University Of California, San Francisco

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY Cells organize biochemical reactions into biological condensates. P-bodies are conserved cytoplasmic condensates enriched in factors important for mRNA storage or degradation, but how these opposing outcomes may be achieved in condensates is unclear. A critical step in mRNA degradation is the removal of the 5'-7- methylguanosine cap by the decapping enzyme complex (Dcp1/Dcp2) that precedes and permits digestion of the mRNA body by conserved exoribonucleases. We have reconstituted biological condensates containing fission yeast Dcp1/Dcp2 and an enhancer of decapping protein 3 (Edc3), which are major core proteins of P- bodies. Using novel, activity-based fluorescence probes we have made two significant discoveries. First, contrary to the popular model that condensates enhance enzymatic reactions due to local concentration effects, we find that phase separation represses the activity of Dcp1/Dcp2 100-fold compared to dilute solution. Second, the decapping activity of these condensates can be rescued by Edc3. Our data suggest the protein interaction platform Dcp1 is an integrator of short-linear protein interaction motifs that couples phase separation to inactivation of decapping by promoting a conformational change in Dcp1/Dcp2 to an autoinhibited conformation. In Aim 1, we will determine the structure of the autoinhibited conformation of Dcp1/Dcp2 and test the hypothesis that short-linear motifs in Dcp2 directly interact with Dcp1 to promote a transient inactive conformation of the decapping complex. In Aim 2, we will study how condensates provide an additional layer for decapping repression, testing the hypothesis interactions that promote phase separation further stabilize the inactive conformation of Dcp1/Dcp2 in condensates. In Aim 3, we will determine the mechanism for activation of decapping in condensates, testing the hypothesis that Edc3 opposes the inhibitory action of short-linear inaction motifs in Dcp2 and promotes a conformational change that opens the RNA binding channel in Dcp1/Dcp2 to promote efficient decapping within condensates. Lesions important for repression of decapping in vitro will be tested for their function in EDC3-mediated mRNA decay in fission yeast. The proposed studies are poised to provide a paradigmatic example of how biological condensation is coupled to conformational control of enzyme activity that affects the fidelity of gene expression at the level of mRNA decay.

NIH Research Projects · FY 2026 · 2023-04

ABSTRACT Vitamin D deficiency is a widespread problem among the elderly and can precipitate sequelae that are particularly harmful to this vulnerable population. Hyperparathyroidism (HPT) is commonly associated with low vitamin D status, and the disruption in calcium homeostasis that is the cardinal feature of this disease can lead to a range of deleterious outcomes among the elderly, including loss of bone density, muscle weakness, and increased risk of falls and pathological fracture. As the primary endocrine organs responsible for hormonal control of serum calcium levels, the parathyroid glands are a known target for the actions of vitamin D, yet there are major gaps in our understanding of how vitamin D deficiency and attenuated vitamin D receptor (VDR) signaling contribute to the etiology and clinical presentation of HPT. The mechanistic intermediates linking the VDR to parathyroid hormone (PTH) hypersecretion in HPT remain unknown, and the current model of VDR- dependent expression of the calcium sensing receptor (CaSR) in the parathyroid does not account for the significant proportion of HPT cases where downregulation of CaSR abundance in parathyroid tissue is not observed. Recently, several key findings from our group suggest a testable new model for vitamin D-mediated actions in the parathyroid gland. First, we showed that the GABA B1 receptor (GABAB1R) can form heterodimeric complexes with CaSR, promoting tonic hypersecretion of PTH by opposing the coupling of CaSR with its obligate downstream G-protein effectors Gq/11 and Gi. Second, building upon a recent report demonstrating that soluble peptide derivatives of the amyloid precursor protein (APP) can bind and activate GABAB1R in neurons, we found that the APP-derived peptide Aβ1-42 can increase maximal PTH secretion by parathyroid tissue in a CASR- and GABAB1R-dependent manner. VDR expression and serum vitamin D levels are inversely correlated with the relative abundance of APP, Aβ1-42, the γ- and β-secretases required for Aβ1-42 production, and the phosphorylated form of the microtubule associated protein Tau (pTau). Functionally, ablation of APP in the parathyroid abrogates the development of HPT in VDR KO mice, and inhibitors of Tau phosphorylation can block the ability of Aβ1-42 to promote PTH hypersecretion. These data suggest that HPT driven by loss of VDR activity could arise at least in part through unregulated expression of Aβ1-42 and pTau. Based on these findings, we hypothesize that aging-induced increases in Aβ1-42-mediated signaling drive tonic PTH hypersecretion and that vitamin D deficiency exacerbates HPT disease severity by relieving suppression of Aβ1-42 production and Tau/pTau expression. To test this model, we propose three complementary, mechanistic specific aims: (1) to delineate the molecular actions of Aβ1-42 on CaSR, GABAB1R, and downstream signaling events that promote PTH hypersecretion; (2) to determine whether blocking the production or activity of Aβ1-42 and Tau delays HPT development in a murine model of CaSR insufficiency; and (3) to delineate the causal relationship between VDR and Aβ1-42/pTau signaling in the parathyroid. By defining the contributions of the CaSR/GABAB1R/Aβ1-42 signaling axis to PTH hypersecretion, this work will provide a clearer mechanistic understanding of the unexpected connection between β-amyloid peptides, vitamin D status, and parathyroid gland function.

NIH Research Projects · FY 2026 · 2023-04

PROJECT ABSTRACT After damage to white matter tracts (WMI) in CNS diseases such as multiple sclerosis (MS) in adults and newborn brain injuries that cause cerebral palsy (CP), myelin sheaths can be regenerated by activated oligodendrocyte precursor cells (OPCs). Failure of this remyelination program often occurs due to the improper recruitment of OPCs into injury sites, contributing significantly to ongoing neurological dysfunction and disease progression. Understanding the mechanisms controlling OPC biology during remyelination will provide insights as to why myelin repair fails in human cases. Importantly, OPCs dynamically produce primary cilia, microtubule- based organelles that transduce intercellular cues in a specialized signaling compartment. The role of primary cilia in regulating developmental pathways in OPCs remains poorly understood. Here, we show that OPCs require primary cilia to respond properly to WMI. First, this grant will demonstrate that genetically removing primary cilia from OPCs results in inadequate WMI repair, identifying the primary cilium as a critical effector of biological change in OPCs necessary for the WMI response. Furthermore, as there remains little mechanistic understanding of ciliary signaling pathways in OPCs, we will use a combination of approaches that ultimately define a GPCR/cAMP/CREB signaling axis beginning at the primary cilium as a crucial regulator of OPC biology. Finally, with recent advances in proximity-labeling, we can now catalogue the proteins that survey OPC primary cilia using a technique termed cilia-APEX. This grant will utilize cilia-APEX to identify signaling molecules that localize to OPC primary cilia in vitro and during remyelination in vivo. This will demonstrate dynamic changes in the protein content of OPC primary cilia during different stages of remyelination, while also adding significant insight into the extent of ciliary functions in OPCs. Together, these studies will show that primary cilia are a critical signaling module in OPCs for the regulation of remyelination, and will reveal potential therapeutic target for conditions such as MS and CP, where the OPC response to injury can be dysfunctional. 1

