University Of California, San Francisco

NIH Research Projects · FY 2025 · 2025-08

The overall goal of the UCSF SPORE is to improve cancer health outcomes by investigating the interplay of genomic and immune characteristics with individual risk factors and external drivers of health. Three cancers (meningioma, prostate cancer and breast cancer) with well-known differences related to clinical outcomes, will be studied. At the heart of the proposal are three translational research projects—each intended to evaluate novel tools and models to examine the interplay of external drivers and the biology of the disease with defined human endpoints. Project 1 aims to understand meningioma outcome differences between Black and White individuals in the context of external factors and individual-level risk factors using human samples, bioinformatics, preclinical models, and epidemiological data. Project 2 will characterize, at an unprecedented detail and depth, the explanatory variables, both biological (genomic) and environmental and individual lifestyle parameters, that explain prostate cancer outcome differences, with a particular focus on the complex interactions among stress and inflammation within across different populations. The response of the immune system to breast cancer therapy is a major determinant of breast cancer outcomes and the tumor immune microenvironment (TME) varies substantially among different populations. The goal of Project 3 is to identify the genetic and environmental factors that underlie these differences and to understand the underlying pathway mechanisms. Each Project is led by a balanced representation across basic and applied/clinical research investigators with critical involvement from biostatistical experts working with epidemiological scientists to ensure scientific rigor of the research strategy. Cancer surgeons are also integral to the projects. The Developmental Research and Career Enhancement Programs are vital to the expansion of research opportunities that will foster and build on a strong community of qualified investigators across the HDFCCC. The projects and programs will be supported by 3 specialized Cores (Administrative, Biospecimen and Pathology, and Community Outreach and Engagement). Research progress and scientific direction will be evaluated by the Executive Committee of the SPORE (all leaders and directors) with the expert guidance of the Internal and External Advisory Boards. In addition to leveraging the multidisciplinary expertise within the research programs in the UCSF HDFCCC, the 3 established UCSF SPORE grants in Brain, Prostate Cancer and Developmental Hyper-Ras Tumors (DHART) and the incredible resources from the institution, this SPORE will benefit greatly from rich collaborations with investigators at outside institutions and the Community Advisory Board members. The work proposed under this grant will yield valuable insights and make meaningful progress in understanding and reducing health outcome differences related to patients with meningioma, prostate cancer and breast cancer respectively.

NIH Research Projects · FY 2025 · 2025-08

ABSTRACT Down syndrome (DS) is the most common chromosomal disorder of intellectual disability, affecting approximately 400,000 people in the United States and approximately 5.5 million people worldwide. DS results from the presence of a third copy of chromosome 21 (Chr21) and associated genetic dosage changes that cause varying and complex disabilities throughout development and adulthood. With recent increases in the lifespan of persons with DS, it is now realized that Alzheimer’s disease (AD) is a substantial comorbidity in individuals with DS that arises at a much earlier age. Nearly all adults with DS develop severe AD neuropathological changes by age 40; however, there is evidence that the first appearance of amyloid-β plaques and tau neurofibrillary tangles (NFTs) occurs as early as the teens or twenties. Cryo-electron microscopy (cryo-EM) structural studies have revealed that the tau protein adopts a myriad of polymorphic folds that are associated with distinct etiological forms of tauopathy diseases across the spectrum of AD and related disorders (ADRDs). Recently, our novel workflow of in situ conformational assays and cryo-EM methods applied in aged patient samples revealed that DS tau adopts the AD folds of paired helical filaments and straight filaments. In our study, we developed a novel method using custom-made graphene-oxide (GO) grids functionalized with the anti-phospho tau “AT8” antibody to isolate and enrich filaments directly on cryo-EM sample grids. Using GO-AT8 grids, we isolated tau fibrils from a 36-year- old DS case with no dementia and few tau NFTs, thus demonstrating the potential for characterizing the structure of tau filaments at the onset of AD. Our proposal builds on our preliminary data and evidence in the literature suggesting there may be greater heterogeneity of tau filament structures at the inception of tau deposition and with certain neurological comorbidities, such as seizures and self-injurious, recurring head trauma. We hypothesize such comorbid conditions may lead to patterns of tau histopathology associated with chronic traumatic encephalopathy (CTE) and tau filaments with the CTE fold along with AD morphologies in a subset of individuals with DS. To facilitate the rapid success of this work, we will use a cohort of DS samples already in our labs via ongoing collaboration to screen a large number of fixed samples and prioritize a small number of frozen samples for cryo-EM. Further, we will expand our novel immunoaffinity approaches using tau conformation-specific antibodies to isolate putatively distinct tau conformers prior to cryo-EM. Therefore, we will implement the fluorescence imaging and cryo-EM approaches as described in our funded RF1NS133651 grant to pursue these currently unfunded objectives as a supplemental project. This proposal aligns with the major goals of our parent award and addresses Component 1 in the NIH INCLUDE Project’s notice of special interest (NOT-OD-22-137). Successful outcomes from this supplemental work will contribute to the preparation of a new R01 application to the INCLUDE Project, in which we develop structure-guided molecular diagnostics to more precisely define the heterogeneous tau neuropathologies that arise across the lifespan in Down syndrome.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY / ABSTRACT Chronic pain is the leading cause of disability worldwide, yet effective treatments remain elusive for most chronic pain syndromes. Chronic neuropathic pain, resulting from a lesion or disease affecting the nervous system, produces central sensitization and maladaptive plastic changes across distributed pain processing circuits. Consequently, there is an urgent need to understand how targeted brain stimulation can influence these widespread pain networks. My overarching career goal is to become a clinician-investigator with an independent computational neuroscience laboratory dedicated to studying relationships between brain network organization and chronic pain. I aim to leverage this knowledge to develop safer and more effective personalized treatments for chronic pain. The K23 award will provide critical support, mentorship, and training in 4 areas crucial to the success of my future independent research program, including: (1) advanced functional magnetic resonance imaging (fMRI), network neuroscience, and machine learning methods; (2) non-invasive brain stimulation with repetitive transcranial magnetic stimulation (rTMS); (3) clinical trial design, management, and statistics; and (4) experimental pain research. The proposed research applies this training to fMRI and multimodal pain outcomes data collected from ongoing trials of rTMS at the primary motor cortex (M1), provided as a therapy for chronic neuropathic pain. Although M1 is an established rTMS target for pain, clinical outcomes are variable and unpredictable. I hypothesize that chronic pain results from hyperconnected brain networks and that targeting highly connected network hubs within M1 can improve network function and reduce pain. To test this hypothesis, I will first perform a retrospective analysis of data from a pilot clinical trial comparing high frequency excitatory and low frequency inhibitory rTMS at M1 to examine the relationship between pain relief and the "hub-ness" of the stimulated target (Aim 1). I will then test whether pain-relieving high-frequency M1 rTMS reduces pathologically hyperconnected global brain networks (Aim 2). Finally, to prospectively validate network hub stimulation as a pain-relieving intervention, I propose a new, randomized, controlled cross-over trial comparing high frequency rTMS at personalized hubs versus non-hubs within M1 (Aim 3). Successful completion of this study will identify and prospectively validate regional and global network characteristics associated with pain relief from M1 rTMS. Planned analyses will establish tools and methods capable of predicting new, personalized rTMS targets in future patients with chronic pain. Results will be used in NIH R01 and HEAL grant applications for funding to support the ongoing development and implementation of personalized pain-relieving interventions, to provide much needed alternatives to opioid-based analgesia. Collectively, this training and research will position me as a global leader in the development of personalized, non-invasive, brain-based therapies for pain.

