University Of California Los Angeles

NIH Research Projects · FY 2025 · 2025-01

ABSTRACT Transgender women (TW) in Vietnam face cultural and societal pressure, stigma, isolation, and violence, contributing to substance use, risky sexual behaviors, and suboptimal engagement in HIV prevention and care. Despite these health disparities and adversities experienced by TW in Vietnam, there are no TW-specific interventions designed to address their needs. TransAction is an evidence- and theory-based intervention, developed by Dr. Cathy Reback (Co-Investigator) and TW in Los Angeles County, comprised of four core elements: outreach, individual risk-reduction counseling sessions, skill-building and support groups, and social events. TransAction participation is associated with increased self-efficacy and social support as well as decreased HIV risks including engagement in exchange sex and substance use. TransAction is designed to be delivered in resource-limited settings, so it is well-positioned to serve as an evidence-based HIV health promotion intervention model for adaptation in Vietnam. Following the implementation science ADAPT-ITT framework, researchers from the University of California, Los Angeles, Friends Research Institute, and the University of Medicine and Pharmacy in Ho Chi Minh City (HCMC) have completed the first five steps (Assessment, Decision, Adaptation, Production, Topical expert) of adapting TransAction for TW in Vietnam. The four core elements of TransAction were deemed urgently needed while TW community members provided valuable feedback on the adaptation and implementation of TransAction for Vietnamese cultural competency. This R34 will build upon our strong existing U.S.-Vietnam and academic-community collaborative relationships to complete the last three steps (Integration, Training, Testing) of the ADAPT-ITT process. In Phase 1, the team will take a community participatory approach to work with TW community-based organizations (CBOs) to develop TransAction protocols, materials, and health/social service navigation plans in the context of Vietnam. TW peer facilitators will be identified and trained to deliver the intervention activities. In Phase 2, the adapted TransAction will be pilot-tested with 80 HIV status-neutral TW aged 16 and above from HCMC and adjacent provinces with a two-arm, randomized design. TW in the intervention condition will participate in individual risk- reduction counseling sessions and skill-building and support groups using an online/offline hybrid modality. Both intervention and control groups will be invited to social events. Intervention outcomes, including placement along the HIV prevention/care continua, multilevel/multifaceted stigma and coping, self-efficacy, service utilization, HIV risks, and general health will be assessed at baseline, 3-, and 6-month. Implementation outcomes, including feasibility, fidelity, and acceptability, will be evaluated based on data collected from multiple sources. This study will lay the foundation for a subsequent full-scale trial to evaluate the scalability, efficacy, and sustainability of the culturally responsive HIV prevention and care intervention for TW in Vietnam and other settings with limited trans-inclusive services.

NSF Awards · FY 2025 · 2025-01

Understanding ecological resilience (resistance to and recovery from disturbance) is critical as human impacts on marine communities accelerate. This is especially true on tropical reefs, where for half a century, communities have rapidly, and often unexpectedly, transitioned away from the desirable coral state. On healthy reefs, short turf algae replace corals after disturbance, but this is usually a temporary state before corals recover. Short turf is composed of fine hair-like filaments of highly nutritious algae that is targeted by many herbivorous fishes and allows for settlement of coral larvae during the recovery process. However, overfishing, nutrient pollution, and increased sedimentation may support the emergency of new seaweed communities that resist recovery, even if human impacts are reduced. This proposal addresses whether the order of human impacts that alter key ecological processes determines resilience of algal turf communities, a process that has never been evaluated. Field experiments are being performed in Moorea, French Polynesia, on a reef where novel seaweed communities have replaced corals over the last two decades. Both resistance and recovery of these emergent communities are being measured, as well as any processes or seaweed traits supporting these properties. The project provides research training for undergraduate students and supports a gap-year program for recent graduates from underrepresented groups. On healthy tropical reefs, loss of coral often results in dominance by closely-cropped algal turf that can ultimately recover to coral. However, fishing of herbivorous fishes and increased nutrient and sediment supplies from developing watersheds can produce ‘ecological surprises’—the emergence of novel algal states that resist a return to short turf, even after human impacts cease. Understanding resilience, defined as both resistance to and recovery from disturbance, of short turf communities is intrinsically linked to understanding the resilience of the novel emergent communities, which are often dominated by long turf or macroalgae. However, transitions among states, both forward and backward, are currently unpredictable, influenced not only by current environmental conditions, but also potentially by the order of human impacts that have altered key ecological processes, a theory that has never been evaluated. This project is interrogating the importance of the sequence of human impacts to community transitions and their resilience on tropical coral reefs. The team is 1) conducting a manipulative field experiment to examine the resistance of short turf communities by sequentially applying increased fishing pressure, enhanced nutrient supplies, and increased sedimentation rates in all possible combinations of order, with no-change and simultaneous change treatments as controls; 2) measuring recovery following termination of experimental manipulations to assess how the order of impacts affects resilience; 3) examining the mechanistic underpinnings of resilience of both the original short turf and the emergent macroalgal states by measuring stabilizing feedbacks in all experimental plots; and 4) for states that transition to macroalgae, relating measured algal traits to stabilizing feedbacks to generalize the results to other tropical reefs. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2024 · 2025-01

PROJECT SUMMARY / ABSTRACT Trichomonas vaginalis (Tv) is a protozoan parasite that causes the most common non-viral sexually transmitted infection (STI), trichomoniasis, world-wide. The rise in drug resistant strains of Tv, and an emerging appreciation of the link between asymptomatic Tv infections with inflammation-driven pathologies demands better understanding of this prevalent human infection. Notably, Tv infection has been linked to reproductive complications, increased susceptibility to HIV, increased incidence of cervical cancer, and increased aggressiveness of prostate cancer. Clinically relevant infection most commonly occurs in female reproductive tract (FRT) where it must contend with the unique mucosal immune environment of the FRT. Interestingly in contrast to other STIs, the detection rate of Tv is highest in peri- and post-menopausal women. Thus, understanding how Tv subverts the host cell immune response, with a particular focus on the unique immune environment of the FRT, will be key in attempts to prevent pathogenesis by the parasite. Over the past decade our lab has shown that extracellular vesicles secreted by Tv (TvEVs) are internalized by human cells where they can alter pathogenesis by altering cytokine secretion and increasing adherence of the parasite to its host cell. Our lab and others have also shown that neutrophils (polymorphonuclear cells [PMNs]), the major immune cell present at the Tv–host interface, kills Tv via a mechanism known as trogocytosis. Additionally, preliminary data in our lab has shown that TvEVs down-regulate expression of a non-canonical, type I interferon, IFNε and that pretreatment of prostate cells with IFNε is protective against Tv-mediated killing. Interestingly, IFNε, is known to play an important role in protection of the FRT from bacterial and viral STIs. To elucidate the role of IFNε and TvEVs we aim to: i) determine which step(s) in the IFNε-mediated type I IFN response is blocked by TvEVs and ii) determine the role of IFNε and TvEVs during Tv infection of vaginal epithelial cells and interaction with PMNs. The work proposed in this fellowship will increase our understanding of how TvEVs subvert an FRT specific immune response mediated by IFNε that would otherwise be protective against the parasite.

