University Of California, San Francisco

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY Low back pain is the leading cause of disability and is closely linked to disc degeneration. A key factor in the etiology of disc degeneration, and a critical obstacle to disc regeneration, is inadequate permeability of the cartilage endplates (CEPs) to vital nutrients that are needed to sustain the nucleus pulposus (NP) cells. Historically, low CEP permeability was attributed to matrix calcification. However, emerging data show that impermeable CEPs which block nutrient transport and associate with severer disc degeneration have higher amounts of collagen and aggrecan than permeable CEPs. Moreover, we found that ex situ treatment of the CEP with enzymes that selectively reduce collagen or aggrecan increases nutrient transport and improves NP cell survival. In this new project, we propose studies in CEP explants and whole-disc organ cultures that will: 1) reveal which CEP matrix constituent(s), i.e., collagen, aggrecan, and/or mineral, should ideally be reduced to increase CEP permeability while preserving overall CEP function; 2) discover how CEP treatments that target the various matrix constituents affect CEP and NP cell biologic response and matrix turnover; and 3) determine how meaningful are the effects of different matrix-modifying treatments on disc nutrition and NP cell function. Three complementary aims are proposed. In Aim 1 we will develop a relationship between CEP matrix modification, size-selective solute filtration, and material properties. Using permeation and mechanical tests, we’ll measure solute filtration properties and damage resistance of human CEP tissues treated with enzymes that either reduce collagen (full-length MMP-8) or aggrecan (truncated MMP-8, “tMMP-8”) alone and in combination with a treatment that reduces calcification (citric acid). We’ll test how targeting the various matrix constituents enhances glucose transport while retaining damage resistance and preserving the filtration of larger matrix fragments that are needed for proper NP function. In Aim 2 we’ll profile the biologic responses associated with CEP and NP exposure to matrix-modifying treatments. Using human CEP and NP explants, we’ll characterize temporal changes in matrix turnover following treatment and discover the anabolic, catabolic, and inflammatory responses of the cells. These outcomes will be related to the nature of the matrix degradation products and to the baseline degree of tissue degeneration and calcification. In Aim 3 we’ll test the relevance of CEP matrix modification by characterizing NP cell function in intact bovine discs cultured after intradiscal treatment with CEP therapies that target collagen, aggrecan, and/or mineral. Through these studies, we’ll develop a conceptual framework for CEP matrix modification that closes critical gaps in understanding regarding the effects of increasing CEP permeability on NP cell function and that establishes the functional and biologic responses to CEP treatments that act on different matrix constituents.

NIH Research Projects · FY 2026 · 2026-06

Project summary Schwann cell tumors are the most common peripheral nervous system (PNS) tumors, and people with neurofibromatosis type I (NF-1) or neurofibromatosis type II (NF-2) are at significantly increased risk to develop PNS tumors. NF-1 is caused by loss of the NF1 tumor suppressor gene while NF-2 is caused by loss of the NF2 tumor suppressor gene, but a functional link between these two neurofibromatosis tumors required for Schwann cell tumorigenesis has not been well described. Our work recently showed that NF2 is mutated in NF1 mutant malignant peripheral nerve sheath tumors (MPNSTs), an aggressive PNS tumor that is the most common cause of death in adults with NF-1. However, we do not understand if NF2 is required or sufficient for NF1 PNS tumor malignant transformation or the key signaling pathways engaged by NF2 in NF1 mutant tumors, constituting a key knowledge gap that impedes the development of better therapies for people with neurofibromatosis associated tumors. This proposal focuses on investigating if NF2 cooperates with NF1 in the Schwann cell lineage in vertebrate model organisms or tumor systems and defining whether Hippo signaling, which is dysregulated upon NF2 loss in NF1 mutant tumors, is a viable therapeutic approach for people with MPNSTs. To do so, our proposal incorporates novel mouse and zebrafish models with recently developed pharmacologic inhibitors of Hippo dysregulation and human patient MPNST samples to test the hypothesis that NF2 loss promotes NF1 PNS tumor malignant transformation through Hippo signaling. We will test this hypothesis by first determining whether Nf1/Nf2 conditional mutant mice develop MPNSTs, as Nf1 or Nf2 conditional loss alone is not sufficient for MPNST formation in mice. For added rigor and validation in an orthogonal vertebrate neurofibromatosis model, we will further evaluate whether nf1/nf2 mutant zebrafish develop MPNSTs. To complement these loss of function experiments, we will then perform inducible Nf2 overexpression in MPNST tumor models to determine whether NF2 reconstitution blocks MPNST growth in vivo. Finally, we will test the therapeutic efficacy of targeting Hippo pathway dysregulation across a range of MPNST models and aim to develop a clinical biomarker of Hippo pathway dysregulation in MPNSTs using a multi-institutional cohort with the goal of identifying a signature to predict which patients might benefit from targeting Hippo signaling. We have assembled a multidisciplinary team of biologists, oncologists, and pathologists to carry out this work comprising multiple model systems and technical approaches. The long-term goal of this research is to identify new treatment strategies for MPNSTs, which currently lack any effective therapeutic interventions, and our work will potentially establish the preclinical rationale for future clinical trials targeting Hippo signaling in these tumors.

NIH Research Projects · FY 2026 · 2026-06

Abstract A botulinum antitoxin that is renewable, reproducible, and safe, and that is suitable for rapid mass administration and facile stockpiling, is needed to replace equine BAT. We have demonstrated that highly potent monoclonal antibody combinations (mAbs) formulated with < 30 mg of total protein in under 1 mL volume can provide broad- spectrum neutralization of BoNT serotypes A-G, i.e. heptavalent neutralization. The mAb combinations were engineered to bind all known subtypes within a given BoNT serotype. The mAbs for serotypes A, B, C, D, and E have been shown to be safe in Phase I and Phase II trials with no unexpected adverse events. Antitoxin antibodies prevent the progression of disease, which is clinically significant, and clearance of BoNT from the bloodstream would likely be required along with any drug that reverses intoxication to prevent reintoxication. Evidence for the value of antibodies in treating botulism comes from a prospective randomized comparison of human botulinum immune globulin to non-immune globulin. In this study, infants treated with human botulinum immune globulin had their ICU stays reduced by 2 weeks and their hospital stay reduced by 3 weeks compared to treatment with non-immune globulin. We have engineered dual variable domain (DVD-IgG1) biepitopic antibodies using the same binding domains that have been tested in Phase I and II clinical studies. Based on our findings that three IgGs are required to provide rapid clearance, we designed DVDs that bind either two different serotypes or two epitopes of the same serotype. These are easily expressed and purified and have demonstrated potency in the mouse neutralization assays (MNA) comparable to the parental three IgG1 mixture. The use of DVDs reduces the number of antibodies required for heptavalent coverage from 16 to 8. R61 aims are (1). Optimize and characterize three DVDs for neutralization of BoNT/A and B. Starting with parental IgG1 sequences identified in NIAID-funded clinical studies, we will identify the optimal DVDs that are highly potent in vivo. (2) Design and characterize 5 DVDs binding BoNT C/D/E/F/G. The BoNT HA serotype can be included with no additional molecules. R33 Aims are: (3) Develop small-scale expression systems for the DVDs. Optimize linkers, pairings, and the order of inner and outer variable domain. (4) Co-formulate 8 DVDs as a lyophilized powder that can be reconstituted to cover 7 serotypes of BoNT. This will require optimization of our domain-specific assay to quantify each DVD in the mixture and minimize cross-reactivity. At the successful conclusion of the R33, we will have 8 DVDs ready for cell line development and IND-enabling studies. Completion of clinical development of a fully recombinant heptavalent BoNT antitoxin consisting of 8 DVDs as a lyophilized formulation that can be used to prevent and treat botulism caused by all known serotypes of BoNT and will be suitable for stockpiling at ambient temperature. The final product will be delivered intramuscularly, requiring < 30mg of total DVD protein.