NIH Research Projects · FY 2026 · 2025-08

7. PROJECT SUMMARY The proposed work focuses on interorgan communication between the gut microbiome and the joint in the pathogenesis of osteoarthritis (OA). Although historically viewed as a disease caused by mechanical overload, recent studies indicate that systemic factors are a major contributor to OA. The gut microbiome is a critical regulator of systemic inflammation and is readily modified in individuals, making it an enticing therapeutic target. Studies by our group and others indicate that the gut microbiome is a potent regulator of OA capable of enhancing or blunting trauma-induced osteoarthritis in animal models. Aging is associated with alterations in the composition of the gut microbiota, increased systemic inflammation and increased susceptibility to OA. In several cases, age-related changes in the gut microbiome are major contributors to disease. However, it is not known if the gut microbiome influences spontaneous age-related osteoarthritis, nor is it clear if age-related changes in OA are due to age-related changes in host physiology or age-related changes in the gut microbiota. Clinical and preclinical studies have implicated microbially-derived molecules that transit from the gut into the systemic circulation as regulators of the relationship between the gut microbiome and the joint. However, the association between microbiome-induced changes in musculoskeletal tissues and microbially-derived metabolites is currently limited to examination of fecal samples. Supported by robust PRELIMINARY STUDIES suggesting an effect of the gut microbiome on age-related OA and implicating key microbiota-dependent metabolites with severity of OA, we propose the global hypothesis that the gut microbiota regulates the effects of aging on joint degeneration. Specifically, we ask: Does the microbiome influence spontaneous, age-related joint degeneration? To what degree do age-related changes in the gut microbiota v. age-related changes in host physiology promote joint degeneration? Which of the circulating microbiota-dependent molecules link the gut microbiome to joint degeneration? How do these effects differ by sex? The proposed work has three aims explored using mouse models: 1) Determine the effects of the gut microbiome on age-related spontaneous osteoarthritis; 2) Determine the contribution of age-related changes in the gut microbiota to osteoarthritis; and 3) Determine the role of circulating microbiome-derived metabolites on the severity of osteoarthritis. At the completion of the proposed work, we will have addressed key interactions between the gut microbiome and aging in the context of joint degeneration and determined the effects of microbiota-derived molecules as signaling factors linking the gut microbiome to OA.

NIH Research Projects · FY 2025 · 2025-08

Contact P.I.: Ostrem, B.E.L. PROJECT SUMMARY Preterm white matter injury (PWMI) is the most common type of brain injury in premature infants and is associated with adverse neurological outcomes, including cerebral palsy, cognitive and learning disabilities, and seizures. PWMI is caused by a differentiation arrest in the oligodendrocyte (OL) lineage, and a failure of myelination. As there are no specific treatments, care is supportive and focused on rehabilitation. However, novel high-throughput screening methods have enabled the identification of remyelinating, or pro-myelinating, compounds that promote OL differentiation and myelination. We hypothesize based on overlapping mechanisms of remyelination and developmental myelination that remyelinating compounds are an untapped source of potential therapeutics for PWMI. Here, we demonstrate the development of a therapeutic pipeline for PWMI where remyelinating compounds are identified using a high-throughput in vitro screening platform, binary indicant for myelination using micropillar arrays (BIMA). Screen hits that meet key criteria suggestive of safety and feasibility in neonatal neurological applications are validated in vitro and tested in a murine chronic hypoxia model of PWMI. We further propose to complete a Phase I clinical trial of a promising remyelinating compound that has completed testing through our preclinical pipeline, clemastine, in neonates with PWMI. This oral antihistamine is approved in patients ages 3 and older for allergic indications and was recently shown to promote remyelination in adults with multiple sclerosis. Neonatal safety and pharmacokinetic (PK) data are needed prior to larger efficacy studies for PWMI. We will perform a Phase I/Ib dose expansion/dose escalation study to test the safety and PK of clemastine in preterm neonates with PWMI. We have worked closely with the FDA, expert collaborators, and parent advocates on an adaptive, PK-guided trial design that accounts for prior pitfalls in clinical trials for newborn brain disorders. We will enroll 20 neonates born at ≤32 weeks gestation, who have PWMI identified by neuroimaging. Neonates will be treated with clemastine for 14 days while undergoing close monitoring and blood sampling for PK measurements in the intensive care nursery. Blood and imaging biomarkers will be obtained at enrollment and 3 months after treatment and neurodevelopmental outcomes will be followed for 2.5 years on all participants. This trial will inform the dosing for and design of a randomized, placebo-controlled, Phase II trial of clemastine for PWMI. Overall, our high-throughput drug discovery platform combined with our neonatal feasibility and prioritization system is a new and promising approach to therapeutic development for PWMI. Our trial design will serve as a road map for similar future trials for conditions affecting the newborn brain. The application of remyelinating therapies to neonatal brain injury is conceptually innovative, and our preliminary data are highly supportive of the potential success of this approach. This work has the potential to fundamentally change the therapeutic landscape, and approach to drug development, for a devastating neonatal neurological problem with no current treatments.

NIH Research Projects · FY 2025 · 2025-08

Project Summary/Abstract Research on psychedelics, substance use (SU), and mental health (MH) has shown promising results in the treatment of depression, anxiety, post-traumatic stress disorder, alcohol use disorder, and smoking cessation. Psychedelics can promote the development of new neural pathways, which may lead to improved health outcomes. Psychedelic-assisted therapy (PAT) involves the professionally supervised use of psychedelics as part of a psychotherapy program and offers a paradigm shift for more effective and safer options for SU and MH treatment. PAT could improve the lives of people with HIV (PWH) who experience SU or MH challenges at rates higher than the general population. These SU and MH challenges are strongly associated with lower medication adherence, reduced virologic suppression, disease progression, and high mortality among PWH. Treatment of SU and MH concerns can reverse adverse health effects, lead to improved clinical outcomes, and prevent onward HIV transmission. Therefore, parallel to the HIV treatment cascade, there is an urgency to address and achieve global treatment goals for SU and MH challenges. However, the low representation of minority groups among health researchers undermines the ability of public health interventions to address these health disparities. There is an urgent need for more diversity in the HIV research community, and building a pipeline of researchers from diverse backgrounds is critical to advancing science. In this K24, Dr. Parya Saberi proposes a plan to (1) provide mentorship to early career trainees from diverse backgrounds; (2) provide opportunities to develop her knowledge and skills in research related to SU, MH, PAT, and implementation science; and (3) provide preliminary data to inform subsequent patient-oriented research proposals by her and her mentees to address equity in SU or MH treatment and PAT among PWH. Therefore, Dr. Saberi’s mentoring goals include increasing the number of her mentees, enhancing her mentoring competencies, and providing a training program for mentees from diverse backgrounds to launch and support their careers. Mentees will leverage the infrastructure and resources of Dr. Saberi’s ongoing research and her collaborations with multidisciplinary researchers in the areas of HIV, SU, MH, PAT, and implementation science. The research aims of the proposed K24 are: (1) to assess the interests, concerns, motivators, and deterrents of using PAT among PWH experiencing moderate to severe SU or MH challenges and (2) to examine priorities for the research agenda and future clinical implementation of PAT among PWH experiencing SU or MH challenges. Therefore, funding from this K24 will allow Dr. Saberi to grow a structured program of training for researchers from diverse backgrounds and to conduct research that addresses the need for effective SU and MH interventions, enhanced scientific rigor in PAT research, and increased research to improve health equity for SU and MH treatment among PWH.

