country annual scientific exchange and workshop will fortify these collaborations. In all, the proposed work aims to substantially fortify Universidad Javeriana’s research capacity for ADRD research in a manner that strengthens the renewal of this grant in five years led by MPIs in Colombia.

NIH Research Projects · FY 2026 · 2024-04

Project Summary/Abstract The goal of this proposal is to elucidate biological mechanisms for proton transfer by designing function from scratch. The coupled movement of protons and electrons is crucial to biological energy transduction and central to life. While electron transfer (ET) has been extensively studied, less is known about the corresponding proton transfer (PT) due to lack of easily observable experimental readouts. Computational protein design enables us to study these phenomena in a ground-up manner where a protein scaffold can be designed from first principles to mimic biological function in isolated and experimentally tractable ways. This proposal centers on the binding of abiological photoacid cofactors that would give distinct spectroscopic readouts for PT as a function of distance. The electron-deficient metal porphyrin photoacid cofactors used in this proposal are characterized by dramatic acidification upon photoexcitation and distinct spectroscopic changes upon deprotonation. These cofactor properties combined with our lab’s history of success in the design of porphyrin-binding proteins make them ideal for use in this proposal. Using computational tools recently developed in the DeGrado lab (vdMs and COMBS), the cofactor will be positioned within a designer protein scaffold H-bonded to a proton-accepting residue. This will enable the spectroscopic study of proton on-off rates upon irradiation and subsequent deprotonation of the cofactor. These ligand-binding proteins will be experimentally characterized through X-ray crystallography and NMR experiments to validate the proposed structure and binding mode. Ultrafast absorbance spectroscopy experiments will be carried out by our long-term collaborators in the Therien lab at Duke University. Following characterization, the proton-accepting residue will be iteratively moved down the protein scaffold with a designed “water-wire” in its wake to allow spectroscopic observation of the proton movement over varying distances. Further, a second cofactor binding site will be built to bind a pH-responsive dye. This will allow for end-to-end monitoring of PT with measurable readouts in a protein system for the first time. This research will significantly advance our understanding of biological proton dynamics, critically test our ability to design ligand-binding proteins, push toward the intentional design of water wires, and innovate a new strategy for the design of proteins that bind multiple interacting cofactors. The use of computational design tools (Rosetta, COMBS, RFdiffusion, ProteinMPNN, Alphafold), as well as routine protein expression, purification, and characterization will fulfill the training goals of my postdoctoral tenure, combining my skills in organic synthesis with protein design. Together these skills will prepare me for an independent research career focused on the design of functional proteins and enzymes to catalyze new-to-nature reactions.

NIH Research Projects · FY 2025 · 2024-04

Project Abstract Biomedical research scientists face unique career challenges that, if unaddressed, hinder the success, well-being, and ultimately retention of this specialized workforce. Curricular support to meet these threats to well-being first requires a comprehensive understanding of the diverse experiences of biomedical research scientists, across specialties and career stages. After conducting interviews with biomedical research program directors, and focus groups with biomedical faculty and research trainees across our institution, we will develop a set of modules to address well-being needs along four tracks: a Foundational track that provides content on the Six Areas of Worklife and how they bear on well-being; a Resilience track that provides evidence-based tools for formulating a personal plan for well-being and navigating the rejection that is inherent to a successful biomedical research career; a Relational Support track that identifies strategies for developing healthy mentorship and sponsorship relationships and techniques for collaborative research success, and a Programmatic Infrastructure track that includes support for navigating differences, identity formation, and building capacity for adaptability. A series of facilitator guides will accompany the modules to allow for consistency in interactive content. Each module will be evaluated for its educational value through mixed- methods investigation, and in consultation with an advisory board composed of graduate education experts, directors of educational assessment, leadership from the UCSF Office of Career and Professional Development, diversity and learner success, student and faculty academic affairs, faculty equity, and a Chief Wellness Officer. A randomized controlled study design will allow for assessment of the impact of training materials on validated well-being, resilience, career self-efficacy and adaptability measures. The training materials will be made freely available and will provide a sustainable set of tools for building well-being, resilience, and adaptability in our biomedical research scientist workforce.