NIH Research Projects · FY 2026 · 2026-06

Project Summary Our ability to manipulate primary immune cells has advanced treatments for many human malignancies. While extensive work has focused on developing genetic methods to engineer immune cells, chemical approaches remain underexplored. Chemically modifying disease-associated proteins or immune receptors offers a simple, modular, and cost-effective way to redirect cytotoxicity toward diseased cells. Such approach, however, requires organic transformations that can be performed in a crude biological milieu or in living organisms with precise molecular control. A few reactions meet these criteria, even fewer have been employed to transform primary immune cells. Additionally, the nature of the chemical modifications and geometry of the corresponding receptor-small molecule fusion are critical for facilitating optimal immune response. Therefore, developing bioconjugation strategy capable of modifying endogenous cell surface receptors, and introducing new functions, will not only enhance our ability to synthesize new biomolecules but also produce cell-based modalities for therapeutic applications. Herein, the proposed work introduces antibody-driven proximity bioconjugation to equip immune receptors with the ability to redirect and activate effector cells to specific targets. First, a panel of antibody conjugates, consisting of an electrophilic linker and a payload of choice, will be synthesized. The preparation of these reagents emphasizes modularity and balance between conjugate reactivity and stability. Mass spectrometry- based pipelines will be used to comprehensively analyze the conjugates’ ability to modify the target antigen in vitro and on cells. Reaction parameters, such as specificity and efficiency, will be correlated with the warhead’s reactivity and the antibody’s affinity, culminating into a roadmap for further optimization. Second, antibody- directed ligation will be employed to introduce small molecule adapters onto CCL-1, a key surface target of acute myeloid leukemia. This modification will enable molecular control over complementary anti-adapter CAR T cell activity and broadening therapeutic index. Finally, the programmability and modularity of the antibody-directed ligation will allow for the modification of various immune receptors with unique chemical elements. Combinatorial targeting will create opportunities for engaging multiple cooperative cell types and eliciting synergistic immune responses. Together, the proposed studies offer a creative and multidisciplinary solution to longstanding challenges in bioconjugation and engineered cell therapies. Beyond cytotoxic signaling, future work will focus on modifying and multiplexing immune receptors with agonists or antagonists to fine-tune downstream signaling. Additionally, extending antibody-directed ligation to transform other cell surface molecules will further catalyze the development of new therapeutics.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY/ABSTRACT Dr. Nielsen’s long-term goal is to understand molecular mechanisms that drive autoimmune diseases to improve their diagnosis and treatment. This five-year research and training plan will launch her independent career studying thymic T cell tolerance. To select a functional, self-tolerant T cell repertoire, developing thymocytes test their vast, random array of T cell receptors (TCRs) at key checkpoints to 1) ensure MHC restriction, 2) “tune” their capacity to distinguish self from foreign stimuli, and 3) “prune” self-reactive TCRs via clonal deletion or diversion to a regulatory T cell (Treg) fate. Nr4a transcription factors – whose expression scales with self-reactivity – collectively translate the TCR signals that enforce clonal deletion, Treg diversion, and aspects of TCR “tuning.” However, distinct protein expression dynamics for family members NR4A1 and NR4A3 suggest a “division of labor” in thymic selection which has yet to be unmasked. This proposal’s central hypotheses are: 1) NR4A1’s low threshold for induction and rapid decay allow it sense and tune TCR signaling during positive selection, 2) collective NR4A “dosage” determines deletion and Treg diversion, and NR4A3’s high threshold for induction and sustained expression enforce death and diversion of select self-reactive clonotypes in the medulla. Using biochemical assays of TCR signaling, Aim 1 characterizes how TCR tuning and NR4As operate in human thymocytes and – using genetic mouse models – determines the roles of NR4A1 and NR4A3 in TCR tuning. Aim 2 leverages high-resolution TCR sequencing and unbiased auto-antibody profiling (PhIP-seq) in to identify how the NR4As uniquely and cooperatively shape the thymic TCR repertoire. Aim 3 employs single-cell transcriptomics (CITE-seq) combined with a TCR signaling reporter to decode the transcriptional networks by which the NR4As link TCR signaling to tolerogenic outcomes during thymic selection. Completion of this project will provide unprecedent molecular insights into TCR signaling mechanisms and the formation of a tolerant TCR repertoire, illuminating fundamental principles of autoimmunity and informing strategies for T cell-based immunotherapies. Dr. Nielsen will perform this research at the University of California, San Francisco, a world-renowned institution for immunology. She has assembled a strong mentorship and collaborative team: Dr. Julie Zikherman (primary mentor, lymphocyte signaling), Dr. Mark Anderson (co-mentor, thymic tolerance), Dr. Ellen Robey (T cell selection), Dr. Mike Waterfield (gene regulation), Dr. Jimmie Ye (single-cell genomic technologies), and Dr. Wan-Lin Lo (University of Utah, TCR signaling in human cells). Through hands-on training, formal coursework, and engagement with the global immunology community, she will develop essential lab management skills and master advanced immunologic and genomic techniques, positioning herself as an independent investigator driving cutting-edge research in immune tolerance.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY The acute respiratory distress syndrome (ARDS) is associated with significant morbidity and mortality and neutrophils are critical to its pathogenesis. Neutrophil activation is regulated by inhibitory tyrosine phosphatases including the Src homology region 2 domain-containing phosphatase-1 (SHP1). SHP1 mutations are associated with severe pediatric lung disease and early onset COPD, the former is a recent novel observation by our research group. I have published data that neutrophil SHP1 prevents hemorrhage and hyperinflammation in acute lung injury (ALI) after LPS challenge or P.aeruginosa infection and pharmacologic activation of SHP1 reduces lung inflammation in the setting of ALI. SHP1-dependent intra-cellular signaling limiting hyperinflammation and pulmonary hemorrhage is unknown. I propose the central hypothesis that SHP1 mutations associated with reduced phosphatase activity will have allele-specific hyperinflammation with distinct phospho-proteomic profiles, and pharmacologic activation of SHP1 will reduce lung injury. To test this hypothesis, I will utilize recently generated novel SHP1 allelic series mice containing patient identified mutations. Aim 1 – Determine the role of SHP1 in acute lung inflammation through analysis of SHP1 allelic series mice using pre-clinical models of ALI coupled with mass spectrometry-based proteomics. Aim 2 – Test the hypothesis that pharmacologic activation of SHP1 reduces inflammation in the setting of ALI. Significance of the Results – The proposal will reveal the immunoregulatory role of SHP1 in ALI, guide the development of SHP1 activating compounds and lay the foundation for precision pharmacotherapies for patients with hyperinflammation associated lung injury. Career Development Plan – The proposed project and training plan will lay the foundation for my transition to an independent physician-scientist studying the role of immune regulating phosphatases in lung injury and inflammation. The plan involves didactics, conference attendance, mentorship, and hands-on training by experienced and highly successful primary mentor physician-scientists, in addition to expert collaborators and an experienced career advisory committee. Environment – Training will be at UCSF, which is an ideal environment that supports physician-scientists. The UCSF Department of Pediatrics has a long track record of fostering the career development of young faculty. UCSF is a highly collaborative institution and both mentors are members of the UCSF Immunology Program, which is highly regarded worldwide.