NIH Research Projects · FY 2026 · 2023-04

ABSTRACT In humans, an age-related decline in neuromuscular function is associated with loss of muscle mass (sarcopenia), increased frailty and a degradation of both health and quality of life. The only known treatments are life-style based, including exercise and diet. Rodent models have been used to demonstrate age-related changes at the neuromuscular junction (NMJ) including the fragmentation, remodeling and eventual denervation of muscle. As eloquently stated by Joshua Sanes and Jeff Lichtman, “A key feature of the adult NMJ is that it is remarkably stable under ordinary circumstances yet capable of remodeling” when perturbed by injury or age. “This combination of stability and malleability implies that synaptic maintenance is controlled actively, yet little is known about the underlying molecular mechanisms”. This is where we have recently advanced the field. In a recently published paper we demonstrate the power of homeostatic plasticity to preserve neuromuscular anatomy and function with dramatic effects on organismal health, behavior and lifespan. We refer to this as “Homeostatic Neuroprotection”. We propose to determine whether homeostatic neuroprotection counteracts the insidious effects of age-related neuromuscular decline over the normal lifespan of mice. This will be achieved by characterizing three independent mouse strains across the lifespan. We provide strong preliminary data supporting an evolutionarily conserved role for all three genes in the mechanisms of presynaptic homeostatic plasticity and accelerated aging in the mouse neuromuscular system. Age is one of the most important risk factors for nearly all neurodegenerative disorders. This line of research may underscore the general relevance of homeostatic neuroprotection as a future therapeutic avenue to ameliorate the adverse effects of age and neurological disease.

NIH Research Projects · FY 2025 · 2023-04

ABSTRACT Vitamin D deficiency is a widespread problem among the elderly and can precipitate sequelae that are particularly harmful to this vulnerable population. Hyperparathyroidism (HPT) is commonly associated with low vitamin D status, and the disruption in calcium homeostasis that is the cardinal feature of this disease can lead to a range of deleterious outcomes among the elderly, including loss of bone density, muscle weakness, and increased risk of falls and pathological fracture. As the primary endocrine organs responsible for hormonal control of serum calcium levels, the parathyroid glands are a known target for the actions of vitamin D, yet there are major gaps in our understanding of how vitamin D deficiency and attenuated vitamin D receptor (VDR) signaling contribute to the etiology and clinical presentation of HPT. The mechanistic intermediates linking the VDR to parathyroid hormone (PTH) hypersecretion in HPT remain unknown, and the current model of VDR- dependent expression of the calcium sensing receptor (CaSR) in the parathyroid does not account for the significant proportion of HPT cases where downregulation of CaSR abundance in parathyroid tissue is not observed. Recently, several key findings from our group suggest a testable new model for vitamin D-mediated actions in the parathyroid gland. First, we showed that the GABA B1 receptor (GABAB1R) can form heterodimeric complexes with CaSR, promoting tonic hypersecretion of PTH by opposing the coupling of CaSR with its obligate downstream G-protein effectors Gq/11 and Gi. Second, building upon a recent report demonstrating that soluble peptide derivatives of the amyloid precursor protein (APP) can bind and activate GABAB1R in neurons, we found that the APP-derived peptide Aβ1-42 can increase maximal PTH secretion by parathyroid tissue in a CASR- and GABAB1R-dependent manner. VDR expression and serum vitamin D levels are inversely correlated with the relative abundance of APP, Aβ1-42, the γ- and β-secretases required for Aβ1-42 production, and the phosphorylated form of the microtubule associated protein Tau (pTau). Functionally, ablation of APP in the parathyroid abrogates the development of HPT in VDR KO mice, and inhibitors of Tau phosphorylation can block the ability of Aβ1-42 to promote PTH hypersecretion. These data suggest that HPT driven by loss of VDR activity could arise at least in part through unregulated expression of Aβ1-42 and pTau. Based on these findings, we hypothesize that aging-induced increases in Aβ1-42-mediated signaling drive tonic PTH hypersecretion and that vitamin D deficiency exacerbates HPT disease severity by relieving suppression of Aβ1-42 production and Tau/pTau expression. To test this model, we propose three complementary, mechanistic specific aims: (1) to delineate the molecular actions of Aβ1-42 on CaSR, GABAB1R, and downstream signaling events that promote PTH hypersecretion; (2) to determine whether blocking the production or activity of Aβ1-42 and Tau delays HPT development in a murine model of CaSR insufficiency; and (3) to delineate the causal relationship between VDR and Aβ1-42/pTau signaling in the parathyroid. By defining the contributions of the CaSR/GABAB1R/Aβ1-42 signaling axis to PTH hypersecretion, this work will provide a clearer mechanistic understanding of the unexpected connection between β-amyloid peptides, vitamin D status, and parathyroid gland function.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY / ABSTRACT Bilingualism may protect against cognitive and brain changes in both healthy aging and Alzheimer’s disease and related dementias (ADRD) through enhanced executive functioning, arising from the need to constantly manage two or more languages. However, studies have yielded mixed results, and there remains an unknown role of bilingualism in 1) resistance to the development of neuropathology and in 2) resilience, the maintenance of cognition and/or brain structure in the presence of neuropathology burden. Further research to clarify the protective role of bilingualism in aging and ADRD is crucial and highly relevant, considering that over half of the world’s population and more than 20% of the US population speak two or more languages. This proposal will help clarify the role of bilingualism in healthy aging and in Alzheimer’s dementia (AD) by addressing the nuances underlying bilingualism, sociocultural factors, executive functioning, and dementia diagnoses. We hypothesize that bilingualism contributes to resistance and resilience through both network-specific (language and executive functioning) and domain-general effects, and its impact is moderated by bilingualism and sociocultural factors, type of executive functioning task, and AD variant. Our long-term goal is to develop a framework for bilingualism’s influence on resistance and resilience. We will leverage a well-characterized, prospectively studied cohort of monolingual and bilingual speakers at the UCSF Memory and Aging Center who span the range of healthy aging (N = 200) and AD variants (N = 400). This cohort is unique in its scope of longitudinal (2+ timepoints) neuropsychological, bilingualism/sociocultural, neuroimaging, and AD biomarker data. In Aim 1, we will determine which aspects of bilingualism contribute to a protective effect on executive functioning. We will group healthy aging and AD variant bilingual speakers based on bilingualism and sociocultural factors and assess for differences on cognitive measures and neuroimaging findings. In Aims 2 and 3, we will determine if bilingualism is associated with resistance and resilience by comparing monolingual and bilingual speakers within healthy aging and AD variant groups. We will test for differences between groups in grey matter (GM) volume and white matter (WM) integrity (Aim 2) and AD proteinopathy (Aim 3) and correlate the findings with performance on cognitive measures. We are well-suited to complete these aims given our exceptional multidisciplinary team with expertise in the neural underpinnings of language, clinical phenotyping of aging and AD, cognitive resilience, and multimodal imaging. Accomplishing these aims will significantly impact the field of ADRD research by give important insight into the neural basis of bilingualism and its influence on the trajectory of healthy aging and ADRD, thereby improving care for individuals with diverse language backgrounds.