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY The sinoatrial node (SAN) is a tiny structure consisting of a precise arrangement of specialized pacemaker cardiomyocytes (PCs) that trigger each heartbeat. Sinus node dysfunction (SND), resulting from loss or malfunction of PCs, is a common and morbid disease that is not well understood. There is currently no treatment that can delay or prevent SND, so sufferers must undergo permanent pacemaker implantation. This proposal will leverage novel tools and experimental approaches to determine how a multipotent population of SAN progenitor cells are allocated to different fates within the SAN, and to define how transcriptional hierarchies govern gene expression programs and spatial organization in the SAN. Ultimately, we hope that our findings will lead to new approaches to prevent or reverse SND by targeting the biological pathways that control SAN formation and maintenance. Previous efforts in this area have been hampered by the lack of specific genetic tools to mark the SAN progenitor population and the lack of an in vitro system that can model SAN development. In the past, our group has co- discovered a key activating role for the transcription factor Isl1 in SAN development and we have identified an Isl1 SAN Enhancer (ISE) that is specifically active in the SAN and its progenitor population. We have used this enhancer to generate a new mouse line, ISE-CreERT2, that allows conditional genetic modification of the SAN progenitor population. Clonal fate mapping with this mouse line and single cell sequencing analysis have defined the dynamics of the SAN progenitor population and allowed us to develop a system to explore how SAN progenitors are allocated among several possible fates. To gain further insight into how this fate allocation occurs, we developed a novel in vitro protocol using hiPSCs that recapitulates key aspects of SAN differentiation, including diversification of SAN progenitors into anatomically and functionally distinct pacemaker cardiomyocyte subtypes. Using this model, with in vivo studies as validation, the present work explores the hypothesis that SAN progenitor allocation depends upon Isl1, activator protein-1 (AP-1), and Nuclear Factor I (NFI) transcriptional programs that guide cells in the SAN progenitor field to different fates. In Aim 1, we will determine how Isl1 shifts from playing a role in regulating cardiac progenitor cell proliferation to playing a key role in activating the PC-specific gene expression in a cellular subtype-specific manner. In Aim 2 we will determine how AP-1 and NFI regulate progenitor cell allocation and adoption of cell type-specific gene programs in the SAN. Aim 3 will determine how these pathways regulate the spatial architecture of the SAN using spatial transcriptomics in WT and genetically modified mice. Taken together, the work proposed will establish new mechanistic paradigms for the how the mammalian cardiac pacemaker is formed and could set the stage for novel approaches to treating and preventing SND.

NIH Research Projects · FY 2026 · 2024-04

Abstract Antibody-drug conjugates (ADCs) have shown promise as targeted therapeutics for treating cancer, including solid tumors. However, no ADC has gained FDA approval for treatment of prostate cancer. Multiple factors including target selection and heterogeneity, as well as insufficient tumor uptake and intracellular delivery, could have contributed to the limited success of current ADCs. To overcome these limitations and improve treatment outcomes, we propose to develop novel bispecific ADCs (bisADCs) that target emerging tumor cell surface antigens (CD46, B7-H3, and DLL3) with clinical validations. Bispecific targeting, which engages two distinct tumor antigens simultaneously, holds promise for overcoming target heterogeneity, improving internalization and tumor penetration. Unlike a combination of monoclonal ADCs, bisADCs possess the unique ability to exploit synergistic interactions between targets and influence their intracellular fate. This concept provides a compelling rationale for exploring bisADCs in prostate cancer treatment. We discovered CD46 as a novel prostate cancer cell surface antigen that is persistently expressed across differentiation patterns. We developed and translated a CD46-targeted ADC (FOR46) to phases I (NCT03575819) & II (NCT05011188) trials. This CD46 ADC showed good tolerability and promising early efficacy signals in mCRPC patients. B7- H3, another emerging target widely expressed in prostate cancer, has been targeted with ADCs like MGC018 and DS-7300, showing acceptable safety and encouraging efficacy in early-phase trials. B7-H3 negativity is rare in adenocarcinomas but more common in CRPC with neuroendocrine features. DLL3 is overexpressed in several neuroendocrine tumors, and a DLL3-targeted ADC (Rova-T) has been tested in neuroendocrine prostate cancer in a phase I trial, making DLL3 a credentialed target for this subtype. We propose to develop bisADCs based on those emerging novel targets to address both adenocarcinoma (B7-H3 x CD46) and small cell neuroendocrine (DLL3 x CD46) prostate cancer. In Aim 1, we will determine target co-expression patterns in mCRPC patient samples, generate novel human monoclonal antibodies and nanobodies against the targets of interest, and construct bispecific antibodies (bisAbs) using robust Ig-like architectures, and generate bisADCs carrying diverse drug payloads with clinical validation. In Aim 2, we will investigate the therapeutic efficacy of B7-H3 x CD46 and DLL3 x CD46 bisADCs in animal models representing adenocarcinoma and small cell neuroendocrine prostate cancer, respectively. We will benchmark the bisADCs against current ADCs being tested in clinical trials, and identify lead bisADCs that demonstrate improved efficacy against heterogeneous tumors, including target-low tumors, and enhanced survival in animal models of prostate cancer. The proposed study presents a novel approach to next-generation ADC development and has a clear path of translation to the clinic in prostate cancer patients who have progressed beyond current therapies.