NIH Research Projects · FY 2026 · 2026-06

Project Summary: Parkinson’s Disease (PD) has motor and nonmotor features. Amongst nonmotor symptoms, cognitive impairment drives disability and loss of quality of life. Even in early PD, deficits develop in executive function, which includes cognitive flexibility. These symptoms are rarely remediated by dopamine replacement therapy, and in fact, dopamine agonists can trigger impulse control disorder (ICD). ICD manifests as compulsive gambling, shopping, pornography use, or eating. While PD-related executive dysfunction and ICD are common, little is known about their cellular or circuit-level mechanisms. In this collaborative project, we will take advantage of our labs’ (1) optimized cognitive assays in Parkinsonian mice, (2) expertise in in vivo imaging, in vivo electrophysiology, and ex vivo electrophysiology to investigate the circuit- and synaptic-level mechanisms driving PD-related executive dysfunction and ICD. In Aim 1, we will determine the role of dynorphin – a neuropeptide specifically expressed in direct pathway spiny projection neurons (dSPNs) – in behavioral flexibility in mouse models of PD. In Aim 2, we will determine the role of prefrontal cortex- dorsomedial striatum inputs in the mouse model of PD/ICD. Lastly, in Aim 3, we will use in vivo 2-photon imaging and single-unit electrophysiology to identify the neuronal ensemble activity patterns and physiological signature of dysregulated executive function. Together, this integrative project will focus on striatal plasticity mechanisms underlying the cognitive dysfunction in PD. The insights from these studies will inform the development of treatments that alleviate cognitive-behavioral dysfunction in PD and ICD. 1

NIH Research Projects · FY 2026 · 2026-06

ABSTRACT Phosphoinositide 3-kinase alpha (PI3Ka), one of the most frequently mutated proteins in cancer, converts phosphatidylinositol 4,5-biphosphate (PIP2) to PIP3, exerting widespread control over signaling pathways including Ras and Akt. In its inactive state, PI3Ka's catalytic domain, p110, is inhibited by the regulatory domain, p85, with activation involving the relief of p85-mediated inhibition by binding to phosphorylated tyrosines on receptor tyrosine kinases (RTKs) and Ras. The Ras/RTK inputs not only release the inhibition of p110 but also serve to recruit PI3Ka to the membrane and presumably promote activating interactions at the membrane bilayer. However, neither RTK nor Ras-mediated interactions with PI3Ka have been directly structurally characterized, and no structures of PI3Ka in the active state while interacting with the membrane have been reported. Given that common cancer mutations in PI3Ka, such as E542K, E545K, and H1047R, are believed to emulate these natural activation mechanisms, elucidating them holds paramount importance in clinical contexts and drug discovery endeavors. Our proposal aims to bridge this gap by characterizing PI3Ka activation at the membrane using advanced biochemical assays and cryo-EM imaging techniques established in our labs. In our preliminary studies we obtained the first cryo-EM reconstructions of the full-length PI3Ka holoenzyme in complex with Ras and phospho-peptides at the surface of lipid nanodiscs. These preliminary reconstructions reveal novel conformational rearrangements between the p110a and p85 subunits, a series of increasingly more active conformations induced by membrane binding and two novel dimeric states of PI3Ka. In Aim 1, we will use biochemical assays with PI3Ka in the presence of phosphorylated RTK tails and KRas on liposomes to understand their cooperation in PI3Ka activation and aim to obtain first high-resolution structural insights into PI3Ka complex with RTK/Ras in the context of lipid nanodiscs. In Aim 2, we will obtain high resolution structures of our newly discovered PI3Ka dimers in the context of larger membrane surface of liposomes and investigate the functional consequences of such dimers. In Aim 3, we will use a combination of mass spectrometry and signaling assays to interrogate how cooperation between RTKs and Ras as activating PI3Ka inputs is fine-tuned in cells by the specific patterns and extent of RTK phosphorylation, and PI3Ka dimerization, to generate distinct signaling inputs. Across all aims, we will investigate how cancer mutations impinge on the physiological mechanisms of PI3Ka activation unveiled by our structural and functional studies. Our ability to conduct structural studies on the full-length PI3Ka in complex with its activating inputs and membrane attachment is innovative and to our knowledge, unprecedented, and will offer a long- awaited physiologically relevant context for mechanistic investigation of oncogenic PI3Ka mutations. Since these mutations influence therapeutic sensitivity, deciphering the mechanisms by which they dysregulate PI3Ka activation could significantly alter patients' treatments.

NIH Research Projects · FY 2026 · 2026-06

Project Summary Schizophrenia (SCZ) is a severe psychiatric disorder with a complex genetic component. Genetic studies have identified a strong link between genetic risk and the neurodevelopmental processes of the brain, contributing to SCZ etiology. Notably, neurogenesis is especially vulnerable to disruption by SCZ genetic factors. However, which SCZ-associated risk genes and variants affect neurogenesis and how non-coding risk variants influence risk gene expression remains largely unknown. The overarching goal of this K99/R00 project is to systematically delineate the direct connection among SCZ-associated variants, risk genes, and neurogenesis by developing and applying advanced functional genomic screening platforms in cerebral organoids. During the K99 phase, I will elucidate the roles of SCZ risk genes in neurogenesis by conducting pooled high throughput CRISPR interference screens in key neurogenic cell types derived from cerebral organoids. During the K99/R00 phase, I will employ prime editing screens in cerebral organoids to assess the effects of individual SCZ-associated variants on neurogenesis. Finally, during the R00 phase, I will investigate the regulatory impact of non-coding SCZ variants on gene expression using single-cell prime editing screens in cerebral organoids. The successful completion of these aims will provide novel insights into which and how SCZ-associated genes and variants disrupt neurogenesis, advancing our understanding of the neurodevelopmental basis of SCZ and aiding in identifying novel therapeutic targets. Additionally, this research will enhance our ability to interpret the broader role of genetic factors in neurogenesis and will generate essential training data for machine learning models focused on neurogenesis, with potential implications for other neurodevelopmental disorders. During the K99 phase, I will further improve my expertise in functional genomics, organoid models, statistical analysis, machine learning, and neurobiology of SCZ, as well as other essential professional skills, including leadership, mentoring, writing, and presentation. To achieve my training and research objectives, I have assembled an exceptional and interdisciplinary team of mentors and collaborators including Dr. Yin Shen (primary mentor, functional genomics and gene regulation), Dr. Arnold Kriegstein (co-mentor, brain organoid), Dr. Katherine Pollard (co-mentor, statistics and machine learning), Dr. Hongjun Song (co-mentor, SCZ neurobiology), and Dr. Xin Jin (collaborator, complex in vivo screening methodologies). This comprehensive mentorship and collaborative research environment will foster my transition to an independent research career, with a long-term goal of elucidating the genomic mechanisms underpinning neurological disorders, ultimately enabling the identification of novel therapeutic targets for prevention and treatment.