NIH Research Projects · FY 2026 · 2025-08

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. Adolescence is an important period for both social media exposure and substance use experimentation. By age 18, 84% of adolescents are social media users, and emerging evidence suggests that social media use may contribute to substance use. However, the mechanisms linking social media and substance use remain unclear, with significant knowledge gaps around patterns of use, sensitive developmental windows, and protective parenting strategies. This project aims to address these gaps using data from the Adolescent Brain Cognitive Development (ABCD) Study, a large longitudinal cohort of 11,875 adolescents followed for over 10 years. We will explore prospective associations between social media use and substance use (cannabis, nicotine, alcohol). Our long-term goal is to provide adolescents, parents, clinicians, and policymakers with guidance on the use of social media to reduce early substance use. Our central hypothesis is that social media use is prospectively associated with early substance use through problematic social media use patterns (measured by the Social Media Addiction Questionnaire) and positive substance use expectancies (favorable beliefs about substance use), while, in contrast, media parenting rules/monitoring can be protective. We propose the following specific aims: Aim 1: Determine longitudinal associations and sensitive windows linking social media use with substance use. Aim 2: Identify mechanisms linking social media use and substance use. Aim 3: Identify media parenting rules and monitoring that protect from substance use. To accomplish these aims, we will leverage comprehensive assessments of social media use and substance use among a large national prospective cohort, beginning in early adolescence in the ABCD Study followed annually for 10 years. The cohort uniquely starts before adolescence to capture the onset of social media use and substance experimentation from early to late adolescence, spanning important developmental periods. We will use robust longitudinal and structural equation modeling methods to analyze all available years of existing data in the ABCD Study. We will disseminate and translate findings to inform guidance for adolescents, parents, clinicians, and policymakers.

NIH Research Projects · FY 2025 · 2025-08

Abstract Among the estimated 155 million TB survivors worldwide, a substantial proportion have post-tuberculosis lung disease (PTLD), characterized by persistent respiratory complaints, abnormal lung function, and/or chest X-ray abnormalities. Despite its prevalence, PTLD is poorly characterized and poorly understood. Our objectives are to establish a well-characterized PTLD cohort in a low-income and high-TB/HIV burden country and determine whether sex, HIV serostatus, and air pollution are associated with distinct lung function-chest CT PTLD phenotypes. This proposal builds upon a successful 17+ year collaboration in Kampala, Uganda. Our previous research found significant sex-based and HIV-specific differences in spirometry post-pneumonia, and preliminary data indicate similar trends in spirometry, total lung capacity (TLC), and diffusing capacity for carbon monoxide (DLco) post-TB. Specifically, post-TB women with HIV exhibited stronger trends towards abnormal spirometry, TLC, and DLco compared to post-TB men with HIV; this was not seen in post-TB persons without HIV or in persons without TB. Post-TB women with HIV also had significantly higher PM2.5 levels than their male counterparts. These findings support our central hypothesis that there are sex-based and HIV-specific differences in the lung function-chest CT presentations of PTLD and that women with HIV are at increased risk. We hypothesize that these sex and HIV differences are exacerbated by higher PM2.5 levels in women and an enhanced deleterious effect of PM2.5 exposure in HIV. To address this, we will enroll 650 post-TB and 165 non- TB adults. We will determine the impact of sex and HIV on lung function-CT findings and investigate whether sex or HIV modifies the associations between PM2.5 and PTLD. Aim 1: Determine the impact of sex and HIV on spirometry, TLC, and DLco in persons with and without TB at baseline (end of TB treatment for TB) and follow- up (2 years later). Hypothesis: Post-TB women with HIV will have higher odds of abnormal lung function than post-TB men with HIV; no such sex difference will be found in post-TB persons without HIV (primary hypothesis) or those without TB (secondary). Aim 2: Determine the impact of sex and HIV on CT findings using conventional chest radiologist interpretation and novel, automated AI algorithms in persons with and without TB at baseline and follow-up. Hypothesis: Post-TB women with HIV will have higher odds of emphysema, fibrosis, small airways disease, and vascular abnormalities on CT than post-TB men with HIV; no such sex difference will be found in post-TB persons without HIV or those without TB. Aim 3: Determine whether sex or HIV are effect modifiers of the relationship between PM2.5 and lung function-CT findings using short- and long-term PM2.5 measurements from personal samplers and residential satellite-based spatiotemporal models. Hypothesis: The relationships between PM2.5 and PTLD findings will be more pronounced among women compared to men and among persons with HIV compared to persons without HIV. This research will serve as a foundational study of PTLD, leading to advancements in its diagnosis and management, mechanistic studies, and targeted interventions in the future.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT This proposal entails a five-year mentored career development plan that studies mechanisms of tau aggregation in Alzheimer's disease and frontotemporal lobar degeneration. This proposal includes technical training in iPSC-derived neuron cultures, CRISPR functional genomics and bioinformatics approaches, and in- depth neuropathologic analysis. The training plan also incorporates skills required to maintain an independent research group, including scientific communication, grant writing, mentorship skills, and lab management strategies. To achieve these goals, the candidate, Dr. Sarah Kaufman, has assembled a multidisciplinary mentorship team that includes Dr. Martin Kampmann (mentor), Dr. Bill Seeley (co-mentor), and advisory members Dr. Bruce Miller and Dr. Jason Gestwicki. Together, this proposal will allow her to establish a rigorous independent laboratory that combines her previous graduate training, clinical interest in Alzheimer's disease and frontotemporal dementia, and scientific expertise in tauopathies to become a leader in neurodegenerative disease research. Tauopathies including Alzheimer's disease (AD) and frontotemporal lobar degeneration with tau inclusions (FTLD-tau) are devastating neurodegenerative diseases that currently affect over 30 million people worldwide. Tau aggregates in AD and FTLD-tau adopt unique, disease-specific conformations, or “strains,” that deposit in selectively vulnerable neurons. Different tau strains are now believed to underlie the different disease phenotypes and atrophy patterns observed across tauopathies. However, the mechanisms that give rise to different tau strains and their link to selective vulnerability and neuronal loss are not known. Several human induced pluripotent stem cell (iPSC) derived excitatory neuron (iNeuron) models have been developed to study tauopathies. However, these models do not readily accumulate tau aggregates, which has hindered their use when studying mechanisms of tau strains or selective vulnerability. This proposal describes the creation of a novel iNeuron model that stably maintains full-length tau aggregates seeded from human tauopathies that are associated with distinct tau strains. An initial large-scale CRISPRi screen identified several novel regulators of tau aggregation in this iNeuron model, including the lesser-known ubiquitin-like UFMylation pathway that plays an important role in protein homeostasis, and several RNA binding proteins known to directly interact with tau protein. Aim 1 of this proposal will elucidate the role of RNA binding proteins in tau aggregation and selective vulnerability across different tau strains, followed by analysis in human tauopathy subjects. Aim 2 will elucidate the mechanisms by which UFMylation modulates tau aggregation in neurons. Together, the proposed research will provide important insight into the mechanisms that underlie tau aggregation and the link between tau strains and different patterns of selective vulnerability observed across tauopathies.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY / ABSTRACT I am an MD/PhD-trained neurointensivist and physiologist navigating a critical career development transition towards becoming an independent investigator. My long-term goal is to improve clinical outcomes of patients with severe ischemic strokes admitted to the intensive care unit (ICU). In the ICU, we can change the levels of carbon dioxide (CO2) or hydrogen ion (H+) in the bloodstream of intubated, mechanically ventilated patients. However, we do not know the optimal ranges of H+ or CO2 to target to support brain recovery and limit secondary injury following stroke. Experimentally elevating systemic CO2 and H+ can reduce the size of strokes in rodents, by mechanisms that are incompletely understood. H+ and CO2 influence cerebral blood flow, and may also shift cellular energy phenotypes in neurons or other brain cells. In parallel, H+ and CO2 levels may also inhibit spreading depolarizations (SDs), waves of powerful neuronal activation that propagate through the vulnerable brain territories adjacent to infarcts, exert immense energetic stress on neurons, and kill neurons with insufficient metabolic supply. SDs play a key role in expanding the size of strokes for many days, corresponding to the time period when patients are in the ICU. In this project, my objective is to identify protective and harmful mechanisms of acid-base physiology relevant to stroke. My hypothesis is that augmenting H+ will improve metabolic supply and energy utilization, inhibit SDs, and reduce the size of strokes. I will test this hypothesis with two specific aims: 1) comparing the effects of systemic and cerebral H+ vs CO2 on cerebral blood flow, cellular energy metabolism, and stroke outcomes in mice, and 2) testing whether elevated extracellular H+ decreases the incidence of SD by inhibiting NMDA receptors. The significance of the proposed research is that completing these studies will provide high-quality physiologic evidence to inform clinical guidelines for the management of acid-base status in ICU patients with stroke and other acute brain injuries.