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY/ABSTRACT Children with advanced neuroblastoma (NBL) have poor outcomes despite aggressive, multimodal therapy and new therapeutic approaches are needed. Recent studies have demonstrated that targeted 131I-MIBG radiopharmaceutical therapy combining modulators of somatic epigenetic programs (vorinostat) and GD2- targeted immunotherapy (dinutuximab) show substantial therapeutic promise. Although 131I-MIBG is one of the most effective therapies for advanced NBL, not all patients will respond to this therapy and predictive biomarkers of treatment response and mechanisms of resistance have not been identified. Our preliminary work in NBL show that minimally invasive liquid biopsy strategies for the detection of circulating tumor DNA (ctDNA), circulating tumor cells (CTCs), and cell-free GD2 levels can be successfully employed in longitudinal samples without reliance on surgical procedures in this vulnerable pediatric population. In this proposal, we will apply these technologies to samples collected from a randomized, phase 2 trial designed to identify the optimal agents for combination with 131I-MIBG in relapsed/refractory NBL (NANT 2021-02 [N21-02]) launching soon. Patients are randomized to either (A) vorinostat + 131I-MIBG; (B) dinutuximab + 131I-MIBG; or (C) vorinostat + dinutuximab + 131I-MIBG. Our overall hypothesis is that circulating biomarker levels will be associated with treatment response for each combination being tested in this trial. We further hypothesize that selection for mutations, changes in chromatin signatures, and downregulation of antigen expression will be associated with disease progression. In Aim 1, we will profile serial plasma samples to detect and quantify ctDNA levels and measure the association between ctDNA levels and response to therapy on each arm of N21-02. Furthermore, we will identify mutations associated with the development of treatment resistance through deep sequencing of ctDNA samples. In Aim 2, we will validate our existing chromatin accessibility and gene expression signatures associated with vorinostat treatment in preclinical models and in vorinostat-treated patient samples. We will then leverage serial liquid biopsy samples from patients enrolled on N21-02 treated with and without vorinostat to detect induction of vorinostat signatures to determine whether these signatures are associated with responses to vorinostat-containing therapies. Finally, in Aim 3, we will longitudinally measure soluble GD2 levels and surface expression levels on CTCs in samples collected from patients on dinutuximab-containing treatment arms and compare GD2 levels to patients not receiving dinutuximab therapy. We will test the association of soluble GD2 levels and response to GD2-directed therapy and determine whether changes in GD2 expression on CTCs are associated with the development of disease progression on therapy. At the end of this award, we expect to identify biomarkers of response and mechanisms of treatment resistance in patients with relapsed NBL. These findings will lead to optimized treatment selection and new candidate therapies to prevent the emergence of resistant disease in patients with NBL.