NSF Awards · FY 2025 · 2025-01

In everyday life, the properties of materials are often judged by how they react to simple actions like poking, prodding, stretching, and squeezing. Similarly, in scientific and industrial applications – from polymer recycling to 3D printing – measuring a material’s response to repeated pushing and pulling is a good way to predict its performance. Despite the usefulness of these tests, however, there is no universal way of to interpret their results. This project will show that systematically tweaking the deformation cycle and analyzing the resulting changes can lead to new insights into a material’s behavior in a broad range of processes. The scientific tools developed for this project will have implications in other engineering disciplines where analogous “push-and-pull” tests are applied, including signal processing, process controls, and electrical circuits. Finally, this award will support outreach efforts in STEM education for high school students. In partnership with programs at UCLA, a high school students from the Los Angeles and Inglewood Unified School districts will receive training in computer literacy and scientific computing, including applications related to the project's research. Large Amplitude Oscillatory Shear (LAOS) flows are widely used by industry for material characterization of complex fluids, from commodity polyolefins to next-generation 3D printing gels. In principle, LAOS data contains valuable information about the structure and composition of a fluid, but the quantitative interpretation of experimental LAOS measurements is still an active area of research. This project will generalize the concept of parallel superposition rheology to LAOS flows, allowing a complete characterization of the material’s linear response function for perturbations about a base LAOS deformation cycle. This linear response function can be formulated as a harmonic transfer matrix (HTM), and preliminary data suggests that the HTM can be useful for both fundamental material characterization and prediction of shear banding flow instabilities. Overall, our proposed research will deploy a combination of theoretical, computational, and experimental tools to formalize and validate this new approach to LAOS banding stability analysis, develop interpretive frameworks for quantitative material characterization, and explain distinctions in the underlying bifurcation structure of steady and oscillatory shear banding flow instabilities. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2026 · 2025-01

Macrophages play vital roles as sentinel cells of the immune system to coordinate immune responses in response to pathogen exposure. Macrophage responses are shaped by their exposure history. We and other have shown that when exposed to pathogens or cytokines, macrophages remodel the epigenome through formation of de novo enhancers. This epigenomic reprogramming produces innate immune “training” and alters responses to subsequent stimuli. “Trained” macrophages may have increased capacity to fight infection, but when dysregulated, may also produce unwanted inflammation. Thus, it is critical to develop a mechanistic understanding of how epigenomic remodeling and innate immune training occur. We have recently described how the transcription factor NF-κB produces de novo enhancers in a stimulus-specific manner (Science, 372, pp.1349-1353). Here, we extend those findings to study the roles of the interferon- regulatory factors (IRFs) and STATs, focusing on IRF1, IRF3, IRF7, ISGF3 and GAF. Their regulation is dynamic and interdependent and they all bind the same DNA sequence. Thus, how their functional specificity is regulated is not intuitive. Our studies focus on bone-marrow-derived and tissue-resident macrophages. HYPOTHESIS: IRF/STAT family members remodel the epigenome and chromosome organization of macrophages in a combinatorial manner; each factor plays a distinct role, working sequentially to reprogram gene expression responses to subsequent stimuli. In Aim 1 we will dissect the roles of innate immune IRFs in de novo enhancer formation, using H3K4me1 ChIP- seq and ATAC-seq after stimulation with a systematic panel of ligands and genetic knockouts. Based on our preliminary results we hypothesize that IRF1 is a critical pioneer factor. We will test this hypothesis in vitro and in vivo with peritoneal macs and Kupffer cells In Aim 2 we will dissect how IRF/STATE chromatin remodeling affects later gene expression in response to subsequent endotoxin challenge. We distinguish between persistently activated genes, potentiated gene responses, and tolerized genes, and identify distinct functions by IRFs in these regulatory strategies. In Aim 3 we will connect the de novo enhancers of Aim 1 and the gene expression training of Aim 2, by leveraging recent advances from the 4D Nucleome Consortium to predict how IRF-dependent de novo enhancers mediate large-scale chromosomal remodeling to move immune genes closer to or away from nuclear speckles, so-called transcription factories. We will test these predictions with CRISPR-edited IRF enhancers.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY/ABSTRACT This application details a five-year research and career development plan to facilitate the transition of Dr. Yajing Gao from a post-doctoral fellow to an independent faculty position. Dr. Gao received her PhD in Immunology from UT Southwestern Medical Center focusing on mucosal lymphocyte differentiation. She then completed 4 years of post-doctoral training in lipid metabolism and physiology at University of California, Los Angeles. The mentored phase (K99) of the award will be completed under the continued guidance of Dr. Peter Tontonoz, an internationally recognized leader in lipid metabolism with over 25 years of mentorship in post-doctoral career development. Dr. Gao’s transition to independence will be supported by Dr. Tontonoz, co-mentor Dr. Martín, and a diverse advisory team with scientific expertise related to the proposed research, as well as a track record of promoting trainees’ independence. The applicant has identified targeted training objectives, including technical and didactic coursework and conferences to overcome scientific deficiencies in gastroenterology and metabolic analysis, to improve professional skills in grant writing, project management, and mentoring, and to secure an independent faculty position. Under the exceptional institutional environment for metabolism research and career development at UCLA, Dr. Gao will have access to top-tier training resources, core facilities, seminars, and exceptional collaborators to achieve her career goals. Nutrient deficiencies often accompany chronic gut inflammation and immune dysfunction. Yet molecular insights pertaining to the immunological regulation of nutrient uptake are lacking. The applicant’s recent work identified a connection between hyperactive type-3 immune cells and intestinal metabolic imbalance. Preliminary studies demonstrate that the mucosal type-3 cytokine directly controls the nutrient-handling abilities of the enterocytes. Aim-1 will identify signaling components and the underlying mechanism by which type-3 cytokine modulates enterocyte functionality, and test a promising strategy to correct this metabolic deficiency. In Aim-2, the candidate will obtain training to establish a mass-spectrometry-based approach to measure longitudinal nutrient flux in the intestine accompanying dietary change and delineate the contributions of distinct type-3 lymphocyte populations. Aim-3 will investigate the functional significance of the cytokine receptor in enterocytes and test the impacts of its loss on systemic metabolism. These studies will uncover fundamental enterocytic pathways governing nutrient flux, which is sensitive to the immune microenvironment, and inform the therapeutic utility of these targets for various inflammation-related digestive disorders. Through this award, Dr. Gao will develop scientific and professional skills in order to become an independent investigator at the intersection of mucosal immunology and intestinal physiology.