NIH Research Projects · FY 2026 · 2025-08

Project Summary/Abstract Idiopathic pulmonary fibrosis (IPF) is a devastating and progressive lung disease characterized by extensive fibrotic remodeling leading to respiratory failure and death. Despite recent advancements, no curative treatments exist for IPF, underscoring the urgent need to identify novel therapeutic targets. Recent studies suggest a potential role for purinergic signaling in IPF pathogenesis, particularly through the actions of extracellular ATP (eATP): Elevated eATP levels have been observed in the bronchoalveolar lavage fluid of IPF patients, implicating this signaling molecule as a potential driver of disease progression. However, the functional impact of eATP in the context of lung fibrosis remains poorly understood. Addressing this knowledge gap, our research has identified the fibroblast-specific ATP receptor P2rx4 as a key mediator of fibrotic signaling. Deletion of P2rx4 in fibroblasts has been shown to reduce lung fibrosis in bleomycin-treated mice, making it a promising target for therapeutic intervention. Notably, P2rx4 is highly expressed in pathologic fibroblasts in both human and mouse models of lung fibrosis, reinforcing its relevance in the fibrotic process. Our preliminary data suggest that the eATP-P2rx4 signaling axis regulates several profibrotic fibroblast phenotypes, including the upregulation of collagen genes, production of the cytokine IL-6, and enhanced fibroblast proliferation. This proposal seeks to dissect the underlying mechanisms by which P2rx4 influences these fibrotic pathways in fibroblasts. In Aim 1, we will test the hypothesis that the eATP-P2rx4-p38 MAP kinase signaling pathway is a critical regulator of fibroblast activation and the emergence of pathologic fibroblasts. We will also investigate the role of mitochondrial reactive oxygen species (mitoROS) in mediating P2rx4-dependent p38 MAP kinase activation. Aim 2 will focus on evaluating the therapeutic potential of targeting P2rx4. P2rx4 inhibition will be tested in both translationally relevant mouse models of lung fibrosis and in precision-cut lung slices derived from IPF patient explants to assess efficacy in reducing collagen deposition and fibrosis. Aim 2 will also test the relevance of the profibrotic eATP-P2rx4-p38 signaling that our preliminary data indicate for mouse in primary human lung fibroblasts from healthy donor and IPF lung explants, by siRNA knockdown of P2rx4 and p38. Taken together, these studies will transform understanding of purinergic signaling in lung fibrosis, providing critical insights into the molecular mechanisms that drive fibroblast activation IPF. By identifying and validating ATP-P2rx4 signaling as a novel therapeutic target, this research has the potential to pave the way for the development of new and effective treatments for IPF.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT Type 1 diabetes (T1D) is a chronic autoimmune disease characterized by hyperglycemia due to progressive immune-mediated destruction of pancreatic beta cells. Recent work from our laboratories has shown that hyperactivation of the unfolded protein response (UPR) to ER stress in the immune-targeted beta cells may be a critical early event in the development of T1D. We have developed novel pharmacological reagents called ‘PAIR’s that allow us to manipulate components of the UPR. Importantly, first-generation versions of small molecules delivered to NOD mice can efficaciously prevent and even reverse diabetes in this T1D model. Thus, in this collaborative grant we will capitalize on the complementary expertise of the investigators to optimize these next-generation “PAIR” molecules for potency and selectivity, conduct proof-of-concept studies using human islets (ex vivo and in transplanted mice), and perform key enabling steps needed to advance these candidates further into the clinic for treating human patients with T1D.

NIH Research Projects · FY 2025 · 2025-08

Project Summary Our earlier work identified the neural stem cells (NSC) and intermediate progenitors that generate new neurons and glial cells in the adult mouse brain. These NSCs are regionally specified to produce unique sets of inhibitory interneurons that migrate along the rostral migratory stream (RMS) to the olfactory bulb. Unlike rodents, the adult human brain does not contain an RMS, but this and other migratory streams are present in infants. In young children, we recently found two major streams of migrating young neurons (the ARC and the EC stream) that supply inhibitory cortical interneurons (cIN) to the frontal and entorhinal cortices respectively. The neuronal progenitor cells that generate these postnatally recruited young neurons have not been identified. Our overall goal for the next 8 years is to investigate how NSC niches transition from embryonic to postnatal stages in the human forebrain. We aim to characterize the different NSCs and progenitor populations involved in the production of large numbers of cINs required for the greatly expanded human neocortex. We plan to focus on the ventral forebrain germinal regions, in particular, on the caudal ganglionic eminence (CGE), which is the source of half of all cINs in humans, including the majority of cells observed in the ARC and the EC stream. CGE-derived cINs are considered key to higher cognitive function, and dysregulation of their numbers has been linked to neurological disorders and tumor formation. We hypothesize that the human CGE has increased its output of cINs by (1) increasing the proliferation of intermediate progenitors at multiple stages in the cIN lineage, and (2) extending the period of neurogenesis into postnatal life. We propose to use multi-omics, spatial transcriptomics, and electron and light microscopy to investigate the cellular composition and organization of the human CGE. We aim to identify the different CGE progenitor populations and how this germinal niche is organized. Preliminary data suggests that epidermal growth factor receptor (EGFR) could be key to CGE intermediate progenitor amplification. Using CGE organoids and genetic approaches in mice, we will investigate the role of EGFR signaling in CGE neurogenesis and gliogenesis. We will then study what progenitor cells persist postnatally in the CGE and other ganglionic eminences and how their niches change from prenatal to postnatal stages. Finally, we propose to resolve the controversy of whether NSCs in the postnatal brain produce both neurons and glial cells in vivo. The proposed work will elucidate how cIN production is amplified to satisfy the needs for a greatly expanded human neocortex. This basic new knowledge of human brain development could lead to novel approaches for the treatment of neurological disorders and the identification of progenitor cells implicated in tumor formation.