NIH Research Projects · FY 2026 · 2023-04

PROJECT ABSTRACT. Lung cancer is the leading cause of cancer mortality worldwide, with non-small cell lung cancer (NSCLC) the predominant histologic subtype of lung cancer and lung adenocarcinoma the major subset of NSCLC. ALK gene rearrangements (e.g., EML4-ALK fusions) are validated targets in NSCLC and current ALK kinase inhibitors yield impressive responses. Despite this clinical progress drug resistance remains a problem that limits patient survival. Improved therapeutic strategies are critical to identify to improve clinical outcomes. We propose an innovative, multidisciplinary, and collaborative project to hopefully improve the survival of NSCLC patients by defining a new mechanism of oncogenic signaling that we uncovered by studying ALK fusion oncoproteins. We aim to capitalize on our discovery of membraneless cytoplasmic protein granules (condensates) as a distinct mechanism of oncogenic kinase signaling in cancer. Our data suggest an emerging paradigm in which certain ALK fusion oncoproteins, as well as other clinically-relevant oncoprotein kinase fusions such as RET fusions, form de novo their own phase separated protein-based subcellular compartment devoid of lipid membranes and utilize higher-order protein assembly as distinguishing principles underlying oncogenic output. These membraneless cytoplasmic protein granules comprise a mode of oncogenic signaling that is different from that of native receptor tyrosine kinase (RTK) signaling and oncogenic, mutant forms of other RTKs such as EGFR, which use classical lipid membrane-based signaling. The pathogenic biomolecular condensates formed by ALK (and other RTK) fusion oncoproteins locally concentrate the RAS activating complex GRB2/SOS1 and activate RAS in a lipid membrane-independent manner. RTK protein granule formation is critical for oncogenic RAS/MAPK signaling output in cells. We identified a set of protein granule signaling components and established structural rules that define ALK protein granule formation. For instance, protein granule formation requires the adaptor proteins GRB2 and SHC, in addition to the ALK fusion oncoprotein. Our findings reveal membraneless, higher-order cytoplasmic protein assembly as a distinct subcellular platform for organizing oncogenic RTK and RAS signaling in cancer. We propose 2 complementary Specific Aims using innovative methodologies to probe condensate biology to understand the role of phase separation in ALK fusion oncogenic signaling. We further define the protein architecture of ALK fusion protein granules and identify the key interacting proteins required for ALK fusion protein granule formation, oncogenic signaling and tumor growth. The proposed studies will establish a mechanistic understanding of RTK fusion condensate biology to lay a firm foundation for the future design of mechanism-based therapeutic strategies to interfere with ALK protein granule assembly per se and that complement conventional ALK-targeted clinical agents, which are ALK kinase inhibitors. This project will provide insight into this distinct form of oncogenic signaling with a focus on ALK, with broader implications for the understanding of condensate and RTK fusion biology and the design of differentiated treatment strategies.

NIH Research Projects · FY 2026 · 2023-04

Project Summary/Abstract Schistosomiasis-induced pulmonary hypertension (Sch-PH) is globally the most common cause of pulmonary arterial hypertension (PAH), and its classification as a “neglected tropical disease” underscores the large unmet need in understanding the disease pathobiology and developing effective treatments. Although accumulating data support the likely contribution of the transcription factor hypoxia inducible factor-1 (HIF-1) and HIF-1-regulated glycolysis to the pathogenesis of PH, their significance in Sch-PH is largely unknown. This proposal examines the potential role of HIF-1-dependent glycolysis in promoting Sch-PH, specifically in lung interstitial macrophages (IMs). Available data demonstrate increased pulmonary perivascular infiltration of IMs in both humans and experimental mice with Sch-PH; increased transcription of HIF-1-associated genes encoding glycolytic enzymes in murine lung IMs with Sch-PH; and a protective effect of HIF-1α deletion in LsyM+ myeloid cells against murine Sch-PH. Building upon these key insights, this proposal will test the hypothesis that HIF-1- dependent glycolysis in perivascular lung IMs critically contributes to the development of Sch-PH. Of the three potential growth-promoting mechanisms of glycolysis--lactate fermentation, the pentose phosphate pathway, and the mitochondrial Krebs cycle--the potential mechanistic link between lactate and pulmonary artery smooth muscle cell (PASMC) proliferation will be additionally examined. Two aims are proposed. Aim 1 will determine necessity and sufficiency of lung IM HIF-1α stabilization in Sch-PH by conditionally deleting and stabilizing HIF- 1α in IMs of transgenic mice. The effect of lactate dehydrogenase A (LDH-A; an enzyme responsible for lactate fermentation) deletion and oxamate (LDH-A inhibitor) treatment on Sch-PH severity will be examined. Aim 2 will spatiotemporally phenotype lung IMs, quantify their glucose metabolism, and test, using co-culture, if lung IM- derived lactate induces a pro-proliferative, pro-fibrogenic phenotype in PASMCs. Completion of the project will clarify how glucose metabolism in lung IMs contributes to Sch-PH pathobiology. The proposed career development plan was designed with the ultimate goal of supporting the applicant’s successful transition to independence as a clinician-scientist, in the field of pulmonary vascular metabolism. The plan leverages the combined expertise of his mentors and advisors in basic-translational research, their commitment to mentorship, and the collective resources for research and professional development at UCSF. In the five years of training, the applicant will acquire proficiency in key experimental approaches (spatially resolved proteomics; flow cytometry and cell sorting; Seahorse metabolic assay; mass spectrometry; and confocal microscopy), develop data analytical skills, disseminate his findings, and refine grantsmanship, collectively positioning him as an expert investigator in the field of pulmonary vascular metabolism.

NIH Research Projects · FY 2025 · 2023-04

Summary/Abstract: 30 lines Drug-resistant tuberculosis (TB) threatens our goal of ending the global TB epidemic by 2035. Early diagnosis of drug-resistant TB for timely initiation of appropriate treatment remains a challenge. Phenotypic drug susceptibility testing (DST) requires Mycobacterium tuberculosis (MTB) culture and takes 8-12 weeks. Molecular-based methods are rapid but require previously identified and validated resistance-conferring mutations. Unfortunately, mutations are not completely known for key drugs included in newer all-oral regimens. Our long-term goal is to develop a non-culture-based method for phenotypic DST that provides results in 48-72 hours from sputum collection and that can be used directly on sputum. Our innovative approach is based on MPT64, a protein secreted only during active growth that is highly specific for MTB. We demonstrated that MPT64 secretion in MTB cultures decreases significantly within 48–72 hours of anti-TB drug exposure for drug-susceptible but not drug-resistant MTB isolates. We have previously measured MPT64 directly in sputum samples with a limit of detection comparable with Xpert MTB/RIF Ultra (Cepheid, Sunnyvale, CA, USA). In this study we will validate culture MPT64 DST chemiluminescence-enzyme immunoassay (CLEA) for isoniazid (INH), rifampin (RIF), fluoroquinolones (FQs), bedaquiline (BDQ), linezolid (LZD), and delamanid (DLM). Our proposed approach will provide results within 72 hours of the time the culture is identified as MTB, compared to the 2-3 weeks required to obtain results from conventional DST (Aim 1). We will use the principles of culture MPT64 DST to develop a sputum MPT64 DST CLEIA, which will provide results in 48-72 hours from the time of sputum collection. We will identify protocols for sputum liquefaction, antibiotic exposure times, and CLEIA that result in the lowest limit of detection (LOD) for resistant MTB cells (Aim 2). We will prospectively evaluate sputum MPT64 DST in 860 patients with TB in the Philippines and in the country of Georgia (Aim 3). We hypothesize that both culture MPT64 DST and sputum MPT64 DST will meet or exceed minimum sensitivity (>95%) and specificity (>98%), which are the WHO high priority target product profiles for rapid DST. The innovative MPT64 DST assays will transform TB management by enabling early initiation of effective treatment based on rapid drug susceptibility data, thus improving patient outcomes and curtailing the spread of drug resistant TB that is critical if TB is to be eliminated by 2035.