NIH Research Projects · FY 2026 · 2025-01

Project Summary/Abstract Astrocytes are ubiquitous CNS cells that make extensive contacts with neurons. Astrocytes serve diverse roles, including ion homeostasis, neurotransmitter clearance, and contributions to neurovascular coupling, which position them as critical components of neural circuits that contribute to brain physiology and disease. Once thought of as homogeneous, recent studies have highlighted astrocyte heterogeneity across and within brain regions. As tools to study astrocytes have improved, one important goal is to determine if defined populations of astrocytes regulate behaviors associated with the brain region they are located in. An ideal population to explore this larger question is the newly discovered subpopulation of Crym+ astrocytes within the nucleus accumbens (NAc), a structure within the ventral striatum that is well characterized in the context of rewarding behaviors and addiction. The focus of this application is thus to determine the role(s) of Crym+ astrocytes in behaviors relevant to the NAc. Crym encodes the protein µ- crystallin that was recently identified in a specific population of central striatal astrocytes to regulate perseverative behaviors of mice. The current application seeks to elucidate the functions of Crym+ NAc astrocytes (~45%) using integrated molecular, cellular, and physiological assessments during natural behaviors and a model of fentanyl-evoked opioid use disorder (OUD). I will test the hypothesis that astrocytes in the NAc have separable properties relative to the dorsal striatum, contribute to goal- directed behaviors, and that they are altered in a clinically relevant model of opioid use disorder (OUD) where goal-directed behaviors are disrupted. This hypothesis is based on exciting preliminary data described in the application, including data showing that Crym expression is decreased in NAc astrocytes in the OUD model. I will test this hypothesis with three Specific Aims. Aim 1 will characterize basic biological properties of astrocytes in the NAc. Aim 2 will assess functions of Crym+ NAc astrocytes during goal-directed behaviors. Aim 3 will determine how NAc astrocytes are altered in a model of fentanyl- evoked OUD. The aims are logically related to address my hypothesis but are not interdependent so that success in any one is not contingent on the other. The completion of the proposed experiments will provide me with rigorous training in state-of-the-art methods and allow me to directly address the functions of a molecularly defined population of astrocytes in the NAc.

NIH Research Projects · FY 2026 · 2025-01

ABSTRACT Studying how cortical circuits mature is critical to understanding neurodevelopmental conditions as proper wiring of these circuits is important for perception and cognition. Initially, spontaneous neuronal activity in the developing neocortex is characterized by intermittent, brief bursts of synchronous network events that interrupt much longer periods of no activity. Network activity then undergoes a transition from network-wide synchronous events to the asynchronous, sparse neuronal firing seen in adults. This desynchronization of network activity is a robust phenomenon that is conserved across many species, including in humans; in mice, it occurs during the second postnatal week. Because sparse firing is conducive to greater computational coding power, desynchronization is a crucial step in neurodevelopment. However, the mechanisms driving cortical desynchronization are unknown. Understanding these mechanisms would provide insight into the maturation of neural circuits and, critically, into the origin of neurodevelopmental conditions (NDCs), such as autism, epilepsy, and intellectual disability. Indeed, cortical network synchrony during development is persistently elevated in some mouse models of NDCs, suggesting that the trajectory of desynchronization is different in those NDCs. There are two principal mechanisms that could underlie cortical desynchronization: 1) intracortical mechanisms, and 2) bottom-up mechanisms. Intracortical mechanisms, such as an increase in inhibition or a decrease in intrinsic neuronal excitability, could shift the network away from the runaway excitation needed to generate synchronous events. Alternatively, bottom-up mechanisms could directly drive cortical desynchronization via changes in the activity of upstream brain regions like the thalamus. This proposal will compare the primary somatosensory cortex (S1), which receives dense bottom-up input from the periphery via thalamic relay nuclei, to the secondary motor cortex (M2), a nearby cortical region that receives little direct input from the periphery, to elucidate the contributions of intracortical vs. bottom-up mechanisms. Specific aim 1 will test whether inhibition drives cortical desynchronization in S1 and in M2 by utilizing in vivo longitudinal 2-photon calcium imaging from postnatal days (P) 9 to 14 to assess how the trajectory of desynchronization changes when local inhibitory interneurons are chronically inhibited with chemogenetics during the desynchronization window. Specific aim 2 will determine the role of bottom-up mechanisms by testing whether disrupting synchronous thalamic activity can alter the trajectory of cortical desynchronization in S1 and in M2. Specific aim 3 will characterize the specific genetic changes occurring in S1 vs. M2 during desynchronization using single-nucleus RNA sequencing. Together, these aims will provide insight into how cortical circuits mature. Crucially, this proposal will investigate different regions of cortex—with vastly different inputs—to disambiguate the contribution of intracortical vs. bottom-up mechanisms in driving cortical desynchronization.

NSF Awards · FY 2025 · 2025-01

Tektites are a type of glass formed by meteorite impacts. This project will study how elements behave in tektites and volcanic glass from the Moon. Researchers will study how elements are lost or depleted in these natural glasses (quenched silicate melts). Using silicate melts makes this research more accurate than previous work that uses data from simple compounds. This research could improve methods for studying Earth’s resources, which supports the national interest in science and technology. Another major benefit of this project is its emphasis on education and inclusion. The project will help train students who are from underrepresented groups in science for future careers in the Geosciences. The research team will also create an educational module on how chemical elements behave in nature. This resource will become publicly available to students and teachers everywhere. Researchers will also update the tektite display in the UCLA Meteorite Museum, where every year tens of thousands of visitors will learn about the latest research findings. The goal of the project is to investigate how certain moderately volatile elements, such as alkali metals (e.g. potassium, rubidium), transition metals (e.g. copper, zinc), and sulfur-loving elements (e.g. lead, gallium, germanium), behave when silicate melts (melted rock) evaporate. We will use a special laser-heating technique with an aerodynamic levitation setup to conduct these evaporation experiments. The project has two main objectives: (1) Volatility Scale of Elements: To create a ranking of how easily different trace elements evaporate; and (2) Activity Coefficients in Silicate Melts: To determine how the activity (or reactivity) of these elements changes based on temperature, the type of melt, and gas composition around it. The project’s findings will help improve models that predict the volatility of these trace elements, which is key to understanding how volatile elements behave and change on Earth and other rocky planets. Earth, for instance, has lost a significant portion of these elements during its formation. Understanding when and how this happened will shed light on Earth’s water sources, the formation of its atmosphere and oceans, and possibly even the origins of life. The project will train students at UCLA, especially women and students from other underrepresented groups in science, preparing them for future careers in geochemistry and related fields. In addition, the team will create a new educational module on the history and current understanding of how chemical elements behave in nature, making this resource publicly available to students and teachers everywhere. The proposal also includes an update to the tektite display in the UCLA Meteorite Museum, where tens of thousands of visitors will learn about the latest research findings every year. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY This application for the Mentored Research Scientist Development Award will facilitate the principal investigator (PI)’s transition to independence dissecting host-microbe interactions in obesity-related metabolic diseases. Candidate: The PI is an experimental biologist with a strong background in gut mucosal immunology and infection biology. She did her postdoctoral training under the mentorship of Dr. Rodney Newberry, a mucosal immunology expert. Under Dr. Newberry’s guidance, the PI demonstrated the novel role of intestinal goblet cells in promoting immune tolerance to luminal antigens. Additionally, the PI has also demonstrated how enteric bacteria interact with intestinal goblet cells to modulate their function. She will leverage the skills gained during her training to characterize the dialogue between obesity-associated gut microbes and intestinal immune cells. Career Development Plan: The PI will execute this proposal under the co-mentorship of Dr. Newberry and Dr. Samuel Klein (a leader in the field of obesity research), advised by a team of scientific experts who also have experience in nurturing independent investigators. Washington University provides an outstanding environment, collaborators and cores that will foster the PI’s career development. This proposal builds on the PI’s prior experience and fills in the gaps in her training, providing her with the tools needed for independence. It includes the following objectives: (1) master techniques in macrophage biology (i.e., immune cell trafficking, phenotyping); (2) developing expertise in obesity and metabolic dysfunction; (3) training in microbial ecology; (4) immersion in bioinformatics; and (5) publishing manuscripts directly related to this proposal. Research Plan: The scientific premise of the proposal is that gut microbiota from individuals with metabolically unhealthy obesity (MUO) compared to metabolically healthy obesity (MHO) propel intestinal and adipose tissue inflammation. To investigate the role of microbial drivers on the onset of metabolic syndrome, the PI has established a model of colonizing genetically identical wildtype mice consuming a normal chow diet with stool specimens collected from well-characterized obese or metabolically-healthy lean (MHL) human subjects with known degrees of adipose tissue inflammation, glucose intolerance and whole-body insulin sensitivity. Preliminary studies demonstrate that mice colonized with MUO, but not MHO or MHL donor microbiota have glucose intolerance, higher serum insulin concentration and significant expansion of macrophages in the intestine and adipose tissue. By completing the proposed aims, the PI will address 1) whether gut microbes from people with MUO promote host inflammation and the onset of metabolic diseases, and 2) determine the contribution of monocuclear phagocytes in microbiota driven-metabolic dysfunction. In completing these aims, the PI will complement her expertise in mucosal immunology with rigorous training in macrophage biology, microbial ecology and metabolic dysfunction to becomes an independent investigator with the long-term goal of dissecting how microbes contribute to obesity-related metabolic disease, in alignment with the NIDDK mission.