NIH Research Projects · FY 2026 · 2026-06

Multidrug-resistant tuberculosis (MDR-TB) was estimated to occur 600,000 people in 2017. The roll-out of the GeneXpert™ test has generating a substantial increase in the demand for MDR-TB treatment. However, current MDR-TB treatment regimens take 9 months or longer to complete and have substantial toxicity. Therefore, a shorter, less toxic treatment regimen is needed. We have designed a regimen that does not contain PZA or an injectable agent and limits the administration of linezolid to the initial 8 weeks of treatment, before the neuropathic side effects of linezolid occur. Animal studies support the likely efficacy of this regimen. Studies of fixed duration regimens to achieve treatment shortening are associated with high risk, since there are no validated ways to predict what duration of treatment will be optimal. We have developed an innovative Phase 2 study design (“duration-randomization”) to identify the shortest effective treatment duration. In this design, participants are randomized to four durations of treatment from the shortest to the longest likely effective duration. The results are then analyzed together to determine the optimal treatment duration. In the proposed multicenter, randomized, partially blinded, four-arm, phase 2 DRAMATIC Trial (Duration Randomized Anti-MDR-TB And Tailored Intervention Clinical Trial) we will examine the efficacy and safety of an all-oral regimen of bedaquiline, delamanid, levofloxacin, linezolid, and clofazimine given for 16, 24, 32 or 40 weeks. By modeling the results of the four durations together, the design achieves substantial statistical efficiency. The optimal treatment duration identified in this trial can then be validated in a larger prospective Phase 3 clinical trial. In addition, recent studies have demonstrated that baseline patient characteristics can predict TB treatment outcomes; we will therefore stratify participants into those with “extensive” and those with “non-extensive” disease to provide guidance for clinical treatment. Aim 1: To identify the optimal duration of an experimental MDR-TB treatment regimen consistent with a successful treatment outcome. Aim 2: To describe the relationship between baseline prognostic risk strata and successful MDR-TB treatment outcome. Aim 3: To establish that the rRNA synthesis ratio, a novel biologic marker based on M. tb precursor rRNA, is associated with relapse at the individual-level across the range of durations studied in the trial. Development of a shorter, better-tolerated treatment regimen will greatly enhance the ability of TB control programs to treat the growing number of patients. The DRAMATIC Trial will employ an innovative and efficient new design to establish a robust, nontoxic MDR-TB treatment regimen and identify the minimal duration for which it needs to be administered. These results will speed the process of moving forward to a confirmatory phase 3 clinical trial and increase the likelihood that such a trial is successful. RELEVANCE (See instructions): In the proposed multicenter, randomized, partially blinded, four-arm, phase 2 DRAMATIC Trial we will examine the efficacy and safety of an all-oral regimen of bedaquiline, delamanid, levofloxacin, linezolid, and clofazimine given for 16, 24, 32 or 40 weeks. By modeling the results of the four durations together, the design achieves substantial statistical efficiency. The goals of the study are to demonstrate the safety of the regimen, to identify the shortest effective treatment duration, and to examine whether baseline participant characteristics can be used to tailor treatment duration. )RUP 3DJH FRQWLQXDWHG 3 Program Director/Principal Investigator (Last, First, Middle): Nahid, Payam

NIH Research Projects · FY 2026 · 2026-06

Summary Many cancers are treated with immunotherapy, sometimes leading to durable remissions and, in rare cases, cures. However, individual responses are variable, and few biomarkers are clinically validated to predict response. Current methods to detect on-treatment response are also not ideal. Evaluation of therapeutic efficacy, including response evaluation criteria in solid tumors (RECIST) and immune-response related criteria (irRC), is based on computed tomography (CT) and magnetic resonance imaging (MRI). However, these criteria often prove insufficient in the immunotherapy setting due to pseudoprogression caused by immune infiltration into the tumor, resulting in apparent tumor enlargement despite death of cancer cells. Another problem is the long period between initiation of therapy and assessment (typically at least 3 months). Detecting resistance earlier is critical because patients at advanced stages of cancer may have only months to live and toxicity can be severe. If resistance to a toxic and ineffective therapy could be detected quickly, the patient could transition to an alternate therapy. For many reasons, the emerging technology of hyperpolarized (HP) 13C MRI may be particularly suited to overcoming current deficiencies in monitoring immunotherapy. HP 13C MRI is a safe, quantitative and nonradioactive approach that enables real-time monitoring of metabolism. Hyperpolarization techniques enrich the 13C signal-to-noise ratio >104-fold, making metabolic imaging of molecules by MRI feasible. Clinical studies have shown that a rapid (<2 minutes) HP 13C MRI measurement of conversion of infused HP [1-13C]pyruvate to [1-13C]lactate, a measure of glycolysis, can provide information about tumor aggressiveness and detect response to therapy since glycolysis often decreases when cells stop growing or die. Recently, [1-13C]pyruvate co-polarized with 13C-urea received investigational new drug (IND) approval from the Food and Drug Administration (FDA) for studies in humans. Unlike HP [1-13C]pyruvate, HP 13C-urea is a metabolically inactive, predominantly extracellular agent whose MR signal is not affected by metabolic conversions but instead reflects blood flow, tissue perfusion, and probe distribution volume. This dual HP-pyruvate/urea agent uniquely permits simultaneous evaluation of metabolism and perfusion that can reveal the enhanced metabolism and reduced perfusion associated with poor response to systemic therapy and early relapse or disease progression of cancer. Another HP agent, [1,4-13C2]fumarate, offers the opportunity to detect cell death. Few preclinical or clinical studies have investigated immunotherapy and HP 13C MRI. The proposed project will use syngeneic mice bearing matched bladder cancers sensitive and resistant to immunotherapy to test whether changes in glycolysis, perfusion and/or cell death as measured by HP [1-13C]pyruvate/13C-urea and [1,4-13C2]fumarate MRI can detect response to immunotherapy earlier and more accurately than current standard of care imaging modalities.