NIH Research Projects · FY 2025 · 2025-08

Abstract This research project aims to provide a step-change in our comprehension of cardiac pacemaking and sinus node dysfunction (SND) by introducing the concept of “conditional pacemaker cells” within anatomically and functionally distinct “specialized pacemaker cell subpopulations (SPCS)”. SND, which disrupts the heart’s natural pacing, is linked to significant cardiovascular morbidity and mortality, posing a global health challenge. SND manifests in various types, reflecting its complex and multidimensional etiologies. Despite the critical need for effective treatments, currently, no therapy is available to prevent or reserve the primary cause of SND. Previous research has primarily focused on the conventional pacemaker cells typically defined by their inherent basal automaticity, overlooking the potential crucial role of cells that activate only under specific conditions – “conditional pacemaker cells”. Our preliminary studies link specific types of SND to these previously unrecognized “conditional pacemaker cells” rather than conventional pacemaker cells, underscoring the necessity of identifying and characterizing these cells for better recognition of SND pathology and the development of targeted therapeutic strategies. To achieve this, we will leverage a refined advanced high-resolution optical mapping technology to directly “visualize” the pacemaking activities within distinct specialized pacemaker cell subpopulations in real time, which robustly preserves the cellular integrity, viability and RNA quality, facilitating the systematic investigations of these cell subpopulations at multiple levels as well as the precise integration of the organ-wide functionality, macroscope innervation, and single-cell transcriptomics in same SPCS in situ – a first in the field. Utilizing genetic perturbation, we aim to ascertain the unique pacemaking mechanisms of distinct SPCS and uncover the subcellular mechanism of each specific type of SND that links to its corresponding SPCS in an aged rabbit model. This project transcends the traditional concept of SAN heterogeneity by extending its focus beyond cells solely exhibiting basal automaticity. It challenges the conventional limits of SAN transcriptomics, which limited to marker genes identified solely from basal beating cells in pacemaker cell identification and clustering, and failed to connect genetically heterogeneous cell populations with specific pacemaking functions, or anatomical regions, therefore, offering novel insights into the complex of SAN pacemaking and SND.

NIH Research Projects · FY 2025 · 2025-08

Statement of the Problem: Pulmonary fibrosis (PF) is a chronic lung disease characterized by tissue scarring and is common in interstitial lung diseases (ILDs). ILDs contribute significantly to global mortality and healthcare expenses. Some ILDs are considered idiopathic, but research indicates that exposure to inhalants like vapors, gases, dust, and fumes at work or in the environment can promote fibrosis and inflammation, leading to PF. Most research focuses on treatments and comorbidities, neglecting the impact of preventable occupational and environmental exposures. Additionally, marginalized communities with higher exposure rates are rarely studied. Investigating inhalational exposures can help shape environmental policies and workplace safety, preventing PF in affected communities and vulnerable workers. Specific Aims: I aim to use epidemiologic tools to assess the occupational and environmental risk factors and progression of fibrotic ILD, analyzing a large dataset, the California Electronic Death Registration System (Cal- EDRS), and an ongoing prospective cohort, the University of California San Francisco UCSF ILD Cohort Study. In Aim 1, I plan to systematically develop a new PF-specific job-exposure-matrix (PF-JEM) and measure the association between occupational exposure and PF in the Cal-EDRS. In Aim 2, I will assess the interaction of environmental and occupational exposures on the occurrence of PF in the Cal-EDRS, using air pollution data from the CalEnviroScreen and geospatial techniques. To achieve Aims 1 and 2, I will utilize a case-control study design in the Cal-EDRS to measure the association between occupational and environmental exposures using multiple predictor models and stratified analysis. Finally, in Aim 3, I will evaluate the interaction of environmental and occupational exposures on clinical outcomes of IPF in the UCSF ILD Cohort Study, including survival, using multivariable Cox proportional hazards analysis with stratification. Significance of the results. The Cal-EDRS and the UCSF ILD Cohort Study offer a unique opportunity to study the interplay of occupational and environmental exposures on PF utilizing state-of-the-art exposure assessment techniques of a JEM and residential address geocoding. This study is innovative through its linkage of a novel PF-JEM to both a rich dataset of vital statistics records and an ongoing cohort study to investigate the risks and clinical patient outcomes of PF due to inhalational exposures. Career Development: With the above proposal, Dr. Gandhi will achieve her training goals of 1) Epidemiologic occupational exposure assessment, 2) Advanced environmental exposure assessment using geospatial techniques, and 3) Survival analysis and clinical outcomes assessment of exposed patients with ILD. The K23 award will facilitate her transition to independence as a physician-scientist who utilizes innovative epidemiological methods and exposure metrics to develop a research program focusing on clinical outcomes of patients with occupational and environmental lung disease.

NIH Research Projects · FY 2026 · 2025-08

Project Summary/Abstract One of the hallmarks of severe fibrotic injuries of the lung is the appearance of metaplastic airway basal cells in the damaged alveoli. In patients with idiopathic pulmonary fibrosis (IPF), this pathognomonic feature is described as “bronchiolization,” which denotes the appearance of bronchus-like structures in the scarred alveoli that leads to cystic remodeling in the lung. In this proposal, we seek to define pro-reparative differentiation pathways in both the fibroblast and stem cell compartment that can be leveraged as therapy. Our preliminary data demonstrated the appearance of an alveolar fibroblast population during fibrotic injury resolution in mouse that expresses the extracellular hedgehog antagonist, HHIP, concurrent with a metaplastic basal cell-to-alveolar type 2 (AT2) cell differentiation trajectory in the alveoli undergoing repair. Furthermore, we generated a chimeric HHIP protein that demonstrated efficacy in promoting basal-to-AT2 differentiation in vivo. Utilizing a combination of novel genetic tools to trace and delete HHIP+ fibroblasts, xenotransplant model of human metaplastic basal cells, and a novel pharmacologic reagent made in our lab to target metaplastic basal cell differentiation, this proposal will test the central hypothesis that HHIP+ alveolar fibroblasts emerge after fibrotic injury to promote the restoration of alveolar progenitors from metaplastic basal cells. Finally, we will determine whether HHIP can be leveraged as pharmacotherapy to reverse pathogenic remodeling seen in IPF, and establish a humanized preclinical model to test the effect of therapeutics to reverse metaplastic remodeling in the fibrotic alveoli. Successful completion of this proposal will define a stem cell lineage trajectory that leads to reversal of remodeling seen in IPF, and provide preclinical studies to validate this therapeutic strategy.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT Nearly half of child deaths occur during the neonatal period, and 80% of those occur in babies with low birthweight. Although tremendous progress has been made towards reducing under-five mortality globally, declines in neonatal mortality lag behind those observed in older children. Low birthweight babies are at increased risk of poor outcomes compared to those who are term-appropriate for gestational age, including mortality, stunting, and growth failure. Recent evidence has demonstrated that the incidence of wasting and linear growth failure is highest between birth and 3 months of age, substantially earlier than previously thought. Interventions are urgently needed to improve outcomes in low birthweight babies and those who are underweight and wasted (collectively referred to as infants at risk of poor growth and development); however, these interventions must not interfere with breastfeeding and thus some well-established interventions used to treat or prevent malnutrition in older children cannot be considered. We recently demonstrated that biannual mass azithromycin distribution reduces all-cause childhood mortality by approximately 25% in infants aged 1-5 months, with stronger effects seen in underweight infants. This study did not include neonates due to the risk of infantile hypertrophic pyloric stenosis (IHPS) that has been hypothesized to be associated with macrolide use during early infancy. Recently, our study team documented only a single case of IHPS among 21,832 neonates enrolled in a trial of azithromycin versus placebo administered to neonates aged 8-27 days for prevention of infant mortality, documenting no major risk of IHPS associated with azithromycin. Here, we propose an individually randomized trial to test whether oral azithromycin improves nutritional outcomes and reduces infectious burden among neonates aged 1-27 days who are at risk of poor growth and development. The primary outcome will be weight-for-age Z-score at 6 months of age compared between arms. We anticipate that the results of this study will provide definitive evidence on azithromycin as an early intervention for neonates at risk of poor growth and development, who are at the highest risk of adverse outcomes.