NIH Research Projects · FY 2026 · 2023-04

Abstract: The goal of this proposal is to develop an engineered T cell therapy with potential of translation into human testing. We will develop a clinically optimized combination antigen sensing prime-and-kill circuit T cell for precise recognition and enhanced elimination of mesothelioma, a rare disease with poor prognosis. The work is built on our recent progress in T cell engineering and novel tumor antigen discovery: (1) a prime-and-kill dual antigen AND-gated circuit with fully human components (dubbed as SNIPR for SyNthetic Intramembrane Proteolysis Receptors) that facilitate clinical translation. (2) A novel tumor specific cell surface antigen ALPPL2 (aka ALPG) that is expressed in mesothelioma but not any of the normal human tissues except for the placenta. Paired with the credentialed mesothelioma antigen mesothelin, the ALPPL2 SNIPR → CAR circuit T cell enables precise temporal and spatial control of T cell activation at the site of the tumor, minimizes on-target off-tumor toxicity, reduces tonic signaling and T cell exhaustion, and maintains multifunctional T cell states. The circuit design is modular and flexible and can be induced to locally deliver additional immune modulatory payloads such as cytokines to further improve efficacy. In addition, our research has shown that SNIPR → CAR circuit T cells are capable of effectively targeting antigens that are heterogeneously expressed in tumors, a common pitfall for therapeutic efficacy. We propose to perform translation-enabling studies: (Aim 1) Optimize antibodies for SNIPR T cell construction, and develop biomarker for patient stratification. (Aim 2) Engineer and evaluate humanized clinical grade SNIPR → AND logic T cells in vitro and in vivo. (Aim 3) Evaluate SNIPR-engineered prime-and- kill circuit T cells in killing tumor with heterogeneous target antigen expression. Successful completion of the project will enable us to move the SNIPR → CAR circuit T cell to translational development and identify via the biomarker appropriate mesothelioma patients for clinical testing.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY/ABSTRACT Preterm infants are at risk for central nervous system (CNS) hemorrhage which can disrupt cerebellar maturation and lead to permanent neurodevelopmental impairment. The molecular signals in the disrupted neurovascular niche that block cerebellar development are not known. Thus, no therapeutics are available to prevent the developmental disabilities associated with preterm brain hemorrhage. Fibrinogen, a blood coagulation protein, crosses a leaky blood-brain barrier (BBB) and is a key driver of neuroinflammation, oxidative stress, neurodegeneration, glial scar formation, and inhibition of repair. We hypothesize that fibrinogen is a critical component of the neurovascular niche after BBB disruption that blocks cerebellar development in preterm infants. Our preliminary studies show: 1) Lipopolysaccharide (LPS)-induced systemic inflammation in neonatal mice increases vascular activation, fibrinogen deposition, and neuroinflammation in the cerebellum; 2) Fibrinogen depletion rescues cerebellar growth in systemic neonatal inflammation and plasma injection models of BBB disruption; 3) Fibrinogen inhibits neurogenesis from cerebellar granule neuronal progenitors (CGNPs) and is sufficient to disrupt cerebellar growth in vivo; 4) Fibrinogen activates the bone morphogenetic protein (BMP) receptor activin A receptor type I (ACVR1) in CNS progenitor cells to inhibit remyelination and neurogenesis; 5) Fibrin binds the CD11b/CD18 integrin receptor on microglia/macrophages to induce pro- inflammatory and pro-oxidant pathways that are toxic to CNS progenitor cells and impair regeneration; 6) fggγ390-396A knock-in mice, in which the binding site of fibrin to the CD11b integrin is mutated, have improved cerebellar growth during systemic neonatal inflammation. Our specific aims will test our working model, whereby fibrinogen deposition after BBB disruption alters cerebellar development through: 1) direct inhibitory effects on CGNPs via ACVR1 signaling, and 2) activation of innate immune responses via CD11b. In Aim 1, we will define the contribution of aberrant ACVR1 signaling to fibrinogen-induced cerebellar injury using CGNP-specific ACVR1 mutant mice and clinically relevant ACVR1 small molecule inhibitors. In Aim 2, we will determine the role of fibrin- CD11b-induced innate immune activation to cerebellar injury using fibrinogen mutant mice and a novel monoclonal antibody that blocks the interaction of fibrin with CD11b. In Aim 3, we will define how fibrinogen- ACVR1 signaling alters human cerebellar progenitor cell fate in induced pluripotent stem cell-derived cerebellar organoids using single cell transcriptomics. These studies will reveal molecular mechanisms at the neurovascular interface that link BBB disruption to CNS progenitor cell dysfunction in preterm infant brain injury. Thus, results from this proposal may open new treatment strategies to overcome the inhibitory neurovascular niche in preterm infant brain injury as well as other neurologic diseases with BBB disruption and fibrinogen deposition, such as multiple sclerosis, Alzheimer disease, and traumatic injury.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY Glioblastoma (GBM) is the most common primary malignant brain tumor in adults that is universally fatal despite multimodal treatment. Novel therapies are therefore critically needed. Immunotherapy, such as checkpoint blockade, has thus far failed to demonstrate clinical efficacy. GBM has high intrinsic resistance to antitumor immunity due to, among other factors, low expression of MHC-I and lack of T-cell infiltration. Strategies to promote immunogenicity of GBM to sensitize tumor to checkpoint therapy is needed. Protein phosphatase 2A (PP2A) is a ubiquitous serine/threonine phosphatase comprised of a catalytic (C), regulatory (B) and scaffolding (A) subunit. We have previously reported that pharmacological inhibition of PP2Ac can enhance the efficacy of anti-PD1 blockade in multiple preclinical models, including GBM. However, the mechanism(s) or cell type(s) responsible for the enhanced antitumor immune response is not well understood. Recently, we found that deficiency of PP2Ac, by genetic modification, in glioma cells resulted in enhanced interferon signaling, which is essential to eliciting antitumor immune response. PP2A deficiency in glioma cells enhanced MHC-I expression, tumor T-cell infiltration and sensitivity to checkpoint blockade in vivo. We also demonstrated that PP2Ac deficiency led to enhanced production of cytoplasmic double-stranded DNA (dsDNA), which is known to activate cGAS-STING signaling, a potent simulator of interferon production. Moreover, from unbiased screening of all known regulatory B subunits, we identified PPP2R2C, a specific B subunit of PP2A, to have a similar role as PP2Ac in promoting MHC-I expression. In this project, we will first elucidate the effect of PP2Ac deficiency in glioma cells on the immunological landscape of the tumor microenvironment and will identify the immune cell types responsible for PP2Ac modulated antitumor immunity. We will then dissect the molecular mechanisms that link PP2A deficiency to cGAS-STING activation and promotion of interferon signaling in the glioma microenvironment. We will also identify the role of the specific regulatory B subunit, PPP2R2C, in modulating dsDNA production and cGAS-STING signaling. Finally, we will investigate the ability of PP2Ac deficiency to enhance the therapeutic efficacy of radiation therapy, a major component of current standard-of-care and a known stimulator of cGAS-STING signaling. The immediate goal of this project is to identify the mechanisms of PP2A modulated immunogenicity in GBM, with the long-term goal of developing precise PP2A targeting strategies to increase effectiveness of immunotherapies for GBM. We believe this study fits the mission of NINDS to seek fundamental knowledge of the nervous system and to use that knowledge to reduce the burden of neurological disease such as brain tumor.