NIH Research Projects · FY 2026 · 2026-06

ABSTRACT Mechanisms of parturition induction in humans remain mysterious: not only are the cellular and molecular events undertaken by the term gestation uterus prior to labor entry still largely undefined, but it is unknown whether these events are in turn initiated by some sort of “timer” that dictates overall gestation length. This fundamental knowledge gap has profound negative implications for human health since it precludes attempts to develop predictive diagnostics and rational therapies for preterm birth (PTB), a major cause of neonatal morbidity and mortality affecting ~10% of all pregnancies in the U.S. Recently, we uncovered evidence that labor onset in mice is controlled by a pathway active in uterine fibroblasts that has characteristics of a timer. The timer is “programmed” in very early gestation through genome-wide but locus-specific adjustments in levels of the repressive histone mark H3K27me3, with many loci experiencing peak accrual. Many of the same loci then experience H3K27me3 loss, a progressive process that starts soon after implantation and is associated with the induction of linked genes starting as early as midgestation. Capitalizing on these findings, this proposal seeks to identify more terminal components of the parturition cascade by determining the intra- and intercellular pathways that are controlled by such locus-specific H3K27me3 erosion, including the ones that modulate the susceptibility of mice to inflammation-induced PTB. It addresses both natural parturition as well as the pathways that operate through activation of innate type 2 immunity, which, by virtue of being hormone independent, might be more relevant to mechanisms of labor onset in humans. Specifically, Aim 1 seeks to identify candidate timer-controlled pathways of labor onset by more comprehensively mapping how the uterine fibroblast epigenome evolves over the course of gestation. This will involve assaying uterine fibroblasts isolated at successive points in gestation by ATAC-Seq and CUT&RUN for activating histone marks, combined with the use of computational tools to identify candidate target genes and gene regulatory networks that are controlled by H3K27me3 erosion. Aim 2 will then evaluate our top two current candidates, namely 1) the increase in matrix deposition, remodeling and stiffness that occurs at midgestation, a pathway that we hypothesize is driven by the timed induction of the TGF- superfamily member activin B; and 2) the timed induction of the EGF family member neuregulin-1. Finally, Aim 3 will determine how mechanisms of inflammation (LPS)-induced PTB intersect with the timer-controlled pathways that mediate how uterine fibroblasts communicate with uterine macrophages. Overall, we expect these studies to provide fundamental insight into the mechanisms of parturition onset – a problem whose solution has remained elusive across virtually all mammalian species. We also expect them to suggest both inflammatory and non-inflammatory candidate pathways that could be evaluated as potential contributors to the human labor cascade and that might be dysregulated in PTB.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY/ABSTRACT Cytomegalovirus anterior uveitis is an increasingly recognized but underestimated cause of ocular inflammation, accounting for up to 25% of anterior uveitis cases at specialized centers. While CMV infection was historically associated with retinitis in immunocompromised patients, its role in anterior uveitis among immunocompetent individuals represents a significant public health challenge that can lead to blindness through complications including glaucoma and corneal decompensation. The disease has been particularly noted in Asian and Asian-American populations, though increasing diagnostic capabilities suggest it affects all demographic groups. Recent evidence suggests multiple anterior chamber paracenteses may be required for accurate diagnosis, indicating current prevalence estimates likely underestimate the true disease burden. Treatment strategies may include oral antiviral therapy with valganciclovir but can be expensive and requires laboratory monitoring for side effects of renal toxicity and bone marrow suppression. Topical antiviral therapy with ganciclovir eye drops is an alternative approach but requires medication compounding and expires quickly. Yet another treatment approach may be simple observation without using any antivirals. Despite these options, the optimal therapeutic approach remains undefined due to a lack of comparative trials. Furthermore, while long-term antiviral suppression may prevent disease recurrence, no randomized controlled trials have established efficacy or investigated how mutations might lead to treatment resistance. Here we seek to enroll participants with PCR-proven CMV anterior uveitis into two double-masked placebo- controlled randomized clinical trials in the US and Asia to 1) compare CMV viral load and inflammation after randomization to 7 days of oral valganciclovir, topical ganciclovir 2%, or placebo (Trial I); 2) evaluate the effect of long-term antiviral suppression on CMV recurrence after achieving a clinical quiescence (Trial II); and 3) investigate viral resistance patterns and host immune responses using advanced genomic analysis of aqueous samples from both trials to understand mechanisms of recurrent disease. This proposal will establish evidence-based protocols for both acute treatment and long-term management of CMV anterior uveitis. Our innovative approach combines microbiologic and clinical outcomes with molecular analysis to understand disease mechanisms, potentially identifying biomarkers for recurrence and antiviral resistance. These findings will directly impact patient care by optimizing treatment approaches and preventing vision loss in this significant subset of uveitis patients.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY Memory is an essential cognitive process dependent on the consolidation of experience into stored memories and generalized knowledge. We know not all memories are stored, and we know memories undergo a transformation from episodic events into abstract understanding. However, the neural underpinnings of this process of memory consolidation remain unclear. The hippocampus is a brain region critical for memory formation, and the medial prefrontal cortex (mPFC) is a brain region involved in memory storage and abstracted knowledge; these regions are bidirectionally connected and are candidate brain networks for the selection of memories for consolidation and the transformation of memory traces into generalized knowledge. Further, the sleep sharp-wave ripple (SWR) is a brain oscillation known to be involved in memory consolidation during which privileged hippocampal-mPFC communication occurs. While systems consolidation theory offers predictions of how the hippocampus and mPFC interact during SWRs to consolidate memories, these predictions have so far been difficult to causally test in the absence of multi-site neural recordings with optogenetic manipulations. To test the hypothesis that cortical-hippocampal information flow preceding SWRs is critical for the selection of memories for consolidation, this project aims to silence mPFC activity in conjunction with simultaneous large- scale electrophysiology recording of the hippocampus. This will determine the role of the mPFC in influencing hippocampal activity during SWRs, providing a mechanism by which memory traces are selected for consolidation (Aim 1). In addition, to test the hypothesis that hippocampal-cortical information flow during SWRs is critical for the emergence of consolidated, generalized cortical representations, I will specifically inhibit mPFC activity during SWRs. This will evaluate whether mPFC activity during this brain oscillation is necessary for the transformation of memory traces and the development of neural representations of generalized knowledge (Aim 2). Completion of these aims has the potential to yield fundamental insights into the neural mechanisms of memory consolidation. This study will be carried out in the lab of research sponsor, Dr. Loren Frank, at the University of California, San Francisco (UCSF). The Frank Lab is located in the Sandler Neurosciences Center, which is home to a highly innovative and collaborative community of faculty and students pursuing neuroscience investigation. Pursuing this project will accomplish the training goals of gaining expertise in in vivo electrophysiology data acquisition, developing quantitative data analysis skills, and improving my scientific communication. The training plan under this fellowship will provide preparation for an independent career as an academic neuroscientist-neurologist with the long-term goal of revealing neural circuits underlying cognitive processes and flexible behavior. In addition to the proposed research, this preparation will be achieved via composition of scientific manuscripts, engagement with vibrant intellectual communities, and neurology-geared clinical activities.

NIH Research Projects · FY 2026 · 2026-06

ABSTRACT This proposal presents an innovative and interdisciplinary approach to accelerate the development of effective, precision therapeutics for Alzheimer’s disease (AD), a complex and heterogeneous neurodegenerative disorder with limited treatment options. Recognizing the critical role of neuroimmune and vascular dysfunction in AD pathogenesis—particularly within the perivascular spaces, the we will leverage state-of-the-art single-nucleus transcriptomic profiling (VINE-seq) and integrative computational drug repositioning to identify and test repurposed compounds capable of modulating disease-relevant gene expression signatures at the cell type level. The project builds on extensive preliminary data identifying key AD-related transcriptional signatures across neurons, glia, and vascular/perivascular cells, revealing both known and novel therapeutic targets. Using publicly available and lab-generated transcriptomic datasets, the team employs tools like Connectivity Map and LINCS to predict drug candidates that reverse pathogenic gene networks in distinct brain cell types. Early analyses have already nominated over 80 potential compounds, including letrozole, irinotecan, sirolimus, and vorinostat, several of which show multi-compartment activity and strong preclinical promise. These candidates will be rigorously validated in vitro using iPSC-derived neurons, glia, and assembloid models, and in vivo using mouse models of AD with tools like longitudinal bioluminescent imaging and plasma biomarkers to assess efficacy, target engagement, and safety. Aim 1 seeks to validate and optimize repurposed drugs targeting neuron-glial dysfunction by integrating computational predictions with functional studies in co-culture systems, chronic mouse dosing models (e.g., 5xFAD, hTAU), and multi-omic readouts of parenchymal pathology, including amyloid burden, neuroinflammation, and synaptic loss. Aim 2 focuses on the vascular-perivascular axis of AD by using single-cell signatures from VINE-seq to identify and test compounds that reverse vascular dysfunction and immune dysregulation in CAA-associated AD models (e.g., TgAPP23), with endpoints including cerebral amyloid angiopathy, blood-brain barrier integrity, and perivascular inflammation. Together, these complementary aims form a robust platform to evaluate and prioritize sex- and cell-type-specific therapies that address both neuronal and vascular drivers of AD. The collaboration brings together a world-class team with expertise in AD genomics, drug repurposing, systems immunology, mouse modeling, and human cell-based models. By uniting cutting-edge transcriptomic technologies with in vivo pharmacology and a precision medicine framework, this work has the potential to deliver impactful therapeutic strategies that can be rapidly translated into clinical trials for patients with Alzheimer’s disease.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY Self-reactive B cells must distinguish self from foreign antigens (Ag), mounting immunogenic responses to the latter but not the former. However, the mechanisms they employ to do so are not fully understood. Long-term, we aim to exploit new molecular insight gained by filling this knowledge gap to restore B cell tolerance in autoimmunity, enhance anti-tumor B cell responses to self-like Ag, and optimize host defense. In this proposal, we leverage a unique and modular platform of liposomes decorated with model Ag, enabling independent control of epitope affinity and density. Using this system, we recently demonstrated that such particulate Ag are much more potent activators of B cells than identical Ag in soluble, monovalent form. We found that this stems not merely from avidity but from the ability of particulate Ag to evade inhibitory signaling pathways normally engaged by soluble Ag and mediated by the Src family tyrosine kinase Lyn. Moreover, we discovered that transcriptional programs and B cell functional responses to particulate and soluble Ag diverge markedly; despite an identical epitope recognized by the BCR, we found that particulate Ag produce robust NF- kB activation even in the absence of T cell help, while soluble Ag drive an NFAT-associated anergy program. We propose that biophysical characteristics of particulate Ag display serves as a stand-alone danger signal that evades tolerogenic transcriptional programs and elicits immunogenic responses by B cells. In this grant, we propose to: (1) Identify proximal biochemical pathways downstream of the Lyn Src family tyrosine kinase that enable B cells to distinguish tolerogenic and immunogenic Ag display. We will edit mouse and human B cells in vitro and in vivo to test candidate substrates of Lyn. We will complement this with an unbiased proteomics approach to identify novel negative regulators of B cell responses to self-like soluble Ag. (2) Elucidate how early signaling events triggered by particulate Ag are transformed into immunogenic transcriptional programming of B cell fate through robust activation of NF-kB but not NFAT pathways. We will take a genetic approach to manipulate the second messenger diacylglycerol (DAG) which we hypothesize toggles between these immunogenic and tolerogenic B cell transcriptional responses. We will test this in human B cells. We will pursue a genomic strategy to survey the epigenetic landscape of B cells in order to define the transcriptional architecture that translates pattern of Ag display into B cell fate. (3) Systematically map the lower boundary of Ag affinity, density, and particle dose, and test the biophysical properties (particle size, membrane fluidity, linker length) required to activate B cells, including anergic self- reactive cells. We will test the hypothesis that these thresholds are controlled by Lyn-dependent inhibitory signaling pathways.