NIH Research Projects · FY 2025 · 2025-08

ABSTRACT Transcranial low-intensity focused ultrasound (LIFU) is an emerging modality for noninvasive and targeted modulation of cortical and deep targets in the human brain. LIFU has enormous potential to realize a transformative treatment for myriad neurological and psychiatric disorders, but significant challenges exist for the clinical translation of LIFU neuromodulation protocols due to the unknown underlying neurophysiological mechanisms and the relatively large parameter space that remains poorly understood. Dissecting the mechanistic underpinnings of LIFU neuromodulation with systematic and carefully controlled experimental approaches is a critical step to facilitate the effective development of therapeutic protocols. Recent studies have established that LIFU elicits differential responses in diverse neuronal cell types depending on the pulsing parameters . However, dynamic circuit-level effects resulting from synaptic interactions between cell types have remained largely undisclosed. In this project, we will develop a systematic approach based on cutting-edge technologies to generate new fundamental knowledge on the underlying mechanisms of LIFU neuromodulation in specific cell types, focusing on circuit and network-level effects. In our analysis of the temporal dynamic responses, we will directly assess whether LIFU can enhance gamma-frequency (30-100 Hz) neural oscillations, which play a key role in information processing and cognition and are implicated in the pathophysiology of many neuropsychiatric disorders, including autism, schizophrenia, and neurodegenerative disorders. In all the proposed experiments, we will implement carefully designed control conditions to disentangle direct neuromodulatory outcomes from potential nonspecific auditory effects. In Aim 1, we will use cell-type specific fluorescent calcium indicators to map the LIFU neuromodulation parameter space in excitatory neurons and in parvalbumin (PV) and somatostatin (SST)-positive inhibitory interneurons. These cell types are particularly interesting due to their specialized role in the generation of gamma oscillations. In Aim 2, we will record cell- type-specific transmembrane voltage dynamics from cortical excitatory and inhibitory neurons to shed light on the circuit-level responses to LIFU neuromodulation. In Aim 3, we will use functional ultrasound imaging to map whole-brain network-level responses. Taken together, our Aims will yield new fundamental insights on the underlying mechanisms of LIFU neuromodulation. Excitingly, our Aims will lay the groundwork for a clinically translatable approach to systematically modulate neural oscillations in deep brain regions.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Calcific aortic stenosis, an age-related calcification and narrowing of the aortic valve, is the most common heart valve disease and is responsible for over 10,000 deaths and 90,000 heart surgeries each year in the United States. Risk for aortic stenosis is known to have an inherited component, and genetic syndromes such as familial hypercholesterolemia predispose to early disease onset. Nevertheless, trials of cholesterol- lowering medications did not slow disease progression, and at present there are no effective medical therapies for aortic stenosis. There is a critical need to identify risk factors for the development of aortic stenosis and to identify new therapeutic targets. We submit that studying normal population variation in aortic valve measurements (including valve area, mean gradient, and peak velocity) has the potential to address these challenges. Our preliminary data from over 40,000 people provide early evidence that there are shared factors—such as genetic variants near the gene encoding lipoprotein(a)—that are common both to normal variation in aortic valve measurements and to clinically identified aortic stenosis. The specific objective of this proposal is to quantify aortic valve measurements in 100,000 people and to deeply characterize their epidemiological and genetic bases. The overall hypothesis for the proposed work is that factors linked to normal variation in aortic valve hemodynamics in the population underlie the risk for the pathogenesis of aortic stenosis. In Aim #1 of this proposal, we will measure the aortic valve in 100,000 UK Biobank participants using our deep learning models; validate these measurements externally in clinical data from UCSF; and develop deep learning models to identify bicuspid aortic valves. In Aim #2, we will comprehensively characterize the epidemiologic, metabolomic, proteomic properties of aortic valve measurements as well as aortic stenosis as identified in the electronic health record. We will also develop deep learning models to estimate the aortic valve measurements from electrocardiographic data, and externally validate these models in data from tens of thousands of UCSF patients. In Aim #3, we will perform comprehensive analyses of the common genetic basis for variation in 100,000 people with aortic valve measurements and nearly 1.5 million people in a case/control analysis for aortic stenosis. We will also study contributions from rare pathogenic variants and clonal blood cells. The large-scale study of normal variation in aortic valve hemodynamics is an innovative approach for understanding the pathogenesis for aortic stenosis. We believe that our aims are consistent with the NHLBI’s mission of understanding the causes of disease and enabling translation of basic discoveries into clinical practice.

NIH Research Projects · FY 2025 · 2025-08

Project Summary Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB), is one of the leading causes of death due to infectious disease. Antibiotic treatment of Mtb is arduous and multidrug-resistant strains are on the rise, thus fueling the need for novel therapeutics. During infection, subpopulations of Mtb cells are phenotypically heterogeneous and can survive extensive chemical regimens, thus preventing the cure of disease. Cellular heterogeneity arises from the asymmetric growth and division of the bacterium, resulting in daughter cells with varying characteristics. As mycobacteria replicate, they must coordinate the division complex (divisome) and elongation machinery (elongasome) spatially and temporally for proper cell division. These protein systems share some similarities with the systems found in Escherichia coli and Bacillus subtilis, which grow and divide symmetrically. However, mycobacteria lack several major elongasome and divisome components that are found in these model bacteria, and components that are shared contain substantial N- and C-terminal expansions, which may interact with other co-factors. Immunoprecipitation and light microscopy suggest that the mycobacterial elongasome and divisome components potentially crosstalk with one another and form large multi- subunit protein complexes. PonA1 and CrgA are critical proteins of both the elongasome and divisome and are thought to make a multitude of interactions with other proteins in these systems. However, how these components coordinate and interact to support asymmetric growth and division is not well understood at a structural and mechanistic level. This proposal is focused on the biochemical and structural characterization of the elongasome and divisome from Mtb and the non-pathogenic model mycobacterium, Mycobacterium smegmatis. Bacterial genetics and endogenous protein purifications will be employed to purify PonA1, CrgA, and their associated complexes directly from mycobacterial cells, and single-particle cryo-electron microscopy will be used to determine their 3D structures (Aim 1). Novel proximity labeling will be utilized to define the protein- protein interaction networks of divisome and elongasome components and to decipher the roles of their N- and C-terminal expansions (Aim 2). Cutting-edge cryo-electron tomography will be implemented to capture 3D snapshots of dividing mycobacteria to determine how the elongasome and divisome proteins are structured and organized in situ (Aim 3). This proposal is innovative in that it applies emerging techniques to provide a more complete picture of the protein complexes that constitute the elongasome and divisome and how they interact to facilitate asymmetric growth and division in mycobacteria. This work will provide important insights into the structures of these multi-protein complexes that can be used for the design of new therapeutics for TB.