NIH Research Projects · FY 2026 · 2024-04

Cerebral palsy (CP) encompasses a group of disorders of movement and posture, attributed to non-progressive disturbances affecting the developing brain. Movement and tone disorders in children and young adults with CP are a great source of disability. CP is the most common cause of acquired dystonia in childhood, but its management is problematic, as medications and neurotoxin denervation only provide modest benefit. Deep brain stimulation of basal ganglia or thalamic targets has a major role in the treatment of isolated dystonias, but its efficacy in dystonic CP (DCP) is much lower. Lower efficacy may be due to structural damage in basal ganglia and motor thalamus, lack of improvement of comorbid choreoathetosis and spasticity, and an increased risk of hardware complications in this population. We propose an alternative brain target for DBS in DCP, the cerebellum, leveraging recent developments in dystonia neurophysiology, brain stimulation hardware and neuroimaging. The cerebellum is an attractive target for DBS in DCP since it is frequently spared from hypoxic ischemic damage, it has a prominent role in contemporary network models of dystonia, and small studies have shown promise of cerebellar stimulation in improving spasticity and CP-related movement disorders. Ten children and young adults with CP and disabling movement disorders and spasticity will undergo bilateral DBS in dorsal (motor) dentate nucleus, with the most distal contact in superior cerebellar peduncle. We will implant Medtronic PerceptTM, an FDA-approved “bidirectional” neurostimulator that can sense and store brain activity as well as simultaneously deliver DBS therapy. Recent improvements in hardware size and brain lead fixation address the prior high complication rate reported for pediatric DBS, and provide “directional” leads for more targeted stimulation. We will characterize abnormal patterns of cerebellar oscillatory activity as measured by local field potentials (LFP) related to clinical assessments and wearable monitors, and their relation to stimulation (Aim 1). Pre- and 12-month postoperative volumetric structural and functional MRI and diffusion tensor imaging will be used to identify candidate imaging markers of baseline disease severity and response to DBS (Aim 2). In Aim 3, we will test the efficacy of cerebellar stimulation for improving quality of life as well as motor outcomes as assessed by clinical assessments and objective kinematic metrics. We will use a N-of-1 clinical trial design to mitigate the variability in clinical features in this population. Our goal is to develop a neuromodulation therapy that produces meaningful changes in function and well-being of people with CP, obtain a mechanistic understanding of the underlying brain network disorder, and identify physiological and imaging-based predictors of outcome useful in planning further studies.

NIH Research Projects · FY 2026 · 2024-04

Title: CRISPRa-based rescue of sensorimotor deficits in the Scn2a+/- mouse model of autism spectrum disorder Project Summary: Heterozygous loss-of-function mutations in the sodium channel gene SCN2A are strongly associated with autism spectrum disorder. SCN2A encodes the neuronal sodium channel NaV1.2, which contributes to membrane excitability in somatodendritic or axonal compartments, depending on cell class. Heterozygous loss of Scn2a, modeled in Scn2a+/- mice, causes deficits in cellular excitability and synaptic function across multiple brain areas, and drives robust systems- and behavioral-level deficits in sensory processing, neural coding, and sensorimotor learning. Here, our goal is to rescue these cellular, circuit and behavioral deficits using CRISPR-activation (CRISPRa)-based therapeutic approaches in Scn2a+/- mice. Most autism risk genes, like SCN2A, are associated with autism by heterozygous loss-of- function. CRISPRa is a promising therapeutic approach for many genetic forms of autism because it upregulates expression of the remaining functional gene copy, potentially rescuing cellular function. Scn2a+/- mice are an ideal test case for CRISPRa intervention in severe genetic forms of autism. We have developed CRISPRa-based reagents that restore Scn2a expression to near- normal levels, and preliminary data show that treatment of adolescent Scn2a+/- mice with CRISPRa successfully rescues cellular and synaptic phenotypes in at least one brain area. Here, we will test the effectiveness of CRISPRa rescue of cellular, circuit, and behavioral phenotypes across sensory, associative, and motor regions of the brain. To do so, we bring together three laboratories with proven expertise in Scn2a disorders and autism, cellular and systems physiology, and therapeutics. Results of this study will help establish the feasibility of gene therapy for neurodevelopmental disorders including severe genetic forms of autism, and will help define the critical developmental windows in which therapeutic interventions are most effective.