NIH Research Projects · FY 2026 · 2026-06

Project Summary/Abstract The pituitary gland plays a central role in various physiological functions, including growth, lactation, stress response, reproduction, metabolism, and water balance. The pituitary lineage-specific transcription factor PIT1 specifies three pituitary lineages that produce growth hormone (GH), prolactin (PRL), and thyroid-stimulating hormone (TSH). Naturally occurring mutations in PIT1 can cause combined pituitary hormone deficiency (CPHD) in humans, leading to deficiencies in these hormones. We previously found that PIT1 associates with a cell-type- specific chromatin organizer, SATB1, and β-catenin. This association is crucial for tethering PIT1-bound enhancers to an insoluble, salt-extraction-resistant subnuclear structure, and this tethering is consequently required for PIT1-mediated gene expression. The R271W mutation in PIT1 associated with CPHD causes failure of PIT1 to associate with SATB1 and connect with the nuclear substructure to produce GH. By developing an alternative ChIP-seq protocol (ureaChIP-seq), we recently reported that in vivo, SATB1 directly binds to specialized genomic regions called base-unpairing regions (BURs, ~300 base pairs in length) genome-wide in both mouse and human genomes. By comparing SATB1 direct versus indirect chromatin association, we found that SATB1 forms a “two-tiered” type of chromatin organization, consisting of (a) SATB1- bound BURs within the insoluble subnuclear structure and (b) accessible chromatin that SATB1 indirectly interacts with, presumably through its association with other nuclear factors bound to regulatory regions. This two-tiered organization is distinct from chromatin organized into topologically associating domains (TADs) by CTCF and cohesin. Based on our findings, we hypothesize that the three-dimensional chromatin organization regulated by SATB1 enables PIT1 to establish a cell-type-specific transcriptional program in the pituitary gland. To test this hypothesis, in Aim 1, we will use a GH-producing GC cell line to determine whether BURs targeted by SATB1 provide the chromatin scaffold to support the PIT1-regulated gene regulatory network that drives proper gene expression. In Aim 2, we will employ a novel and modified auxin-inducible degron (AID2) system to reversibly deplete SATB1 protein in vivo in a cell-type-specific manner. Using the AID2 system, we will investigate SATB1’s role in postnatal pituitary gland expansion, development, and the function of PIT1+ cells in mice. We will explore whether the SATB1-mediated two-tiered chromatin organization facilitates PIT1 association with its target enhancers in vivo. We envision that the results from this proposed research, linking chromatin architecture to cell-type-specific gene expression, will significantly advance our understanding of how the pituitary gland grows, matures, and maintains its function during adulthood. The proposed studies on the novel function of SATB1 in the pituitary gland are likely to guide the development of future therapies for pituitary- related disorders.

NIH Research Projects · FY 2026 · 2026-06

Project Summary/Abstract Targeted radionuclide therapy is resurging as an attractive avenue for cancer treatment, which can potentially be applied to various cancers independent of pathology. However, existing radiopharmaceuticals struggle with balancing safety and efficacy. Radiopharmaceuticals with high molecular weights are effective in tumor uptake yet meanwhile increase radiation exposure of normal tissues, whereas small molecule radiopharmaceuticals reduce undesired systemic radiation but also decrease tumoral absorption. While small molecule radioligands irreversibly bound to target have been shown to improve in vivo imaging, no protein-based radiopharmaceutical has been enabled to bind target irreversibly. To change this paradigm, this project seeks to develop a new class of covalent protein radiopharmaceuticals. New latent bioreactive amino acids will be designed and genetically incorporated into the protein binder of low molecular weight, followed with efficient radionuclide labeling. When administrated in vivo, the low molecular weight of the protein will allow rapid clearance of the radiopharmaceutical from circulation, ensuring low background and safety; Upon recognition of the target by the protein binder, the latent bioreactive amino acid in the binder will react with a natural residue in the target, cross-linking the radiopharmaceutical to the target irreversibly. This covalent and irreversible binding will prolong the radiopharmaceutical’s residence time at the tumoral site and enhance absorbed dose. The pharmacokinetics and biodistribution of the covalent protein radiopharmaceuticals will be assessed in tumor bearing mouse models of human cancer using positron emission tomography, and their antitumor effects will be evaluated in cancer cell xenograft mouse models and clinically relevant patient-derived xenograft mouse models. Mouse studies are critical as only small animal models can recapitulate whole-body radiotracer biodistribution and enable human dosimetry prediction - capabilities that cannot be achieved using cell lines, organoids, or similar in vitro systems. The success of this project will unite an unusual combination of safety with efficacy for protein radiopharmaceuticals, enabling a new class of protein radiopharmaceuticals with expanded therapeutic index for cancer treatment.

NIH Research Projects · FY 2026 · 2026-06

Project Summary/Abstract Background: The proposed project is aligned with NIH’s Strategic Plan for Nutrition Research (SPNR) Obj. 4- 2&3, to reduce the burden of malnutrition in clinical settings. Up to 40% of patients with anorexia nervosa (AN) become medically unstable due to malnutrition and require hospitalization for refeeding; 25% progress to severe and enduring illness. Long, intensive hospitalizations with frequent readmissions drive high healthcare costs in AN. Historically, clinicians have relied on body weight to assess malnutrition and response to intervention. However, body weight lost diagnostic power due to rising BMIs. About 1/3 of our patients today are diagnosed with atypical AN (AAN)—with medical instability at “normal” weight. The upward shift in BMI is reflected globally, leading new recommendations to include body composition to diagnose malnutrition. Emphasis is on low fat free mass (FFM) as a predictor of poor hospital outcomes. However, this has not been examined in hospitalized patients with AN, who are often too medically unstable to transport for research scans. We can now fill this gap with whole-body, infrared, 3-dimensional optical imaging (3DO) at the bedside. We showed excellent concordance between 3DO and dual X-ray absorptiometry (DXA) for detecting malnutrition at low BMI and captured changes in FFM with bedside 3DO during refeeding. Proposed project: We will employ a functional approach to body composition, integrating compartment mass with physiologic function, to assess malnutrition and predict short- and long-term refeeding outcomes across the malnutrition continuum. Prior research on FFM has focused on the skeletal muscle and bone components and long-term risks in AN. In contrast, organ residual mass (ORM)—which comprises 43% of FFM—has received little attention despite its central role in refeeding. Profound ORM depletion in AN (loss of 43% cardiac, 22% renal, and 39% hepatic mass) contributes to organ dysfunction, hypometabolism, and refeeding complications. We will generate ORM reference values from large, representative datasets, calculate ORM index z-scores (ORMIz), and examine their association with malnutrition at baseline, in response to short-term refeeding intensity and as a predictor of long-term outcomes in patients across the continuum of malnutrition due to AN. Purpose, hypotheses and design: This multicenter, prospective, observational study will include N=90 hospitalized 12-26 yr olds with medical instability and malnutrition due to AAN, AN or extreme AN. Aim 1) Assess clinical utility of ORMIz at admission. Lower baseline ORMIz will: H1) correlate with malnutrition markers, H2) associate with pre-admission energy imbalance, and H3-Primary) predict refeeding intensity. Aim 2) Monitor response to refeeding in hospital. Change in ORM will associate with: H1) medical stability, H2) metabolic stability, and H3) lower baseline FM. Aim 3) Predict long-term outcomes. Lower discharge ORMIz will predict poor outcomes at 3, 6, 9, and 12 months. Findings will be rapidly translated into individualized refeeding approaches.