NIH Research Projects · FY 2026 · 2025-01

ABSTRACT Opioid use disorder (OUD) and chronic pain (CP) are interrelated, both impacting the brain's reward, affect, and pain circuits. OUD is prevalent among CP patients, and the euphoric effects and negative reinforcement properties of opioids are significant factors contributing to an increased risk of abuse in this population. The neural circuitry of OUD has been extensively studied using animal models and human neuroimaging, and an overall reduced functional connectivity within the reward network has been found among CP patients and those with OUD. However, the dynamic changes in connectivity from acute opioid use, and how these differ for CP patients who are at risk of developing OUD remain largely unknown. To address this gap in knowledge, we aim to uncover how acute opioid use impacts neural connectivity by directly recording human neuronal signals from regions associated with OUD. These regions encompass interconnected and overlapping circuits involved in reward, affect, and pain processing, and include: amygdala, hippocampus, insula, orbitofrontal cortex (OFC), medial and ventrolateral and dorsolateral prefrontal cortex (mPFC, VLPFC, DLPFC), anterior cingulate cortex (ACC), sensory thalamus, and periaqueductal gray matter (PAG). To achieve this, we will leverage a unique ability to directly record local field potentials (LFP) from awake humans, who have electrodes implanted for clinical purposes in deep brain regions, that opportunistically overlap with areas implicated in OUD. These recordings will be obtained from two patient groups: 1) epilepsy patients implanted with temporary brain electrodes for seizure monitoring (for 1-2 weeks), covering reward and affect processing areas, and 2) deep brain stimulation (DBS) patients undergoing awake DBS placement surgery, covering pain processing areas. Both groups routinely receive opioids to manage surgical pain, and both includes a subset of patients who have chronic pain (CP). This enables us to compare neural activity during acute opioid administration across individuals with varying degrees risk for developing OUD. In this project, patients will rate their pain and mood levels on a scale (0-10) before and after receiving opioid pain medications, and the change in reward/affect (Aim1) and pain (Aim2) network connectivity will be assessed, and further explored across patients with varying degrees of risk in developing OUD (Aim3). Through this K01 award, Dr. Ryu will lead the project and receive valuable mentored training by an interdisciplinary team specializing in substance use disorder, neuromodulation, human brain signal processing, and computational modeling. By the conclusion of the K01 award period, this research will reveal the neurophysiological signatures within the addiction circuitry related to acute opioid use, offering valuable insights for the development of future neuromodulatory treatments for OUD, and will further guide Dr. Ryu toward achieving independence in this research field.

NSF Awards · FY 2025 · 2025-01

A prominent paradigm used in both mathematics and computer science is the “structure vs randomness'' paradigm. At one end, it aims to identify structure in mathematical objects that is useful for their mathematical analysis and for the design of efficient algorithms for their manipulation. At the other end, it aims to use randomness and random-like properties for the analysis of objects that lack structure. Randomness is a common mathematical tool used throughout mathematics and computer science. One can sometimes identify “random like'' properties of mathematical objects that are as useful as true randomness for their study and analysis. In the best-case scenario, randomness can be identified as the absence of structure, and the two notions of structure and randomness can be seen as complementary. The project will expand the bridge between mathematics and computer science and may have a practical impact on widely adopted algorithms. Integration of research and education is a key component of the project. This project focuses on a novel approach towards this “structure vs randomness'' paradigm, whose main goal is to obtain significantly improved quantitative bounds for many important problems in mathematics and computer science. At the core of this new paradigm are two new concepts: spreadness and mixing. Spreadness is a quantification that there are not too many elements in any structured subset of the ambient universe and mixing is a measure for how two objects behave jointly via some common operation. The project aims to develop both the theory and applications of this new approach. On the theory side, we aim to build a versatile framework that can both connect existing applications and extend them to new ones. On the applications side, the proposal highlights exciting potential applications across several fields - additive combinatorics, communication complexity, graph theory, and fast combinatorial algorithms. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2026 · 2025-01

Project Summary An important function of the retinal pigment epithelium (RPE) is to ingest and degrade outer segment (OS) disk membranes from the photoreceptor cells. This catabolic role in the turnover of the phototransductive disk membranes is essential for the viability of the photoreceptor cells, with defects in the underlying processes leading to retinal pathogenesis. The proposed research focuses on the first event: the ingestion of the OS tips. We have developed high-speed live-cell imaging of this dynamic process by primary RPE cells. 3D analyses show that following formation of an f-actin cup around the OS tip, transient foci of actin filaments form at the site of scission of the OS, indicating that OS tip ingestion occurs by trogocytosis, involving dynamic polymerization of actin. The first two specific aims will combine studies on primary RPE cells in culture and in vivo analyses to investigate molecular mechanisms associated with the actin polymerization in this ingestion. The third aim addresses the daily timing of OS tip ingestion by performing experiments in which this event can be quantified by isolating it from the effects of phagosome degradation on phagosome number. The proposed experiments challenge current assumptions about how the OS tips are ingested by the RPE, and the daily timing of this cellular process. They are expected to generate novel findings that will establish a more accurate fundamental understanding of mechanisms underlying OS tip ingestion.