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY Increased glucose and fat metabolism are essential for tumor progression. Recent work showed that cold- induced activation of brown adipose tissue (BAT), a unique adipose tissue which produces heat via glycolysis and fat metabolism, suppressed tumor progression in several cancer mouse models due to competition for glucose and fat resources. White adipose tissue (WAT) is an endocrine tissue that functions as the main energy storage organ in our body, storing lipids and secreting hormones that regulate various biological functions, including appetite, glucose and fat metabolism, and insulin hemostasis. It is commonly used in clinical procedures such as liposuction and fat transplantation in plastic surgery. Thus, WAT could be readily used for cellular therapy. Here, we propose to showcase the utility of this approach, which we term as Adipose Manipulation Transplantation (AMT), for cancer therapy. We plan to bioengineer WAT and adipose organoids to become more BAT-like, which will increase their glucose and fat utilization, along with having increased glucose uptake and fat storage. This will be done using CRISPR activation (CRISPRa) to upregulated genes that are involved in BAT function and glucose and fat metabolism. Our preliminary results already show that co- culturing CRISPRa engineered BAT-like adipocytes with five different cancer cell lines suppresses their growth. Furthermore, implanting engineered adipose organoids with xenografts or in two cancer genetic mouse models (breast and pancreas) significantly reduces cancer growth. Finally, engineering adipocytes from eight different human breast cancer patients surgically resected tissue and co-culturing them with cancer organoids significantly reduced organoid growth. Here, we plan to build on these results. We will engineer adipocytes and organoids to have increased glucose and fat utilization by upregulating combinations of genes not only involved in browning, but also in glucose and fat metabolism and glucose transport and test their ability to reduce cancer growth in various cancer cell models (Aim 1). We will also test the ability of these CRISPRa gene combinations in adipose organoids to suppress cancer in xenograft cancer models from various cell lines (breast, colon, pancreas, prostate) and in several genetic mouse models (breast and pancreas) (Aim 2). Finally, to further dissect the translational potential of this approach, we will use surgically resected tumor tissue from breast cancer patients, isolate and engineer their adipocytes and then co-culture them with their respective tumor to assess their therapeutic potential (Aim 3). This will be done across the full spectrum of human breast cancer stages, including different stages of breast cancer (adjuvant, late recurrence, metastasis, challenging-to-treat breast cancer subtypes and BRCA mutations). Combined, this project will develop a novel ‘CAR T like’ therapeutic approach to treat cancer utilizing bioengineered adipocytes, having tremendous therapeutic implications for cancer treatment.