NIH Research Projects · FY 2025 · 2025-08

Project Summary/Abstract Understanding the contribution of menopause to clinical outcomes in HIV is an emerging health priority. There are over 20 million girls and women living with HIV (WWH) worldwide and over 50% of WWH in the US are experiencing the transition to menopause, a multi-year period characterized by declines in ovarian reserve that spans the premenopausal, perimenopausal, and postmenopausal phases. Perimenopause is a time of increased multi-morbidity risk, highlighting its importance as an optimal time for preventive interventions. HIV and menopause are independently associated with a higher burden of cardiovascular disease (CVD) and other comorbidities, but it is unclear whether HIV and menopause have synergistic effects. Studies of menopause in WWH have been impeded by the irregular menses and amenorrhea caused by chronic illness. As a result, it is unclear how menopause impacts CVD risk in WWH and whether perimenopause is a critical period for CVD prevention in WWH. This proposal leverages the NHLBI-funded MACS/WIHS Combined Cohort Study, which serially measures anti-Müllerian hormone, a biomarker of ovarian reserve, to precisely determine the pre-, peri- , and postmenopausal phases. The overall objective of this proposal is to determine the contribution of HIV and menopausal phase to CVD risk parameters (e.g., insulin resistance, lipids, hypertension) and subclinical atherosclerosis, and to examine the influences of immune activation and adiposity. The specific aims are (1) to determine the association of HIV and menopausal phase with CVD risk parameters; (2) to evaluate the association of HIV and menopausal phase with immune activation and whether immune activation mediates subclinical atherosclerosis using carotid imaging; and (3) to examine the association between MRI-measured adipose tissue volume and immune activation across the menopausal transition in WWH. Determining whether menopause and HIV have a synergistic effect on CVD risk could reveal mechanisms that drive CVD in WWH and identify therapeutic targets to prevent CVD. This K23 proposal also supports the career development of Dr. Rebecca Abelman, Assistant Professor of Medicine at the University of California San Francisco. Dr. Abelman’s long-term goal is to become an independent investigator studying ovarian aging and its role in comorbidity burden and organ dysfunction in WWH. Her short-term career goal is to examine the effect of menopausal phase on the development of cardiometabolic outcomes in WWH. To achieve her career goals, Dr. Abelman proposes training in (1) biomarker testing, analysis, and interpretation; (2) causal mediation and longitudinal data analysis; (3) imaging in patient-oriented studies of menopause and comorbidities; and (4) leading independent ancillary studies in large observational cohorts. To help guide her transition to career independence, Dr. Abelman has assembled a highly experienced mentorship team with deep expertise in HIV, menopause, the inflammatory consequences of HIV infection, biomedical imaging, and biostatistics.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT Background: Despite the success of T cell therapies in treating hematologic malignancies, solid tumors remain a significant challenge due to their dense fibrotic stroma, immunosuppressive microenvironments, and antigen heterogeneity. Current adaptive cell therapy approaches struggle to overcome these barriers, highlighting the need for novel strategies. Myeloid cells, including macrophages and dendritic cells, possess inherent abilities to infiltrate tumors, secrete cytokines, and cross-present antigens within the tumor microenvironment, making them ideal candidates for therapeutic engineering. However, engineering these cells is technically challenging, and tumors often co-opt myeloid cells to suppress adaptive immunity. We aim to overcome these obstacles by applying advanced genetic engineering tools to reprogram myeloid cells, enhancing their tumoricidal activities and resistance to polarization into immunosuppressive states. Toolbox: We have developed a toolbox of new techniques that will allow us to overcome key engineering challenges in human myeloid cells. Recognizing that nucleofection can lead to significant toxicity and dysfunction, we have found that using enveloped delivery vehicles (EDVs) to deliver Cas9 preserves myeloid cell viability and functionality. Additionally, through a collaboration with the Landau lab, we have shown that lentivirus incorporating the Vpx system leads to highly efficient transduction of human myeloid cells. We have further engineered these lentiviruses to express a mutant VSVG and scFvs targeting myeloid-specific surface proteins, enabling selective transduction of specific myeloid compartments. Approach: 1) CRISPR Gene Editing: These tools will enable us to perform CRISPR-based knockout and base editing screens in primary human macrophages and dendritic cells to identify and modify key genes that regulate myeloid cell polarization and effector functions. In addition, we will explore base editing of genes with known roles and mutations in autoinflammatory diseases driven by overactive myeloid cells. 2) Synthetic Receptor (Innate-CAR) Library: To complement these gene editing approaches, we will design and test a library of synthetic receptors, which we call Innate-CARs, to enhance myeloid cell immunogenicity. These Innate-CARs will combine a tumor- specific scFv with intracellular domains from a variety of diverse innate immune receptors and intracellular proximal signaling molecules to optimize myeloid cell responses to tumor antigens. After we screen and identify top-performing Innate-CARs and gene edits, we will use a variety of immunodeficient and immunocompetent preclinical models to assess these enhancements in CAR-myeloid cell antitumor immunity. 3) In Vivo Engineering: Using our novel myeloid-targeting lentiviruses, we will evaluate the efficiency and efficacy of in vivo manufactured Innate-CAR myeloid cells in tumor-bearing humanized mouse models. Impact: By exploring the synthetic space of myeloid cell biology, we aim to push the boundaries of current immunotherapy paradigms and pave the way for new, effective living medicines to treat refractory solid tumors.