NIH Research Projects · FY 2025 · 2023-04

ABSTRACT Sleep dysfunction is highly prevalent and disabling across a wide range of neurological and psychiatric conditions. In neurodegenerative diseases, including Alzheimer’s and Parkinson’s disease (PD), disruption of sleep architecture has been linked to worsening of daytime motor and neuropsychiatric symptoms, as well as accelerated disease progression. It therefore also marks a major, untapped therapeutic opportunity. However, it is currently not known which cortical and basal ganglia structures and signals are responsible for disrupting physiological sleep rhythms in PD. The rationale of this proposal is that identification of the cortical-basal signals which disrupt sleep architecture in PD is an essential next step for developing sleep-specific neuromodulation therapies. To date, a critical barrier to progress has been a lack of chronic intracranial neural recordings during sleep in PD. This urgent need can be addressed by leveraging recent developments in sensing-enabled Deep Brain Stimulation (DBS), supporting longitudinal, high-resolution, multi-site, intracranial recordings in patients’ own homes. The overall objective for this proposal is to establish the pathological network dynamics that disrupt healthy sleep in PD and how they are modulated by DBS. Our preliminary work demonstrates abnormal cortico- basal beta (13 - 30 Hz) and gamma (60 - 90 Hz) oscillations across different sleep phases in PD. Our central hypothesis is that these pathological overnight neural rhythms disrupt physiological sleep signals, including slow wave activity (<4 Hz), and induce maladaptive network changes during sleep to impact daytime cortico-basal neural activity and connectivity. We will use sensing-enabled, closed-loop, DBS devices that are chronically implanted in a cohort of 16 PD patients, combined with interpretable machine learning techniques, to identify cortico-basal signal and connectivity changes during sleep disruption in PD. We will then evaluate causal mechanisms of cortico-basal oscillations by measuring waking connectivity using cortical evoked responses and through applying sleep-stage dependent closed-loop DBS. Bridging this knowledge gap will characterize the pathological network dynamics of sleep in PD and uncover key mechanistic understandings linking sleep rhythms to waking network activity. This will provide a foundation for the development of closed-loop DBS approaches that can restore normal sleep in people with PD. Following successful completion of the proposed research, we expect our contribution to have determined the principal pathological oscillatory cortico-basal dynamics of sleep disruption in PD. The proposed research is innovative, using new sensing-enabled DBS for longitudinal sleep recordings plus closed-loop neuromodulation to evaluate cortico-basal network models of sleep dysfunction in PD. This contribution is expected to be significant because understanding the fundamental neurophysiology of sleep dysfunction in PD represents a critical knowledge gap towards developing precision neuromodulation therapies for sleep, daytime motor / non-motor symptoms and disease progression in PD, which will also inform on other neurological and psychiatric conditions.

NIH Research Projects · FY 2026 · 2023-04

Project Summary/Abstract Acute respiratory distress syndrome (ARDS), the most severe form of acute lung injury (ALI), is common in COVID-19 and was a major driver of the now >1 million deaths in the USA since March 2020. Rigorous work has emphasized the key role the immune system and the microbiome plays in lung diseases like ALI. This includes an important role for pro-inflammatory T helper 17 (Th17) cells, which expand and increase their activation in lung tissue during ALI and lead to pulmonary fluid accumulation. While select gut bacteria can induce Th17 cells, the causal role of the microbiome in ALI and ARDS remains a major knowledge gap. Bifidobacterium adolescentis is a gut isolate with the highest Th17 inducing capacity of any human gut bacterium. My own preliminary data show that colonization of germ-free (GF) mice with B. adolescentis is sufficient to markedly alter the expression of many genes in lung tissue, including hundreds of genes implicated in ARDS. My data also indicates that B. adolescentis and other bifidobacteria secrete small molecules that activate Th17 cells, providing a plausible mechanism by which bifidobacteria could contribute to lung injury. My K08 application will build on a set of tools based on our laboratory’s recent discovery that the host ketone body, β-hydroxybuytrate (βHB), uniquely suppresses the growth of Bifidobacterium species. I hypothesize that Bifidobacterium, which are suppressed by gut epithelial βHB, secrete small molecules that reach lung tissue and activate Th17 cells during ALI. The proposed research plan will provide multiple training opportunities leading to a unique and sustainable research program. As a clinical fellow in the Turnbaugh lab (UCSF), I have learned microbiome data analysis, anaerobic microbiology, and gnotobiotic mouse husbandry. This project, coupled to in-depth support by lung experts Drs. Sheppard and Matthay (UCSF), will help me develop an independent research area that builds upon my past training. I will develop skills quantifying ALI severity in two ALI models and pair these ALI models with state-of-the-art tools in microbiome research. This proposal is both technologically and conceptually innovative. We will leverage resources available in our laboratory to selectively manipulate Bifidobacterium colonization levels in conventionally raised mice using βHB. My proposed experiments will help elucidate the metabolites produced by gut Bifidobacterium that induce pulmonary Th17 phenotypes, emphasizing the importance of considering bacterial metabolism for immune function and lung physiologic impairment. Our proposed studies will shift the focus to remote communication between prevalent human gut bacteria and lung tissues, mediated by secreted bacterial metabolites. For patients with ALI, this work will define putative mechanisms by which gut microorganisms contribute to devastating physiologic impairment and provide a conceptual basis to understand the scope and molecular impact of the gut microbiome on lung function in both health and disease.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY The overall goal of the UCSF Musculoskeletal (MSK) Training Program is to prepare Ph.D. scientists and M.D. or M.D./Ph.D. residents and fellows for a lifetime of scholarly pursuits that lead to in-depth understanding and improved care for patients with MSK diseases. We are confident that our proposed training program will accomplish this goal. Drawing on the solid mentorship skills of 35 nationally renowned and highly collaborative faculty with primary appointments in 10 academic and clinical departments, the UCSF MSK Training Program provides rigorous research training in basic and clinical/translational sciences important to the study of MSK diseases. These 35 participating faculty members bring multi-disciplined expertise that reflects areas of rapidly expanding research development such as stem cells, neurobiology, tissue regeneration, molecular physiology, and artificial intelligence. A vital aspect of the program is to focus exclusively on preparing postdoctoral trainees for independent research careers. The program proposes three slots for postdoctoral trainees, including two slots for Ph.D. postdoctoral fellows (research training duration of two years) and one slot for physician trainees from our research-track residency program in Orthopaedic Surgery (research training duration of one year). The central component of the training experience is direct research in the basic laboratory or clinical/translational research group of a primary research mentor. The training experience fuses this scientific training with sustained, individualized, and goal-oriented mentorship that guides trainees along the pathway to gaining their competitive research funding. The program includes formal coursework in ethics, rigor and reproducibility, statistical analysis, and grant-writing. It will also emphasize basic and clinical/translational science education by drawing on significant curricular activities designed explicitly for MSK science; innovative research experiences with an emphasis on scientific rigor and translation; individualized development plans that utilize evidence-based approaches for career planning; numerous exposures to state-of-the-art methods and techniques through weekly grand rounds and invited lectures, monthly research-in-progress seminars, journal clubs, and an annual retreat; and scientific communication through formal expectations of publications and presentations at national meetings. These activities capitalize on the programmatic and scientific infrastructure created by several existing MSK- related centers and institutes with UCSF-wide participation and influence. Overall, the training enabled by this program will attract and prepare junior scientists with the goal of understanding the mechanistic basis of MSK diseases, finding new therapies for these diseases, and ensuring that scientific discoveries lead to advances in public health in an equitable and cost-effective manner. In doing so, the program will ensure that a cadre of young scientists will pursue academic careers in MSK research and advance the field in fundamental ways.