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY: Distal radius fractures are one of the most commonly occurring fractures and represent a great morbidity burden in the United States and worldwide. Patient reported outcome measures (PROMs) that assess patients’ physical function are the standard for assessing treatment outcomes and can help inform treatment decisions for patients with distal radius fractures. Notably, however, PROMs must be linguistically and culturally relevant to appropriately measure physical function and inform care. There are currently no appropriately culturally adapted PROMs to assess the physical function of Spanish-speaking patients with distal radius fractures. As PROM scores affect care decisions, mismeasurement from the lack of cultural adaptation can affect clinical care and patient outcomes. The development and validation of culturally adapted PROMs for Spanish- speaking patients with distal radius fractures will yield new knowledge about gaps in the cultural adaptation of linguistically translated PROMs and provide the foundation for future R01 proposals to study the impact of cross-culturally adapted PROMs on measuring physical function, subsequent treatment decisions, and outcomes for patients with distal radius fractures and other orthopaedic injuries. By including patients from two countries, we can assess the impact of culture on outcome measurement which may be generalizable to patients in the United States and worldwide. This K23 Award will provide Dr. Shapiro focused training and mentorship in three critical areas: 1) conducting psychometric evaluations of PROMs using mixed-methods research designs, 2) understanding the cultural and linguistic influence on health and outcome measurement, and 3) conducting trials of outcomes measurement tools in diverse populations. Dr. Shapiro has assembled a team of mentors that includes: Dr. Patricia Katz (Professor Dept. of Medicine, UCSF), Dr. Alicia Fernandez (Professor Dept. of Medicine and Associate Dean, UCSF), Dr. Theodore Miclau (Professor Dept. of Orthopaedic Surgery and Vice Chair, UCSF). This is supplemented by a team of expert advisors and domestic and international collaborators. We aim to 1) assess the content validity of the Patient-Reported Outcomes Measurement Information System – Physical Function (PROMIS-PF) for measuring physical function in Spanish-speaking patients, 2) adapt the PROMIS-PF for two Spanish-speaking populations with distal radius fractures, and 3) validate and pilot test the new culturally-adapted PROMIS-PF tools. This will be the first study to evaluate the impact of culture on PROM items and to subsequently cross-culturally adapt, validate, and pilot test cross-culturally adapted PROMs using mixed-methods design for two Spanish-speaking populations. The new PROMs can serve as the foundation for future R01 proposals to study the impact of cross-culturally adapted PROMs on treatment decisions and outcomes.

NIH Research Projects · FY 2025 · 2024-04

PROJECT SUMMARY Diffuse midline gliomas (DMGs) are lethal brain tumors in children. Patient prognosis is very bleak with median overall survival of ~9 months. These tumors typically arise in delicate anatomical locations such as the brain stem that prohibit surgical resection. Radiation is the only standard of care and provides symptomatic relief. However, disease control is transient, and children inevitably face tumor recurrence and premature death. There is a need for novel therapies for DMG patients, especially those that can be combined with radiation. Glutathione (GSH) is essential for scavenging reactive oxygen species generated by radiation. GSH is also essential for the detoxification of methylglyoxal (MGO), a highly reactive metabolite that is spontaneously produced during glycolysis. MGO induces apoptosis by irreversibly damaging proteins and DNA, and cancer cells adapt to MGO production by upregulating expression of glyoxalase 1 (GLO1), which detoxifies MGO by conjugation with GSH. Analysis of TCGA data as well as our preliminary studies with patient-derived DMG models indicate that GLO1 expression is upregulated in DMGs. Inhibiting GLO1 using the potent brain-penetrant GLO1 inhibitor S-p-bromobenzylglutathione cyclopentyl diester (BBG) abrogates MGO detoxification and inhibits proliferation of patient-derived DMG cells. Importantly, while BBG alone arrests tumor growth in mice bearing orthotopic patient-derived DMGs, combined treatment with BBG and radiation induces tumor regression in vivo. Based on these results, in Aim 1, we will determine whether the combination of BBG and radiation is an actionable therapeutic strategy in patient-derived DMG models. Successful translation of novel therapies is aided by the identification of imaging biomarkers that report on early response to therapy. Deuterium magnetic resonance spectroscopy is a novel, clinically translatable method of visualizing the metabolism of 2H-labeled substrates in vivo. Our preliminary studies indicate that BBG downregulates lactate production from [6,6-2H]-glucose in DMGs. Therefore, in Aim 2, we will determine whether [6,6-2H]-glucose provides a readout of response to combined BBG and radiation, prior to MRI-detectable anatomical alterations, in mice bearing orthotopic DMGs in vivo. Our proposal is innovative because we will validate GLO1 as a druggable vulnerability in DMGs. This project is significant because our studies will set the stage for translation of the combination of BBG and radiation to DMG patients. Concomitantly, [6,6-2H]-glucose will enable early assessment of efficacy in clinical trials and in patient management. In essence, by leveraging metabolism for therapy and for imaging treatment response, we will deliver precision medicine that enhances outcomes and quality of life for DMG patients.