NIH Research Projects · FY 2026 · 2025-01

Abstract Following myocardial infarction, the heart is infiltrated in a precise spatio-temporal manner first by neutrophils and then by cells of the monocyte-macrophage lineage followed by proliferation of resident fibroblasts and endothelial cells. Cell-cell cross talk has been recently considered to be an important reparative mechanism but little is known about how the myocytes themselves engage in such cross talk with non-myocyte cells that are recruited to the infarcted region. In this proposal, we highlight and investigate an unusual cross talk between macrophages and cardiac muscle cells in the infarcted heart. We show that cardiac macrophages that are recruited to the infarcted heart disrupt cardiac muscle metabolism and lead to decreased NAD pools that play a critical role in cardiac muscle energetics in the infarcted heart. Bone marrow transplantation experiments performed by us definitively demonstrate that deletion of a NADase on macrophages leads to superior post infarct heart function and superior cardiac cellular respiration. Finally using monoclonal antibodies targeting specific NADases on macrophages, we demonstrate that specific targeting of such proteins could serve as therapeutic strategies for augmenting cardiac muscle energetics and post infarct heart function. We have created a multi-disciplinary team with expertise in cardiac metabolism, in vivo isotope labeled metabolite tracing, nucleotide biochemistry and cardiac physiology and comprehensively investigate the cellular, molecular, and metabolic underpinnings of how bone marrow macrophages regulate cardiac muscle metabolism and energetics. Our proposal sheds novel insight into the hitherto unappreciated role of how macrophages regulate cardiac muscle metabolism after heart injury.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY Cellular senescence is a complex cellular state characterized by cell cycle arrest resulting from DNA damage or other cellular stressors. Senescent cells play key roles in multiple biological processes, including development, tissue homeostasis, and acting as an anti-cancer mechanism. Moreover, senescent cells have been causally linked to sterile inflammation and disease due to their ability to secrete inflammatory factors known as the Senescence Associated Secretory Phenotype (SASP). Interestingly, macrophages, key innate immune cells, are emerging as a key source of senescent cells in multiple settings including tissue regeneration, wound healing, cancer, atherosclerosis, Alzheimer’s, and in the aging process. However, much is not known about the basic biology of macrophage senescence, including what genes, and signaling pathways regulate the senescent state, what biomarkers accurately define them, and the underlying biology regulating their functions including the SASP. Therefore, this proposal outlines our research goals for the next five years, which revolve around gaining a deeper understanding of the cellular and molecular mechanisms underlying senescent macrophage biology. This understanding is not only crucial for unraveling the complexities of their functions but also for comprehending how they impact biological processes in normal tissue homeostasis, and disease states. Our research project intends to delve deeper into the molecular mechanisms that govern cell cycle arrest, focusing on the role of the tumor suppressors Cdkn1a (p21) and Cdkn2a (p16), as well as other cell cycle arrest genes. To achieve this, we will employ omics-based methodologies and utilize CRISPR-Cas9 technology to uncover the complex roles of these cell cycle arrest genes in regulating macrophage senescence, SASP expression, and metabolism. Additionally, we plan to investigate the non-canonical roles of the P53-p21-CCND2 pathway in metabolism, particularly its impact on mitochondrial functions and metabolic flux in senescent macrophages. An integral part of our research will involve exploring inflammatory and metabolic pathways, such as the cGAS- STING, mTOR, and NF-κB signaling pathways, that regulate macrophage senescence and SASP production. This will include an assessment of the role of the NADase CD38 in modulating NAD+ metabolism in senescent macrophages and their influence on other cell types. Finally, we will investigate the impact of genetic variation on senescent macrophage biology by utilizing The Hybrid Mouse Diversity Panel. This will enable us to assess senescent macrophage phenotypes derived from various strains of inbred mice and determine the genetic determinants that influence senescent macrophage biology. In summary, our comprehensive research strategy is designed to enhance our understanding of cellular senescence in macrophages and its broader impacts on multiple biological areas, including general homeostasis and disease. Additionally, this research aims to pave the way for developing targeted therapies for diseases impacted by cellular senescence.

NIH Research Projects · FY 2026 · 2024-12

SUMMARY Accurate pathologic diagnoses, such as a cancer diagnosis, are the cornerstone of quality patient care. Accurate diagnoses require pathologists to perform a complex series of perceptual and cognitive tasks including visual search, pattern recognition, and decision-making. New artificial intelligence (AI) and computer- aided diagnosis (CAD) technology shows promise for assisting pathologists and could improve diagnostic accuracy. Rigorously evaluating the ways CAD cues can influence clinicians’ behavior, positively or negatively, is necessary to ensure that CAD improves outcomes as intended without introducing unanticipated but dangerous downsides. Our prior work in radiology showed that CAD tools can capture attention, induce overreliance, and actually lead to worse accuracy when used in practice. We hypothesize CAD may exert similar effects in pathology if employed ineffectively. By studying the impact of CAD cues on a large sample of pathologists, the proposed research will inform best practices for deploying CAD tools into clinical practice and have a profound impact on cancer diagnoses and patient care. Our project will randomize 250 pathologists to the presentation of different types and timing of CAD cues in 3 phases of interpretation. We will examine the impact on pathologists’ interpretive behavior and accuracy of two different types of CAD cues (Feature cues vs. Feature + Diagnosis cues; Aim 1), and two different timings of CAD cue presentation (Immediately upon review of the case vs. Delayed until after completing initial review; Aim 2). Finally, we will leverage eye-tracking to obtain a more granular understanding of the perceptual and cognitive mechanisms underlying the effects of CAD on interpretive behavior and accuracy (Aim 3). Strengths of our application include: 1) our experienced, multidisciplinary team with a history of successful physician recruitment; 2) access to existing and uniquely well-characterized biopsy cases; 3) use of a machine learning algorithm that our team developed in prior NIH funded research to create different CAD cues; 4) established relationships with 12 clinical sites across the country for participant recruitment; 5) an innovative remote image viewing and tracking platform for data collection; and 6) a scientific advisory board comprised of industry partners and clinical experts who will provide their insight and expertise to ensure that our results are immediately actionable for clinical practice. The proposed work is innovative, pioneering, and timely, addressing fundamental questions that accompany the introduction of new AI/CAD tools into the clinic. Our findings will evaluate the impact of CAD and illuminate the cognitive mechanisms that underlie successes and errors that occur during the diagnostic process as physicians interact with CAD cues. The results will inform solutions to guide the successful implementation of CAD into clinical practice to enhance its benefits, mitigate harms, and optimize benefit for patients.