NIH Research Projects · FY 2026 · 2026-06

Project Summary/Abstract . Dr. Taylor LaFlam is a pediatric rheumatologist and early career basic science researcher. The overarching goal of this proposal is to advance understanding of B cell tolerance and trafficking while providing the scientific and career development needed to enable Dr. LaFlam to become an independently funded laboratory investigator. The clinical impetus for this research is that systemic lupus erythematosus causes significant morbidity and remains difficult to treat. B cells are clear contributors to its pathogenesis, but the mechanisms by which B cell tolerance is disrupted in lupus remain poorly defined. Dr. LaFlam’s primary mentor, Dr. Jason Cyster, in collaboration with the Vinuesa group, previously found an association between lupus and P2RY8, a G-protein-coupled receptor. The Cyster lab has identified the ligand of P2RY8 and elucidated its role in regulating migration and proliferation in germinal center B cells. Prior work has also suggested a role for P2RY8 in B cell negative selection and plasma cell generation. Under the mentorship of Dr. Cyster and Dr. Chun Jimmie Ye, a genomicist and immunologist, Dr. LaFlam has taken a functional genomics approach to P2RY8. He has advanced understanding of the molecular function of P2RY8 by near comprehensive profiling of missense variants, delineating their effect on expression and function, while also coordinating collaborating work that determined the structure of the protein. He has shepherded the creation of a new human P2RY8-expressing mouse model. Building on his prior work, Dr. LaFlam will interrogate the hypothesis that P2RY8 promotes peripheral B cell tolerance and restrains plasma cell generation and bone marrow homing. The specific aims of the proposed research are: (1) to define the role of P2RY8 in transitional B cell negative selection, (2) to delineate the role of P2RY8 in plasma cell generation and trafficking, and (3) to decipher the initiation of the signal from P2RY8 that restrains proliferation. Through this grant, Dr. LaFlam will receive training in key scientific methodologies as well as professional development that will equip him to fund and lead a successful immunology research lab. Mentorship is a critical component of this training, and in addition to Dr. Cyster as primary mentor and Dr. Ye as co-mentor, he will be advised by Dr. Julie Zikherman, a rheumatologist-scientist who studies B cell responses, and Dr. Michael Waterfield, a pediatric rheumatologist-scientist who studies mechanisms of immune tolerance. Together, the research accomplished and the skills gained through this award will enable Dr. LaFlam to become an independent, NIH-funded investigator whose work on B cell tolerance and lupus pathogenesis opens avenues to improved treatments for autoimmunity.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY Spinal cord injury (SCI) produces a devastating syndrome that is characterized by motor and sensory dysfunction. In the human clinical population, SCI is often accompanied by concomitant injuries that vary from bone fracture, skin abrasion, and lacerations (polytraumatic SCI). These peripheral injuries likely contribute to spinal cord dysfunction, but their impact is largely unknown. The impact of peripheral injury on the spinal cord below the SCI may be overlooked. We have previously used several models of peripheral injury to demonstrate that polytraumatic SCI exacerbates neuroinflammation and maladaptive spinal plasticity, and can act to further undermine recovery of locomotor/sensory function. Interestingly, neuroinflammation contributes mechanistically to lumbar cord dysfunction below SCI. We have also demonstrated that peripheral nociceptive input increases and prolongs the inflammatory cytokine expression profile in the injured spinal cord. These findings suggest that polytrauma creates a vulnerability to neuroinflammation after SCI. The proposed R01 will explicitly test the hypotheses that 1) the precise immunomodulatory mechanism(s) by which peripheral injury alters spinal cord function after SCI, 2) the contribution of peripheral circulating immune cells, and 3) whether general anti-inflammatory or targeted block of peripheral immune cells reduce spinal cord neuroinflammation and improve recovery of behavioral function in polytraumatic SCI. We will test three models of peripheral injury (hindpaw capsaicin; spared nerve injury; tibial fracture) combined with contusive SCI, followed by a central anti-inflammatory (i.t. sTNFR1) or a systemic CCR2 antagonist (i.p. INCB3344) to assess the unique contributions of central neuroinflammatory vs. peripheral immune/inflammatory responses. Tibia fracture is hypothesized to cause more peripheral inflammation by mobilizing resident bone marrow-derived monocytes into circulation, while pure nociception via TRPV1 and nerve injury are hypothesized to cause central inflammation via direct tetanic afferent input into the CNS. Aim 1 will employ RNAseq with advanced gene network analyses and histopathology to demonstrate how early neuroinflammatory blockade alters the distinct pathways and injury processes engaged by different forms of polytraumatic SCI. In Aim 2 we will use single cell mass cytometry (CyTOF) to identify the unique profile of pro- inflammatory cell populations that respond to polytraumatic SCI and verified against anatomical markers of injury, to determine how these cell types (eg resident microglia vs peripheral macrophages) are affected by early block of neuroinflammation. Aim 3 will use a battery of behavioral and sensory assessments to test whether the early targeted anti-inflammatory therapy restores long-term function after polytraumatic SCI. Together these aims are designed to identify the distinct roles of peripheral and innate immune response and central neuroinflammation as drivers of persistent behavioral and sensory dysfunction in response to polytraumatic SCI, and to test the therapeutic efficacy of two anti-inflammatory precision-medicine approaches.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY/ABSTRACT Animals exhibit a remarkable array of flexible behaviors. Birds alternate between caching and retrieving food based on availability; rats reroute when familiar paths are blocked; humans revise strategies mid-game in chess. This ability to flexibly switch strategies or generate new solutions is central to intelligent behavior and is often impaired in neuropsychiatric disorders such as autism spectrum disorder and schizophrenia. Prior research has yielded key insights into what supports such cognitive flexibility: internal models of the world, including spatial and episodic knowledge encoded in the hippocampus (HPC) and abstract rules encoded in the prefrontal cortex (PFC). However, we still lack a mechanistic understanding of how the brain engages these models in real time to guide strategy switching and problem-solving. This proposal addresses this gap by identifying internal strategy states—latent variables computed by the brain that track the currently active policy for selecting goal-directed actions—and by dissecting the neural computations that encode, update, and drive transitions between these states. I will combine large-scale electrophysiology with closed-loop optogenetics in freely behaving rats performing strategy-switching and problem-solving tasks. I will assess behavioral and neural data by integrating two complementary theoretical frameworks: (i) reinforcement learning and Bayesian inference to formalize latent behavioral strategies and valuation processes; and (ii) dynamical systems modeling to uncover how neural population activity implements these cognitive operations. I will test the central hypothesis that the flexible control and generation of strategies arise from structured population dynamics in medial PFC (mPFC) implementing computations that: (i) direct HPC to simulate future scenarios that inform strategy switching (Aim 1); (ii) track and update strategy values to determine when to switch (Aim 2); and (iii) integrate input from the orbitofrontal cortex to select among multiple strategies and generate new solutions (Aim 3). By causally linking neural dynamics to strategy switching, selection, and generation, this work will reveal algorithmic and implementational principles of cognitive flexibility, laying the groundwork for my long-term goal: to elucidate the division of computational labor across PFC subregions and their interactions with subcortical regions (e.g., thalamus) during multi-strategy problem-solving. The K99 phase will support my transition to independence through training in multi-region, high-density electrophysiology coupled with real-time optogenetics, as well as advanced behavioral and dynamical systems modeling. I have assembled a mentorship team (Drs. Loren Frank, Joshua Berke, and Nathaniel Daw) and collaborators (Drs. Vikaas Sohal and Scott Linderman) with complementary expertise spanning experimental, technological, and theoretical domains of systems neuroscience. This award will also provide professional development in lab management, leadership, scientific communication, and grant writing, which will position me to launch an independent research program focused on the neural basis of intelligence and creative behavior.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY Natural killer (NK) cells are critical members of the immune system tasked with recognizing and eliminating harmful cells, including virally infected and cancerous cells. They recognize harmful cells (target cells) through membrane bound NK receptors (NKR) that interact with ligands on the surface of targets. Upon cross-linking of activating NKRs, immunoreceptor tyrosine-based activating motifs (ITAM) on adaptor molecules get phosphorylated and initiate signaling cascades that result in the activation of NK cells – and ultimately kill targets. Since their discovery, ITAMs have always been regarded as only driving activating signals. However, recent studies suggest that there are nuances to this activation and there may be scenarios where ITAMs suppress immune cell activation. Given that NK cells play a critical role in protection from harmful cells and that many of their activating NKRs depend on ITAM-containing adaptor molecules, understanding the signaling cascades that achieve optimal NK cell activation is fundamental to enhance cellular therapies. This is particularly important as numerous reports demonstrate that immune cells face a strong immunosuppressive environment in some diseases, including in the tumor microenvironment. In this proposal, we will investigate the roles that ITAMs play in activating NK cells. We will achieve this by using mouse models whereby their NK cells express mutant adaptor molecules whose ITAMs are incapable of activation. Using these mutant ITAM-deficient mice, i) we will determine if ITAMs influence NK cell development, ii) if ITAMs influence phenotypic differences in NK cells, and iii) if ITAMs calibrate the responsiveness of NK cell to harmful cells. We will also begin to elucidate if signaling through ITAM- bearing receptors impacts cellular fates of NK cells. NK cells have been documented to exhibit adaptive-like features with enhanced secondary responses and increased longevity, therefore understanding the factors that promote adaptive NK cell formation are critical. To achieve these answers, we will use in vitro and in vivo models of cancer and viral infection to assess NK cell function and use cutting-edge technologies to assess NK cell phenotypes. In summary, these studies will investigate the importance that ITAMs contribute to overall NK cell fitness and driving NK cells with enhanced protective features. These studies have the potential to greatly enhance NK cell based cellular therapies in the clinic.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY The prevalence of type 2 diabetes (T2D) is increasing in adolescents, and metformin, the first-line FDA-approved therapy for T2D, often fails to achieve durable glycemic improvement. Glucagon-like peptide-1 receptor agonists (GLP-1 RAs) such as semaglutide have transformed the treatment landscape, offering substantial benefits, including weight loss, glycemic improvement, and β-cell preservation. However, their unintended effects on skeletal muscle loss and eating behaviors in adolescents remain poorly understood. Understanding these benefits is crucial for optimizing treatment safety and efficacy during adolescence, a key developmental period. To address this unmet medical need, Sujatha Seetharaman, MD, proposes a study through this career development with the following aims (1a) assess changes in body composition and functional outcomes over six months of semaglutide therapy in adolescents with T2D (1b) examine how the rate of weight loss with semaglutide relates to changes in muscle mass in adolescents with T2D (2) investigate the effect of semaglutide on the risk of disordered eating and internalized weight stigma in adolescents with T2D. She will enroll 50 adolescents (ages 12–17.99 years) with new-onset T2D from UCSF Diabetes Clinics, treated with metformin for at least 3 months, and initiating semaglutide per their routine clinical care. In aim 1, she will assess changes in fat mass using magnetic resonance imaging, skeletal muscle mass using the D3 creatine dilution method, muscle quality, strength, and function at baseline and 6 months. In aim 2, she will calculate the rate of weight loss over 6 months and correlate it with skeletal muscle mass loss, strength, and function. In aim 3, she will examine changes in eating attitudes and internalized weight-related stigma at baseline and at 6 months to identify individuals who are at risk for disordered eating. Results from this proposal will inform an R01 in which targeted interventions such as an integrated exercise program and eating disorder prevention strategies will be tested in a large randomized controlled trial and expanded to include emerging dual/triple agents in adolescents with T2D. Dr. Seetharaman’s long-term career goal is to become an independently funded clinician-investigator advancing research that integrates novel pharmacotherapies with tailored interventions to improve health outcomes in adolescents with T2D. In this career development award, her career goals are to gain targeted training in body composition analysis, become proficient in advanced biostatistical methods, become proficient in utilizing person-reported outcome metrics, gain experience in conducting pediatric clinical trials, and engage in career development activities in preparation for R01 funding. She will accomplish these goals with the advice and mentoring from a world-class multidisciplinary mentoring team, participation in relevant didactic coursework, and hands-on research experience, all necessary for future independent success. This project aligns with NIDDK’s mission to improve the health and well-being of individuals affected by diabetes and related conditions.