NIH Research Projects · FY 2026 · 2023-04

PROJECT ABSTRACT Retinal vascular diseases are major causes of vision loss in the United States and around the world. To better treat these disorders, we need to understand the signaling pathways that control the growth and integrity of retinal blood vessels. Our recent publications and preliminary data detail a novel angiogenic signaling system centered around heme, a co-factor critical for oxygen transport, metabolism, and gene transcription. We found that heme promotes angiogenic growth in the retina by regulating tip/stalk selection, and that reduced heme production or import leads to reduced retinal vascularization and tissue hypoxia, similar to other retinal vasculopathies including retinopathy of prematurity, choroidal neovascularization, and the rare but important exudative vitreoretinopathies. Furthermore, we found that VEGF suppresses, while Norrin-bCatenin promotes, the expression of the obligate endothelial heme importer, Flvcr2. Based on these data, we hypothesize that heme, is involved in retinal angiogenesis and retinal vasculopathies. The Specific Aims of this proposal are to (1) determine how heme intersects with Notch signaling to control angiogenic tip/stalk selection, (2) determine whether induction of Flvcr2/heme signaling is sufficient and necessary to reverse the vascular defects and downstream vision changes observed in mouse models of exudative vitreoretinopathy, and (3) characterize the role for Flvcr2/heme in VEGF-induced angiogenic proliferation and neo-vascularization. To accomplish these aims, we developed new tools to directly manipulate heme in cultured retinal endothelial cells and assess heme transport and intracellular trafficking in vitro. We also generated new conditional knock-in and knock-out alleles to manipulate endothelial heme transport in vivo. Our studies will fundamentally impact our understanding of how endothelial heme levels are controlled, and the role of heme in retinal angiogenesis and vascular disease.

NIH Research Projects · FY 2026 · 2023-04

ABSTRACT Over the last decade, there has been a dramatic increase in deaths resulting from drug overdoses. Pharmacological interventions have proven to be transformative and life-saving in substance use disorders, particularly in opioid addictions and overdoses. Today, there is an increasing interest in having an armamentarium at clinician's disposal to address substance use disorders. There are three therapeutic classes of drugs that are highly sought: alternatives, attenuators, and antidotes. Having alternative therapeutic avenues for pain management other than opioids could help lower new addiction cases. For alternatives, we look to anesthetics that maintain analgesic effects at sub-anesthetic concentrations. For attenuators, we seek behavior-modifying drugs like ketamine and dizocilpine that have demonstrated reduced self-administration of drugs of abuse like cocaine, alcohol, methamphetamine, morphine, and nicotine in animal models. As a final line of defense, we endeavor to discover new drug overdose antidotes that reverse the toxicity of abused drugs. The discovery of novel chemical matter in these therapeutic classes could lead towards the development of new pharmacotherapies for treating addictions, but pharmacological modification of addictive behaviors in mammalian models is costly and challenging to evaluate. The objective of this proposal is to accelerate the discovery and characterization of novel small molecules affecting behavior using high-throughput screening of compounds in live animals guided by behavioral profiling as opposed to biochemical or cell-based assays. Our work exploits an automated technological platform in which the behaviors of hundreds of larval zebrafish under the influence of neuroactive compounds can be assessed and compared simultaneously. It enables the high-throughput screening of thousands of compounds for those that phenocopy neuroactive drugs of interest. The central premise of our approach is that pharmacological modulation by these therapeutic classes, their unique behavioral changes in larval zebrafish, and the identification of new related pharmacology are inter-connected. Accordingly, we predict that molecules that phenocopy anesthetics, ketamine, or the PQs ability to reverse benzodiazepine sedation will be new chemical matter for alternatives, attenuators, and antidotes of abused drugs. From this perspective, we can use the high-throughput behavioral assays in larval zebrafish as a primary screen of a structurally diverse set of >100,000 compounds. Importantly, from initial pilot screens we already have new chemical matter of all these classes in hand. We will perform mechanistic studies using state-of-the-art imaging of the zebrafish central nervous system to help further characterize new pharmacology. An essential part of the work is the translation of newly discovered neuroactives into rodent models of pain and substance use disorders.