NIH Research Projects · FY 2026 · 2024-12

PROJECT SUMMARY This study will provide data and a well-manualized treatment that are critical for the planning and design of a subsequent clinical efficacy trial to elucidate the effects of combining psilocybin with an evidence-based psychotherapy for major depressive disorder. Background. Psilocybin has emerged as a therapeutic agent for a range of mental disorders, including major depressive disorder; however, more recent data suggest its antidepressant effect may not be as powerful as initially reported. While the drug has received the bulk of attention, its use within the scientific literature is always complemented by some amount of psychotherapy. In fact, some psychedelic experts hypothesize that it is the therapy enhanced by the drug (rather than the drug itself) that leads to therapeutic benefits. It is critical we identify the necessary psychotherapeutic components that lead to safe and effective psilocybin treatment. Specifically, there is a significant need for a protocolized psychosocial treatment that can be tested and optimized to adjoin psilocybin treatment for major depression. We have preliminarily protocolized and, for this application, collected initial supporting data for a psilocybin- assisted cognitive behavioral therapy (PA-CBT) for adults with major depressive disorder. PA-CBT includes the core safety elements of standard psilocybin treatment, two psilocybin sessions (10mg & 25mg), and 12 sessions of CBT. Specific Aims. We seek to conduct a feasibility clinical trial of PA-CBT in two phases. Phase I will involve refining and optimizing PA-CBT using participant and clinician feedback through an open trial of PA-CBT. Phase II will be a randomized, two-arm, fixed dose trial that will test the feasibility, acceptability, and adherence to PA-CBT. Both conditions will receive two doses of psilocybin (10mg then 25mg, separated by one month). Participants will be randomized (1:1) to either a 12-session PA-CBT or a 6-session standard psilocybin-assisted therapy (PAT) condition that involves supportive therapy and contains no elements of CBT. Primary outcomes include feasibility (recruitment and retention of participants), quantitative measures of patient and therapist acceptability and treatment adherence (i.e., therapist fidelity to the study’s treatment manuals and, in the PA-CBT condition, participant’s adherence to CBT skill practices). Feasibility. We have received the regulatory approvals for the Phase I portion of this award. Our team consists of experts in psychosocial clinical trials for depression and psychedelic medicine. We have trained our core study personnel and collected supporting data on five initial participants. Impact. In line with NCCIH funding priorities, the proposed research will answer critical questions about the acceptability and feasibility of combining standard psilocybin treatment with one of the gold-standard psychotherapies (CBT) for major depressive disorder and begin to uncover the initial effects by which CBT can influence participants’ depressive severity and psychosocial functioning. This work will lead to the development of a manualized, evidence-based psilocybin- assisted therapy that can be further studied, optimized, and broadly disseminated to clinicians.

NIH Research Projects · FY 2026 · 2024-12

Abstract Alzheimer’s disease (AD), characterized by amyloid beta (Aβ) plaque accumulation, remains a formidable challenge due to limited therapeutic interventions. Despite recent FDA approvals, Aβ targeting antibodies fail to significantly impact the clinical progression of cognitive decline and the treatment is associated with significant side effects. Recognizing these limitations, our project proposes a novel therapeutic approach leveraging the versatility and potency of Chimeric Antigen Receptor (CAR) technology, primarily known for its success in cancer treatment. We aim to develop a hematopoietic stem cell (HSC)- based CAR-Macrophage (CAR-M) therapy targeting Aβ aggregates while simultaneously reducing central nervous system (CNS) inflammation, a critical factor in AD pathology. Our preliminary work with CAR therapy against HIV demonstrates the potential for long- term efficacy, safety, and CNS trafficking of CAR-modified cells, laying the groundwork for this innovative AD treatment. The project is structured into two phases: the R61 phase focuses on developing and optimizing Aβ- targeting CAR-M for in vitro efficacy against Aβ aggregates and inflammation reduction, and investigate the safety and CNS trafficking of HSC-based CAR- M in vivo. The R33 phase aims to evaluate the short- and long- term efficacy of this therapy in mouse models of AD, assessing its impact on amyloid plaques, neuroinflammation, and behavioral outcomes. By combining the expertise of leaders in CAR therapy and AD research, this project represents a groundbreaking step towards a potentially transformative treatment for AD, addressing both the pathological hallmarks and the inflammatory environment contributing to disease progression.

NSF Awards · FY 2024 · 2024-12

Coming out of the most severe and destructive viral pandemic of the past 100 years, the importance of understanding how viruses “work” is clear. Most viruses – including polio, yellow fever, Dengue, and SARS, etc. – have RNA genomes that are quickly turned or “translated” into viral proteins in host cells that self-assembled into new virus particles called capsids. Elucidating how this process happens is a high priority for preventing and treating these infections. This project sets out to connect in vivo experiments carried out in live cells with in vitro experiments carried out in a test tube with purified viral capsid proteins and RNA genome. While test tube studies allow for full control of the types and numbers of components and solution conditions in which they are interacting, live cells studies, on the other hand, involve viral RNA and capsid proteins in the presence of many unknown components whose effects on RNA translation and self-assembly into capsids have not yet been determined. The fundamental understanding that results from this research will enhance the ability to develop anti-viral treatments. Graduate students will be trained in an inter-/cross-disciplinary range of physical, chemical, biological, and translational medicine concepts and methods. Active outreach efforts aim at enhancing interest and understanding of science amongst budding scientists and lay persons of all kinds will be conducted. This project will be performed by an international collaboration between five different research groups in the US and France, each specializing in different experimental and theoretical techniques and each having extensive experience with one or the other of the plant (cowpea chlorotic mottle virus [CCMV]) and mammalian (hepatitis B [HepB]) viruses under study. These viruses were chosen because how significantly they differ in their host cell and capsid structure, so that general principles of viral self-assembly can be established. It is the goal of this project to elucidate the differences between in vitro and in cellulo viral processes by progressively adding to RNA and capsid protein a series of molecules that play key roles in the viral “life” cycle, mimicking the crowded interior of the cell. Using cell-free cytoplasmic (ribosome-rich) extract, viral RNA will be translated into protein products and the time course of capsid assembly will be investigated by a combination of experimental techniques, including magnetic resonance, X-ray scattering, and fluorescence and electron microscopies. Coarse-grained molecular dynamics computations and phenomenological theory will be used to analyze these kinetic data and to compare with what is learned using the same experimental techniques applied to corresponding virus assembly in test tubes, where all concentrations and solution conditions are controlled. This collaborative US/France project is supported by the US National Science Foundation and the French Agence Nationale de la Recherche, where NSF funds the US investigator and ANR funds the partners in France. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2026 · 2024-12