NIH Research Projects · FY 2025 · 2025-08

Project Summary/Abstract This project seeks to develop and optimize chimeric antigen receptor (CAR) T-cell therapies for acute myeloid leukemia (AML). AML is the most prevalent acute leukemia in adults, yet has a dismal 5-year survival rate un- der 30% with current standard-of-care therapies. Despite their well-documented efficacy in the treatment of other hematologic malignancies, CAR-T therapies have shown limited efficacy in treating AML, highlighting the need for additional innovative research in this area. The most well-studied AML CARs in development use single-chain variable fragments (scFvs) to target CD33, CD123, or CLEC12A, resulting in severe toxicities in patients due to their concurrent expression on hematopoietic stem and progenitor cells (HSPCs). Instead, this study proposes to target CD70, a marker highly expressed on AML tumor cells and leukemic stem cells but not on HSPCs, using a “natural ligand” CAR-T approach, based on CD70’s unique native receptor, CD27. This approach has shown improved tumor cytotoxicity, CAR-T proliferation, and survival in preclinical models compared to scFv-based anti- CD70 CAR-Ts. Given their superior safety profile, CD27-based CAR-Ts merit further optimization for efficacy. Furthermore, there have been no comprehensive attempts to produce variants of natural ligand CARs with al- tered binding affinity and, thus, potentially improved CAR-T efficacy. Therefore, I hypothesize that by employing structure-based deep learning algorithms and high-throughput screening techniques, I can identify coding se- quence alterations in CD27-based CARs that enhance their efficacy without increasing off-tumor toxicity. In Specific Aim 1, I will evaluate the in vitro and in vivo efficacy of a set of CD27-variant CARs with computation- ally predicted alterations in affinity to CD70. The phenotypic differences of these CD27-variant CARs, particularly those influencing tumor cytotoxicity, will also be analyzed to elucidate the principles guiding natural ligand CAR- T function. In Specific Aim 2, I will perform an in vivo competition assay of CAR-T proliferation to screen a large library of CD27-variant CARs. The leading candidates will undergo further validation for tumor cytotoxicity, sur- vival benefits, and CAR-T phenotypes, along with verification that the low off-tumor toxicity of the CD27-based CAR is maintained. Successful execution of these aims is expected to yield superior CD27-based CAR designs that would be beneficial in various AML scenarios and potentially other CD70-expressing malignancies, with the long-term potential for translation to the clinic. Additionally, this natural ligand CAR development pipeline could be extended to additional cancer antigens, signifying a broad therapeutic potential. I will lead this project under the sponsorship of Dr. Arun Wiita, a physician scientist and expert in hematological malignancies and cancer antigen identification, and the co-sponsorship of Dr. Justin Eyquem, an expert in CAR- T development and CAR screening technologies. Completion of my research plan, alongside my comprehensive training plan and robust mentorship from my research environment, will strongly equip me to become an inde- pendent physician scientist focused on cancer immunology and immunotherapy research.

NIH Research Projects · FY 2026 · 2025-08

PROJECT SUMMARY/ABSTRACT This application for a K23 career development award for Charles Windon, MD, will support his long-term goal of becoming an independent clinical researcher. Dr. Windon is a behavioral neurologist and Assistant Professor in the Department of Neurology at the University of California, San Francisco, within the Memory and Aging Center (UCSF MAC). He seeks to become an independent investigator in the field of biomarker- informed inequities research and address inequities in Alzheimer’s Disease and Related Dementias (AD/ADRD). The support provided by this K23 will allow Dr. Windon to achieve the following training goals (TG): TG1: knowledge in PET imaging analysis and plasma biomarker analysis; TG2: advanced statistical methodology knowledge for high complexity imaging and clinical data; TG3: skill building in analytic approaches to individual- and neighborhood-level SDOH; and TG4: grantsmanship, dissemination of clinical research, and other areas of professional development. He will incorporate the findings of his K23 into an R01 that develops and explores models validated in larger cohorts more richly characterized by SDOH measures and Alzheimer’s Disease and related dementias (AD/ADRD) biomarkers to best identify which pathological contributions to cognitive impairment should be emphasized in therapy for ethnoculturally diverse individuals. Dr. Gil Rabinovici, a world recognized leader in amyloid PET and fluid AD/ADRD biomarkers, will serve as primary mentor for this K23. He is joined by co-mentor Dr. Jacqueline Torres, a social epidemiologist who has extensive expertise in advanced epidemiologic and econometric methods to examine social determinants and inequities in AD/ADRD. Dr. Adam Boxer, an experienced researcher and clinical trials leader with expertise in blood-based biomarkers of AD/ADRD, Dr. Isabel Allen, an experienced professor with expertise in statistical analysis, and Dr. Amy Kind, a leading expert in neighborhood level social determinants, are collaborators. Dr. Windon will investigate in this proposal how global β-amyloid burden, measured by PET, and blood-based phosphorylated tau (p-tau) 217 and Neurofilament light (NfL) biomarkers are independently associated with AD/ADRD among diverse populations with varying social determinants of health (SDOH) profiles. Analysis of >2900 cognitively impaired diverse individuals with these biomarkers will be used for specific aims of: 1) Understanding differences in clinical presentation, dementia risk factors, SDOH profile, and rates of amyloid PET positivity between cognitively impaired Black/African American and Latinx individuals and Non-Latinx White individuals; 2) Understanding differences in β-amyloid burden by PET centiloid value between cognitively impaired Black/African American and Latinx and Non-Latinx White individuals and quantify relationships between p-tau 217 and NfL, respectively, and cognitive impairment among these groups; 3) Testing whether individual-level SDOH or neighborhood-level SDOH act as effect modifiers in the relationship between p-tau 217 and NfL and PET centiloid value, respectively, and cognition.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY The World Health Organization (WHO) has historically graded meningiomas according to histological features, and many WHO grade 1 meningiomas can be effectively treated with surgery or radiotherapy, but many WHO grade 2 or grade 3 meningiomas are resistant to treatment and cause significant neurological morbidity and mortality. Approximately 30% of WHO grade 1 meningioma develop recurrences that cannot be predicted from histological features, and some WHO grade 2 or grade 3 meningiomas are unexpectedly well controlled with surgery and radiotherapy. These data indicate that improvements in risk stratification and new therapies for patients with meningiomas are needed, but limited understanding of meningioma biology and the misconception that all meningiomas are benign has encumbered medical and scientific advances for patients with meningiomas. Using multiplatform molecular profiling on 1856 meningiomas from 12 institutions across 3 continents, we discovered a 34-gene expression biomarker that improve risk stratification compared to all other classification systems across all WHO grades. Moreover, our data demonstrate that gene expression profiling predicts which meningiomas will benefit from postoperative radiotherapy (32%). Cooperative group trials, clinical guidelines, and classification systems that incorporate our gene expression biomarker are under development. More broadly, the development of this biomarker provides a framework for understanding how meningiomas grow and respond to therapy, but it is unclear if the 34 genes that comprise this biomarker are drivers or merely markers of meningioma biology. Here we propose mechanistic and functional genomic interrogation of meningioma radiotherapy response genes using traditional mechanistic and function approaches (Aim 1) and a novel technique that links genetic and therapeutic perturbations to genomic phenotypes in single cells (Aim 2) across a unique biobank of meningioma cell lines that represent each meningioma DNA methylation group in humans. Our central hypothesis that the genes comprising our predictive biomarker of meningioma outcomes in humans represent therapeutic vulnerabilities that could enhance radiotherapy responses. Our studies will use genetic loss-of-function and rescue approaches alongside proliferation, clonogenic growth, and apoptosis assays (Aim 1) and simultaneous single-nuclei ATAC, RNA, and CRISPRi perturbation sequencing (snARC- seq) to perform mechanistic and functional genomic interrogation of meningioma radiotherapy response genes. To do so, we have assembled a biobank of 27 patient-derived meningioma cell lines that represent all DNA methylation groups in humans and completed a genome-wide CRISPRi screen to shed light on which genes from our 34-gene expression biomarker underly meningioma growth or response to radiotherapy. Successful completion of this project will determine if the 34 genes comprising our predictive biomarker are drivers of meningioma biology and will elucidate targets for future pharmacological studies in preclinical xenograft models and early phase clinical trials in patients.