NIH Research Projects · FY 2026 · 2023-04

Project Summary Neuronal-glial relationships play a critical role in the regulation of synapse formation and neuronal specification. The cellular and molecular mechanisms by which neurons and astrocytes communicate and coordinate are not well understood. The experiments in this proposal will uncover the molecular and cellular mechanisms by which neuron derived SHH protein can coordinate the cell behaviors of cortical astrocytes to facilitate the formation and function of neural circuits in the brain. In our studies we take advantage of the large complement of mouse genetic tools that allows us to specifically manipulate Shh signaling in astrocytes. Using cutting edge light and electron microscopy approaches we will define how specific interactions between astrocytes and neurons are dependent on Shh signaling and assess how astrocytes influence the development of functional circuits. Collectively these aims will provide a framework for understanding the delivery, reception and cellular response to SHH in the developing cortex.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY/ABSTRACT Background: The homeless population is aging, with an increasing proportion of individuals over age 50 who experience accelerated aging, high rates of mortality, and a high risk of not having their wishes honored at the end of life. Advance care planning (ACP) aims to elicit patients’ medical preferences; yet older homeless adults have low rates of ACP. Significant policy attention has focused on rehousing chronically homeless people into Permanent Supportive Housing (PSH), subsidized permanent housing with voluntary supportive services. Our previous research indicates that PSH may be the ideal setting to initiate ACP; however, no studies have engaged formerly homeless PSH residents in ACP. Our team has created PREPARE for Your Care (PREPARE) – an easy-to-use, evidence-based, online ACP program with video stories. This program includes easy-to-read advance directives, an ACP group visit guide, and an ACP one-on-one facilitation guide. Through a prior R34, we developed a Community Advisory Board (CAB) and together identified preliminary adaptations to PREPARE for the PSH setting. The project requires a final co-development process with formerly chronically homeless older PSH residents and staff and our CAB. The objective of this proposal is to co-develop PREPARE-PSH and compare the effectiveness of facilitated group versus one-on-one visits among formerly chronically homeless older adults in PSH. Aims: We will (1) co-develop PREPARE-PSH with PSH residents, staff, and our CAB; (2) conduct a Hybrid (NIH Stage III efficacy/effectiveness), Type 1, two-arm, cluster randomized trial comparing the effectiveness of two delivery strategies of the PREPARE-PSH program (i.e., ACP movies and easy-to-read ADs) – facilitated group vs. one-on-one visits; and (3) explore implementation- relevant factors associated with ACP engagement and sustainability of ACP in PSH. Methods: In Aim 1 we will engage in a rigorous co-development process through in-depth interviews and focus groups with 20 PSH residents and the CAB. We will use a theory-informed framework (i.e., Behavior Change Wheel (BCW)) and qualitative content analysis to co-create a PREPARE-PSH program. For Aim 2, the CAB noted that groups may be more feasible to scale vs. one-on-one visits and asked us to randomize by site so residents would not feel “left out” of the alternate intervention. Therefore, we will conduct a Hybrid (NIH Stage III efficacy/effectiveness), Type 1 cluster randomized trial comparing delivery of PREPARE-PSH through facilitated groups vs. one-on-one visits using mixed effects models. Randomization will be at the site level, balanced by site size. In Aim 3 we will purposively sample Aim 2 participants for in-depth interviews (n=40-50), conduct focus groups with PSH staff (n=40), and obtain input from CAB members (total n=15). We will explore implementation-relevant factors associated with: (a) high and low ACP engagement and sustainability of PREPARE-PSH using the BCW and the Consolidated Framework for Implementation Research (CFIR) model. Relevance to NIH and public health: PREPARE-PSH may reduce health disparities in ACP among formerly chronically homeless older adults.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY COL4A1 and COL4A2 mutations cause Gould syndrome (GS) – a multisystem disorder for which clinically heterogeneous cerebrovascular disease is the major consequence. Cerebrovascular disease in individuals with GS can range from porencephaly caused by germinal matrix hemorrhages in utero, to infantile seizures, to age- related cerebral small vessel disease (cSVD) and vascular cognitive impairment and dementia (VCID). Hallmarks of cSVD observed in individuals with GS include subcortical microbleeds, enlarged perivascular spaces, and lacunar infarcts. Importantly, Col4a1 mutant mice faithfully model patient phenotypes. Moreover, Col4a1 mutant mice have age-related cerebrovascular dysfunction including loss of myogenic tone and impaired hyperemic responses that are thought to be critical to VCID progression. The extracellular insults resulting from COL4A1 and COL4A2 are heterogeneous and complex, which represents a significant barrier to mechanism-based interventions. However, because GS is a devastating monogenic disease with a defined genetic cause, it is an ideal candidate for correction of the root cause of the disease via genome editing technologies. In this proposal, we will leverage vastly improved CRISPR nucleases, base editors, and prime editors along with novel viral vectors to test therapeutic approaches using primary GS patient cells and mouse models of GS. This project will provide important pre-clinical data to develop the first genome editing- based therapy for this severe monogenic disorder. The successful completion of this work could eventually provide a one-time, lifelong treatment that prevents both childhood stroke and age-related VCID for GS patients and create a roadmap for correction of similar diseases.

NIH Research Projects · FY 2025 · 2023-04

Abstract: This proposal, submitted in response to RFA-HS-22-001, in the priority area of “improving maternal health,” is prompted by long-standing racial and ethnic disparities in use of permanent contraception (historically called “surgical sterilization”). In the US, permanent contraceptive procedures are most commonly performed on women of color, women with chronic conditions such as diabetes, and those residing in rural communities. Although permanent contraception aligns well with some patients’ reproductive life plans, reports that 10% of women regret having undergone these procedures are troubling. These regrets and racial disparities are especially problematic given recent PCOR demonstrating that long-acting reversible contraceptives, such as hormonal IUDs are both more effective and less likely to cause pelvic pain than permanent contraceptive procedures. To disseminate PCOR about the comparative safety and real-world effectiveness of alternatives to permanent contraception, we will adapt the advance care planning (ACP) framework, which has been used to ensure that patients receive medical care when approaching their end of life that aligns with their personal treatment goals. This paradigm is a critical tool for supporting patients in making complex medical decisions, by emphasizing elicitation of values, communication skill building, and shared decision-making. By preparing patients approaching the end of their reproductive life to more effectively communicate to clinicians their personal values, priorities, and treatment goals, we will adapt the ACP model for reproductive life planning (RLP), with input from patients (Aim 1, focus groups) and clinicians (Aim 2, semi-structured interviews). We will then conduct an individual-level randomized clinical trial comparing this approach to disseminating PCOR relevant to discussions of permanent contraception with 300 women who identify as Black or have a chronic condition such as diabetes, and who wish to avoid future pregnancy (Aim 3). In evaluating this intervention, our experienced multi-disciplinary team will carefully examine patient-reported outcomes related to communication and satisfaction with care. We will also rigorously analyze heterogeneity of treatment effects by clinical and patient-level contextual factors to inform the potential need for future refinement of this ACP-RLP paradigm. By increasing equitable access to alternatives to permanent contraception, this project strives to increase the proportion of US women who are using a method of contraception that aligns with their personal preferences and reduce the proportion of individuals who regret having undergone a permanent contraceptive procedure.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY Significant disparities in maternal and infant mortality and morbidity exist in the U.S, and the MFMU Network aims to reduce the rates of preterm birth, fetal growth abnormalities, newborn morbidity, and maternal complications of pregnancy. Novel strategies, including new therapies and technologies, innovative study designs and interventions, attention to implementation science, inclusion of adequately diverse study participants into clinical trials, and input from the stakeholders impacted by health disparities are needed to improve outcomes for pregnant and lactating people. Although California accounts for nearly one eighth of annual U.S. births, the MFMU Network lacks a West Coast site. The University of California, San Francisco (UCSF) has a racially and ethnically diverse patient population, including many individuals with differing gender identities. The inclusion of historically marginalized communities, including segments of the population that are highly represented in California, will ensure the MFMU Network study results are generalizable, and will promote health equity for all persons. UCSF is a pioneer in innovative research techniques, such as genomics and metagenomics, as well as clinical research methods, including community partnership and implementation science. In addition, UCSF has a robust research infrastructure that has supported numerous translational investigations including epigenetics and infection studies, metagenomic sequencing in pregnant patients with obstetric complications, studies of environmental contaminants such as wildfire smoke on reproductive outcomes, and placental biology, including the impact of COVID-19 and COVID vaccines. UCSF has an extensive network of affiliated practices and referring hospitals that provide care for diverse patient populations with a myriad of pregnancy- related complications and together with its affiliate, Zuckerberg San Francisco General Hospital, performs nearly 4000 deliveries per year in mostly high-risk patients. Therefore, to support the mission of the MFMU Network, we will 1) Enroll a large and uniquely diverse, west coast participant population; 2) Contribute multidisciplinary expertise of UCSF investigators to high priority areas of public health that impact pregnant people and/or their infants, such as the COVID pandemic, the opioid crisis, and the impacts of climate change; and 3) Leverage our affiliation with the MFMU Network to develop a diverse group of young academic investigators. UCSF will bring a host of novel and innovative opportunities, including clinical populations and unique technological and research approaches that will strengthen the work of the MFMU Network.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY/ABSTRACT Glaucoma drainage device (GDD) surgery has gained popularity in managing patients with medically uncontrolled glaucoma. However, one of the major long-term complications of GDD is progressive corneal endothelial cell loss (ECL) leading to corneal decompensation, which requires complex care. Although surgical techniques have been modified to insert the tube into the ciliary sulcus to address this issue, anterior chamber (AC) tube placement remains the preferred location, as there is a lack of convincing data to validate the advantages of sulcus placement. For example, (1) Direct comparison of endothelial cell loss (ECL) after sulcus versus AC tube placement is limited to three retrospective studies; (2) Studies are inconsistent regarding whether intraocular pressure (IOP) and other surgical outcomes are similar after sulcus tube or AC tube placement; (3) Question remains whether a sulcus tube cause more AC microenvironment change due to higher chance of having chronic tube-iris touch than an AC tube, jeopardizing endothelial cell health, IOP control, or other surgical outcomes of the sulcus tube. To answer these questions, we propose a multi-center, outcome-masked clinical trial randomizing 240 subjects to sulcus tube versus AC tube placement. In this trial, we will compare ECL (specific aim 1), IOP control (specific aim 2) and AC microenvironment (specific aim 3) after GDD implantation with tube placement in the ciliary sulcus versus the anterior chamber. In the setting of increased use of GDDs, our proposed trial to identify better approaches to decrease its corneal complications is of substantial interest to both corneal and glaucoma specialists. Department of Ophthalmology at University of California San Francisco (UCSF), Francis I. Proctor Foundation at UCSF, University of Pennsylvania (UPenn), and 4 high-volume centers for GDD implantation will jointly execute this trial. The Department of Ophthalmology at UCSF will serve as Clinical Coordinating Center. UPenn will be the Data Coordinating Center. Francis I. Proctor Foundation will be the imaging reading center and metagenomic RNA deep sequencing analysis (MDS) center. This trial is innovative for a number of reasons including the randomization of surgery (sulcus tube versus AC tube) and application of novel MDS analysis (examining postoperative AC microenvironment), none of which has been prospectively studied before. It is aligned with the priorities of the NEI, studying high- resolution imaging techniques such as endothelial cell imaging and anterior-segment optical coherence to guide post-operative treatment and as potential surrogate trial endpoints in future trials. This world class team of collaborators have a proven track record for executing large NEI-funded trials in ophthalmology, and are well positioned to answer the important questions presented in this proposal.