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY/ABSTRACT Autoimmune Regulator gene (AIRE) prevents autoimmunity by promoting thymic deletion of self-reactive T cells. While most studied in the thymus, AIRE is also expressed in secondary lymphoid organs, where it is thought to contribute to peripheral tolerance through interaction with and deletion of self-reactive T cells. However, AIRE’s role in extrathymic Aire-expressing cells (eTACs) is likely more multi-faceted than in the thymus. We recently identified AIRE expression in multiple myeloid populations in the tumor microenvironment (TME). These include dendritic cells (DCs) and monocytes, but are predominantly composed of tumor associated macrophages (TAMs), which are known to inhibit anti-tumor immune responses. While tumor associated eTACs were only recently discovered by our group, prior work implicates AIRE in inhibiting anti-tumor immunity. Recently available tools to study AIRE in mice have facilitated investigation of peripheral AIRE expression in the TME. We have found tumor associated eTACs in several common solid tumor models expressing myeloid lineage markers by both flow cytometry and mass cytometry (CyTOF). Critically, we also demonstrated that ablation of Aire+ cells dramatically slows tumor progression. This preliminary data together with the therapeutic potential for targeting eTACs in the TME to improve cancer immunotherapy make this a population deserving of thorough functional investigation. This proposal will test the hypothesis that tumor associated eTACs are an immunosuppressive, pro-tumoral cell population. Aim 1 of this proposal will define the phenotypic and transcriptional profiles of tumor associated eTACs, as well as their spatial organization. Aim 2 will determine the mechanism by which Aire+ cells can promote tumor progression. Aim 3 will define the signals that drive Aire expression and elucidate the cell-intrinsic role of Aire in tumor associated eTACs. This research approach will be carried out using a variety of methods including single cell analysis via RNA-seq, CyTOF, flow cytometry and in vivo assays utilizing novel genetic mouse models. These proposed studies will be the first characterization of AIRE expression in any tumor resident immune cells and will further our understanding of the function of peripheral AIRE-expressing cell types. This could result in the discovery of novel pathways relevant to therapeutic resistance and improve our understanding of global AIRE function. Translationally, this work may identify novel immune targets for cancer therapies. This research project and fellowship training will be conducted at a top-funded research institution, the University of California, San Francisco (UCSF), in the laboratories of Dr. James Gardner and Dr. Matthew Spitzer, with expert mentorship from Dr. Vasilis Ntranos. Dr. Gardner has expertise in the study of peripheral AIRE and mouse model generation. Dr. Spitzer has expertise in systems immunology and tumor infiltrating myeloid cell biology. Dr. Ntranos has expertise in computational analysis of single cell datasets. Overall, this facility and team provide a rich training environment for completion of this research and development of professional skills necessary for a career in academic research.

NIH Research Projects · FY 2025 · 2024-04

PROJECT SUMMARY A chronic inflammatory state, termed inflammageing, is a major driver of morbidity and premature death among older adults. Although the identification of inflammatory triggers has been identified as a critical area for aging research, little attention has focused on the skin, despite its role as a primary immunoregulatory organ. Normal aging is associated with increased skin barrier permeability, which leads to subclinical inflammatory cascades in the skin and serum. We hypothesize that age-associated decline of skin barrier function contributes to inflammageing and that restoring the skin barrier with moisturizers will reduce systemic inflammation. We propose a pilot trial designed to determine the best measures of skin barrier function in older adults, the most sensitive measures of systemic inflammation, the role of the cutaneous microbiome, and the feasibility and acceptability of moisturizer use. In a parallel, 8-week, self-controlled design, 32 subjects ≥70 years of age will be randomized to treatment with a ceramide moisturizing cream or petrolatum ointment. Changes in inflammatory markers, skin barrier function, and microbial diversity will be compared between a 4-week treatment period of moisturizer application and a 4-week placebo period. We will also examine recruitment efficacy, retention rates, fidelity to the treatment, data integrity, and the acceptability of the intervention and protocol. The results will be used to design a future randomized efficacy trial of moisturizers to reduce systemic inflammation. While many other pharmacologic strategies for reducing inflammation in older adults are considered too risky or expensive for widespread use, moisturizers are a promising intervention that is safe and accessible in diverse community settings.