ABSTRACT Our long-term objective is to define cellular pathways that regulate lipid flux and to elucidate their impact on cardiovascular disease. Although the connection between hypercholesterolemia and vascular inflammation has been appreciated for decades, the mechanistic underpinnings of this association remain incompletely understood. This application builds on recent work that defined mechanisms of nonvesicular lipid transport and identified pathways to regulate PM cholesterol abundance in experimental systems. We showed that the Aster transporter family facilitates the movement of cholesterol from PM to ER, and that loss of Aster function leads to PM cholesterol accumulation. But the role this transporter in vascular physiology is completely unknown. Preliminary observations have revealed that Aster-A is responsive to inflammatory activation of endothelial cells (ECs), and suggested that loss of Aster function modulates vascular inflammation through control of PM cholesterol availability. We have further discovered that cholesterol binds directly to endothelial PM proteins, including ICAM-1 and VCAM-1, pointing to a novel mechanism by which changes in PM lipid composition may determine inflammatory responses. Specific Aim 1 will identify novel cholesterol-responsive proteins in the vasculature using chemical biology. Provocative early results have revealed that multiple cell surface proteins involved in vascular inflammation bind cholesterol directly and are responsive to changes in PM cholesterol. We will define the spectrum of cholesterol-binding proteins in ECs in response to hypercholesterolemia and in settings of vascular inflammation. Specific Aim 2 will define molecular mechanisms whereby membrane cholesterol regulates inflammatory signaling in vascular cells. We will determine how changes in PM accessible cholesterol affect signaling by membrane inflammatory receptors and elucidate how the physical interaction of cholesterol with endothelial adhesion molecules regulates their abundance. Specific Aim 3 will elucidate the impact of accessible PM cholesterol on vascular inflammation and cardiovascular disease. We will use Aster-A loss-of-function models to manipulate accessible PM cholesterol in cultured ECs and in animals.

NIH Research Projects · FY 2025 · 2024-12

PROJECT SUMMARY Nearly every human malaise is enhanced by age. Given the rise of the aging population, there is the urgent need for treatments that improve organ function and block or reverse aging. Such rejuvenation of organisms, tissues, and cells has recently become possible through approaches such as parabiosis and in vivo cellular reprogramming. For instance, the transcription factors OCT4, SOX2, KLF4, and cMYC (OSKM) can reprogram somatic cells to induced pluripotent stem cells (iPSCs) and reset the epigenetic profile, one of the most dependable hallmarks of aging, of the starting somatic cell to a new embryonic state. In an exciting modification of the original iPSC reprogramming method, partial OSKM-mediated reprogramming approaches were recently developed that rejuvenate tissues in vivo. In the initial proof-of-concept investigation, the cyclic induction of OSKM in aged mice yielded notable outcomes including enhanced expansion of pancreatic cells, improved glucose tolerance, and accelerated muscle regeneration after injury. Subsequently, OSKM-induced partial reprogramming was successfully implemented in a cell type-specific manner in vivo. Furthermore, recent advancements in vitro have demonstrated that partial reprogramming of ‘old’ fibroblasts by OSKM induced a rejuvenated reprogramming intermediate, wherein somatic identity is preserved. This rejuvenation was quantified to be the equivalent of approximately 30 years, as determined by epigenetic clock measurements. Thus, faithfully reprogramming cells undergo progressive and continuous rejuvenation of the epigenome, long before the pluripotent state is achieved. Intriguingly, the rejuvenated reprogramming intermediates can revert to the fibroblast state upon withdrawal of reprogramming factor expression and maintain its youthful state. Additionally, crucial functional attributes such as collagen production and migration reverted to a more youthful state, indicating comprehensive cellular rejuvenation. Partial reprogramming by OSKM may therefore have broad implications in regenerative medicine. However, the feasibility of partial reprogramming by OSKM in clinical settings is limited due to concerns regarding its potential to induce cells with a tumorigenic potential. We hypothesize that the mechanistic understanding of how partial reprogramming by OSKM induces epigenetic rejuvenation will lead to the discovery of strategies that can be more safely applied in vivo to induce rejuvenation. To unravel the molecular mechanisms underlying rejuvenation by partial OSKM reprogramming, we will 1) map the changes in the transcriptome, chromatin landscape, transcription factor binding, and epigenetic age across the OSKM-induced partial reprogramming, and based on these data, 2) identify the gene-regulatory and chromatin networks of rejuvenation and regulators of age. We expect that these approaches will begin to define rejuvenation factors that can be applied to other models and provide critical insights into the determinants of epigenetic aging.

NSF Awards · FY 2024 · 2024-12

The ability of quantum hardware to outperform classical hardware for tasks like computation, communication, and sensing is known as quantum advantage. Quantum advantage promises society-changing benefits including fundamentally secure communication, improved biomedical sensing, and breakthroughs in material and drug design. To date, useful quantum advantage for computation has not been realized. Though there is reason to be optimistic for near term advances, it is widely accepted that useful quantum advantage in computing will require fault tolerance at scale -- an advance that is still beyond the reach of current devices. This project is aiming to accelerate the development of fault-tolerant quantum computation by co-designing error correcting codes and the hardware that will run them. Currently, many quantum error-correcting (QEC) codes have been discovered, and their mathematical structures are becoming better understood. Meanwhile, hardware with a few logical qubits has been demonstrated and, in some cases, a small gain in process fidelity realized. However, the demands on hardware for current QEC codes are severe -- either enormous numbers of qubits are required or native gate fidelities must be extremely high (often, both). By designing QEC codes to utilize the native gates and connectivity of the hardware, while simultaneously optimizing the hardware layout to support the QEC code, fault tolerance can be achieved with the minimum resource cost thereby delivering useful quantum advantage sooner. Of the existing hardware platforms, the trapped-ion QCCD architecture shows a special promise to achieve fault tolerance, especially in view of the recent progress where key problems such as qubit shuttling and wiring have been solved. Further, it currently provides the highest state preparation, measurement, and gate fidelities, and has led to commercial systems with the highest quantum volume. It also natively provides mid-circuit measurement with minimal cross-talk error and has a well understood error model. Finally, it provides all-to-all qubit connectivity. These features are likely to be necessary requirements for realizing fault tolerance in the near term. To accelerate progress in fault-tolerant quantum computing, an interdisciplinary team composed of computer scientists, engineers, physicists, and educators focuses on the identification of the most efficient QEC codes for the hardware and the optimization of their deployment on the QCCD system. Further, they research how to improve gate fidelities while working closely with commercial vendors to realize the assembly of such a fault-tolerant QCCD system. Finally, the team convenes a community of educators, industry and government leaders to chart the optimum route for meeting the workforce needs of the field. This project advances the objectives of Quantum Information Science and Engineering at NSF in response to the National Quantum Initiative Act for the continued leadership of the United States in QIS and its technology applications. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2026 · 2024-12