NIH Research Projects · FY 2026 · 2023-04

ABSTRACT Passive transfer of immunoglobulins (IgG) against SARS-CoV-2 occurs from the mother to fetus by transplacental transfer and may protect the neonate against infection or cause disease. Recent studies have shown maternal SARS-CoV-2 infection leads to the dysregulation of infant immune responses in cord blood including altered T cell related cytokines and perturbations of immune cell subsets. It has not been yet shown if this immune priming is SARS-CoV-2 antigen-specific or if it is secondary to non-specific maternal or placental inflammation. Additionally, recent evidence has shown waning immunity after vaccination and further work is needed to understand protective immunogenic responses to SARS-CoV-2 epitopes to optimize future vaccination strategies in pregnancy, to benefit both mother and infant. We hypothesize that maternal SARS- CoV-2 infection actively primes fetal immune responses in utero through 1) the transfer of immune complexes with immunostimulatory activity, and 2) the passive transfer of both protective and autoreactive antibodies to the fetus. Further, we hypothesize that differential placental tissue responses correlate with differences in maternal- fetal crosstalk. In this application, we leverage valuable samples from two prospective cohorts of COVID-19 infection and vaccination in pregnancy to address the following specific aims: 1) Investigate the fetal T cell immunostimulatory potential of SARS-CoV-2 Ags transferred to the fetus in immune complexes through a novel mass spectrometry-based approach; 2) Map the antibody repertoire profiles of Abs generated after SARS-CoV- 2 infection vs vaccination to determine the breadth of maternal-fetal transfer of protective vs autoreactive immune responses using PhIP-Seq technology; and 3) Identify differential transcriptomic and proteomic responses to SARS-CoV-2 infection at the maternal-fetal interface that mediate differences in immune complex transfer through the application of laser microdissection on patient biopsies. These results have immediate relevance to understanding novel mechanisms of maternal-fetal immune crosstalk after viral infection. Our data will aid vaccination strategies to protect both the mother and baby against COVID-19 and may significantly advance in our understanding of maternal-fetal immune interplay in perinatal infections.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY This research plan is based on a strong scientific premise that hypoxic-ischemic (HI) brain injury induces brain arginase-1 (ARG-1) to exhibit selected neuroprotective functions, such as efferocytosis and regenerative scar formation. In our preliminary studies, we detected spatiotemporal changes in ARG1 expression and activity as a result of neonatal HI. We have shown ARG1 localized mostly in microglia at the injury site early after injury performing efferocytosis and persisted in the injury core at later timepoints in the area of the tissue scar. ARG1 inhibition decreased ARG1 efferocytic function and worsened histological outcomes. While ARG1 involvement in efferocytosis and scar formation in other organs is well documented, ARG1-dependent mechanisms of efferocytosis and scar formation in the neonatal brain after HI are unknown. I hypothesize that ARG-1 regulates efferocytosis by providing polyamines for cytoskeleton assembly, and efficient efferocytosis is a crucial process for regenerative scar formation where ARG1 provides proline for extracellular matrix formation. Using in vivo the Vannucci procedure (common carotid artery coagulation followed by exposure to hypoxia in P9 mice) to induce HI, I will test my hypothesis in the following Specific aims: I will define whether HI induces polyamine pathway in ARG1 microglia, and whether ARG1 inhibition translates to defects in cytoskeleton assembly and performance of efferocytosis (Aim 1). I will characterize how ARG1 signaling alters cell composition in the scar and production of the extracellular matrix (Aim 2). I will determine whether efferocytosis is necessary for scar formation, if changes in ARG1 signaling alter local immune response in the injury core and whether this impacts migration of progenitor cells to the scar and tissue remodeling (Aim 3). The aims will be conducted using novel techniques, such as TRAPseq and spatial seqFISH that will significantly improve our understanding of processes in individual cells and cellular transcriptome in 3D. The proposed project will significantly improve our understanding of arginase-1 pathway, efferocytosis and tissue regeneration as a modifier of brain hypoxic-ischemic injury. This project will also significantly advance my scientific growth through learning besides the basics of neuroimmunology, the multiomics approaches and advanced data analysis necessary for my independent research career. Dr. Ferriero and I selected outstanding mentors, that together with coursework and research plans are aligned to address my specific knowledge gaps to ensure my career development as an independently funded physician scientist and to apply for an R01 at the end of this proposal. The proposed experiments and timeline are within my capabilities and the capabilities of the laboratory, animal care, and UCSF facilities.