NIH Research Projects · FY 2026 · 2026-05

Summary/Abstract: The multifunctional cytokine TGF-β is a central mediator of the chronic inflammatory and fibrotic pathologic processes that leads to lung and airway fibrosis. Our long-term goals for this project are to acquire a deeper understanding of how TGF-β function is regulated, and to leverage that understanding to develop new strategies and treatments for fibrosing lung disease. There is an urgent need for effective therapies to treat chronic fibrosing and inflammatory diseases of the lung. Clinical trials targeting TGF-β itself or its receptors has faced challenges, most notably, toxicities. More understanding of how TGF-β functions is clearly needed to develop better and more specific methods to more effectively target the pathologic effects of TGF-β. Since TGF-β is always produced in an inactive form that must be activation to function, methods targeting its activation may more specifically target the local effects of TGF-β thus avoiding systemic toxicities. TGF-β is activated by binding to several integrins. What exactly happens biologically after these integrins bind to TGF-β remains largely speculative. Because of this lack of biologic understanding it has long been assumed that TGF-β must be released from LAP so that free TGF-β can operate as a paracrine factor and diffuse and bind its receptors on target cells. Based on our recent structural data obtained using single particle electron cryomicroscopy (cryo-EM), we have developed a new hypothesis where integrins can bind to L-TGF-β on cells presenting the L-TGF-β on their cell surface and induce autocrine signaling without release and diffusion of TGF-β. We have recently verified that such an autocrine mechanism is sufficient to maintain the essential functions of TGF-β1 in mice, and that paracrine TGF-β1 signaling is dispensable. We now have structural and cell-based data that provides a new hypothesis for TGF-β1 activation: The inherent disorder (entropy) present in specific domains of TGF-β1 and integrin αvβ8 is redistributed upon binding of L-TGF-β1 to αvβ8 which is sufficient to cause exposure of mature TGF-β1 to its receptors. Here in three aims, we address three critical questions concerning this new model of L-TGF-β activation. (1) Can entropy redistribution of the L-TGF-β/αvβ8 complex be predicted by structural methods, and if so can it be biochemically and therapeutically manipulated? (2) How does a native membrane environment affect entropy redistribution of the L-TGF-β/αvβ8 complex? (3) Can the entropy redistribution hypothesis be applied to other integrins and can it be harnessed to be therapeutically useful to treat lung fibrosis? To answer these questions, we will establish and employ state-of- the-art techniques including cryo-EM coupled to orthogonal spectroscopic, and analytical techniques to more quantitatively assess protein thermodynamics, protein engineering, recombinant antibodies to affect local protein conformational dynamics and finally testing of those antibodies in lung fibrosis models in vivo. Together, these studies will improve mechanistic understanding of TGF-β activation and therapeutic targeting strategies to inhibit it.