PROJECT SUMMARY/ABSTRACT This proposal is based on our successful SE-CRISPR editing of the APOE4 gene at the codon for amino acid 112 distinguishing it from APOE3 (1), in the brain E4-5XFAD AD model mice (2) by a single IV infusion of microfluidic synthetic exosome (SE)-encapsulated CRISPR sgRNA+CBE mRNA leading to increased E3 mRNA production in brain. The editing of neurotoxic major genetic risk factor for sporadic AD ApoE4 (E4) to benign E3 represents a powerful novel therapeutic approach for Alzheimer’s disease (AD) (3-6). Compared to E3, E4 exacerbates the two predominant hallmarks of AD brain, Aβ amyloid plaques and tau pathology (7-14). Utilization of SE-CRISPR base-editing technology that allows editing in both dividing and non-dividing cells to edit E4 to E3 even in a subset of cells may decrease the risk for AD conferred by E4. SE-CRISPR IV infusion therapy could compliment the currently approved antibodies for AD, aducanumab and lecanemab, which reduce Aβ pathology but only modestly reduce cognitive decline. For effective gene editing in AD, CRISPR components must be delivered across the blood-brain barrier (BBB). Our lab has developed a brain delivery platform comprising microfluidic reactor-synthesized deformable lipid nanovesicles the size of natural exosomes (<150 nm), called ‘synthetic exosomes’ (SEs), that can encapsulate therapeutic macromolecules. Our preliminary data demonstrate that SE-CRISPR crosses the BBB and edits E4 to E3 in the mouse brain. In this proposal, we plan to optimize E4 to E3 editing in brain and determine alterations in the E4 phenotype of AD models in which E4 has been edited. In Aim 1, the dosage/dose frequency of the current promising SE-CRISPR candidate for editing of E4 in E4-5XFAD mice would be optimized with a goal of >50% editing in brain. In Aim 2, synthesis of SE- CRISPR would be optimized by use of ionizable cationic lipids and surface modification with glycol(PEG)ylated lipids and transferrin (Tf)-peptides to enhance plasma half-life and BBB permeability, as assessed by determination of editing efficiency in E4-5XFAD mouse brain using NGS. In Aim 3, chronic efficacy and safety of prioritized SE-CRISPR candidate(s) will be assessed in SE-CRISPR vs. SE-empty IV-injected E4-5XFAD AD model mice (14-16 mice/cohort) 5 days and 3 and 6 months after dosing, including memory behavior at 3 and 6 months, using SE-empty injected mice as controls. Editing efficiency will determined by brain region (cortex, hippocampus, etc.) and cell type (neurons, astrocytes, etc.), and in liver and WBCs. Biochemical analyses will include brain/liver E3 and E4 mRNA and protein, Aβ40/42, phospho-tau, and inflammatory markers, and plasma neurodegeneration biomarker Nfl. In Aim 4, to show species translation, the effects of prioritized SE-CRISPR candidate(s) will be assessed in E4-knockin (KI) rats using the established dose/delivery schedule, compared to SE-empty controls (6-8 rats/group). Rats will be euthanized 5 days and 3 months post final IV tail vein injection, and E4 editing assessed by brain region/cell type, and biochemical analyses as described for E4-5XFAD mice to identify a SE-CRISPR candidate for further clinical development as IV infusion therapy for AD.

NIH Research Projects · FY 2025 · 2024-12

Project Summary More than 4 decades into the pandemic, HIV disease remains a considerable public health concern without a practicable cure. Drug-based therapy can control HIV but is costly, has severe side effects, and is not curative. Stem-cell based therapies have provided the only known cures for HIV infection, with three individuals functionally cured to date. However, replicating these successes has been challenging due to the high toxicities of treatment, need for transplant antigen matching, and requiring extensive myeloablation. However, these “cures” strongly suggest that immune system modification involving hematopoietic stem/progenitor cell (HSPC) transplantation can play a strong role allowing HIV clearance from the body. Gene therapeutic modification of autologous HSPCs to allow resistance of engrafted cells to infection along with the production of cells that are capable of targeting and eliminating HIV infected cells represents a more targeted approach to a HIV cure for all. Through systematic development, we have established an approach to direct cells to target HIV infection in vivo using a novel and optimized HIV-specific chimeric antigen receptor (CAR). This optimized HIV-specific CAR contains a truncated version of the HIV-binding region on the CD4 molecule (the D1D2 regions) with the CD3 zeta TCR signaling domain along with a 4-1BB costimulatory molecule signaling domain (termed D1D2CAR41BB). Extensive preclinical studies in humanized mice have determined that the D1D2CAR was superior in comparison to other CAR candidates in allowing the modification of HSPCs, producing lifelong safe engraftment, enabling development into mature CAR-expressing cells, and resulting in antiviral suppression of HIV. With this strong preclinical data suggesting the safety and feasibility of this approach, we seek to work towards investigational new drug (IND) development and clinical implementation of this approach. In this proposal, we plan to 1) Develop a comprehensive product development strategy for IND submission, 2) Formulate a strategic plan for clinical development, and 3) Establish a comprehensive plan for a regulatory strategy. By address these aims, we seek to develop a comprehensive IND-directed product development strategy to facilitate phase I clinical trials with this approach to attempt to functionally cure HIV infection.

NIH Research Projects · FY 2026 · 2024-12

PROJECT SUMMARY Realizing the potential of cell therapies for cancer requires advances in manufacturing, cell potency, and in vivo durability of anti-tumor responses. In vitro generation of T cells from induced pluripotent stem cells (iPSCs) can advance these goals by enabling complex gene editing in clonal, self-renewing, genomically validated “master” iPSC lines from which therapeutic T cells can be produced. This promise is tempered by the largely unknown objective function of these cells, particularly their in vivo persistence. Development of PSC-T cells as viable alternatives to autologous T cells therefore requires a mechanistic understanding of their responses to tumor encounter and, from this, clear strategies to achieve functional parity with PB T cells. In this proposal, we examine the hypothesis that autocrine/paracrine Fas/FasL signaling is a critical checkpoint limiting the persistence of antigen-specific PSC-T cells during serial antigen encounter. We base this hypothesis on our observations that: a) PSC-T cells exhibit a de novo effector/memory-like gene expression program, including expression of Fas and its ligand, FasL, despite expression of naïve surface markers; b) that Fas and FasL expression increases on PSC-T cells following serial antigen encounter; and c) that, even in the absence of tumor-expressed FasL, antibody blockade of FasL on PSC-T cells markedly enhanced serial tumor killing. To test this hypothesis, we will use the artificial thymic organoid (ATO) method, developed by our group, to generate mature, CD8+ TCR-T and CAR-T cells from human iPSCs. We will leverage the ability to generate clonal, isogenic iPSC lines with deletion of FAS, FASLG, or both, to interrogate the role each gene plays in PSC- T cell response to serial antigen exposure in a CD19 CAR-T model. This will include testing the effect of FAS/FASLG gene deletion on depth and duration of in vivo tumor responses and competitive fitness in a xenograft mouse model; and testing in vitro efficacy of FAS/FASL-deleted PSC-T cells against patient-derived B-ALL. As a potential strategy for therapeutic PSC-T cells development, we will use both immunocompetent syngeneic mouse models and long-term PSC-T cell engraftment in NSG mice to demonstrate the safety of Fas/FasL disruption with respect to autoimmune disease, uncontrolled proliferation, and transformation. Our goal in characterizing this regulatory pathway in PSC-T cells is to significantly advance our understanding of PSC-T cell anti-tumor function to further their potential as a promising T cell source for cancer therapy. Furthermore, mechanistic understanding of the role of autocrine/paracrine Fas/FasL in the antigen-specific responses of PSC-T cells may more broadly inform our understanding of this potentially important pathway in T cells from other sources.