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Fresno Regional Foundation

Fresno Regional Foundation ($150M in assets) supports nonprofits in CA through grants focused on education, health, arts, community development. Awards range from $2,000 to $75,000.

NIBIB - National Institute of Biomedical Imaging and Bioengineering

Applications of brain Positron Emission Tomography (PET) have been in place for over 40 years. The combination of quantitative PET systems with novel radiotracers has led to numerous imaging paradigms for understanding normal and pathological brain physiology and pharmacology. Brain-dedicated PET systems offer important advantages over currently available PET systems in terms of sensitivity and resolution. Funded by a BRAIN Initiative grant, we developed the NeuroEXPLORER (NX), an ultra-high performance brain PET system. To date, human results have produced exceptional image quality and the delineation of small brain nuclei. The system's performance derives from its long axial field of view, small crystal elements, depth of interaction measurement, and excellent time-of-flight resolution. This proposal takes the next steps to optimize system performance including accuracy and precision, expand on the capabilities for dynamic PET with tracer kinetic modeling, and demonstrate the real-world performance with a wide range of PET radiopharmaceuticals. The proposal includes the following Aims. Specific Aim 1: Optimize key aspects of the system performance and reconstruction methodology to maximize image quality and minimize noise. We will further optimize and improve rigid and non-rigid motion correction, investigate impact of detector crosstalk and corrections, and develop reconstruction methods with practical computation times. Specific Aim 2: Develop and extend the capabilities of the NX for dynamic analysis using tracer kinetic modeling. Our initial assessment of extracting image-derived input functions from the carotid arteries has been very successful, and we will further validate this approach against arterial blood samples with a large cohort study using multiple radiopharmaceuticals that are commonly used in research and have potential clinical impact. We will also develop and validate methods to estimate the radiopharmaceutical metabolite corrections using population modeling and kinetic analysis strategies. In addition, the ultra-high resolution and sensitivity of the NX allows the reassessment of optimal modeling methods, which will be explored for tracers with challenging spatiotemporal distributions. Specific Aim 3: Real-world performance of human brain scans. We will further develop our 3D printing methods to generate a full-brain phantom and use it to compare to other PET systems. We will facilitate access of this phantom to other sites with novel brain PET systems. To truly optimize NX human imaging, we will perform test/retest studies with multiple radiopharmaceuticals and compare NX images to those of a current state-of- the-art whole-body system (Siemens Vision). This work will demonstrate the significance of the NX's performance characteristics and enable harmonization with conventional PET/CT scanners. In addition, we will perform brain activation studies using functional PET in a paradigm targeting the olfactory system, which is highly relevant in the early manifestations of neurodegenerative disorders.

OD - NIH Office of the Director

The goal of this S10 Instrumentation Grant application is to acquire a state-of-the-art Cytek® Aurora Full Spectrum 5-laser with automated sample loader flow cytometer for the Flow Cytometry and Cell Sorting (FCCS) Core Facility at the Tulane University School of Medicine (SOM), housed and administered in the Department of Microbiology & Immunology. This instrument will support the research of a minimum of 12 NIH-funded investigators at Tulane SOM and serve as a research and educational resource for regional investigators. The Tulane SOM is an NIH-supported institution with a mission that integrates research, training and service in biomedical research to improve human health through basic and applied biomedical research. In this application we present research projects from 10 Major users and 3 Minor users whose work is performed at the Tulane SOM. These projects are focused on infectious diseases, immunology and inflammation, which are areas of expertise at Tulane SOM. This work attracts national collaboration. Each of the profiled users has a strong history of NIH funding and needs for advanced flow cytometry technology. The existing Cytek® Aurora (3 laser, 38 parameter) and BD LSRII (4 laser, 13 detectors) flow cytometer cell analyzers have been in continuous operation for a nearly decade. While these instruments were state-of-the-art when purchased, they are no longer sufficient to meet future needs with respect to high parameter capability, resolution and versatility. We propose purchasing a Cytek® Aurora equipped with an ultraviolet, violet, blue, yellow/green, and red laser permitting detection of 64 parameters and a high-throughput system. The Cytek® Aurora offers several advantages over our existing instruments to meet investigators’ future needs. The inclusion of multiple lasers ensures addition of at least 20 more parameters than are possible with our existing flow cytometry analyzers, which can detect up to 38 and 13 parameters respectively. Compared to other premiere systems on the market, the Cytek® Aurora offers the best overall system in terms of number of parameters, service and price. The availability of a high-parameter and versatile instrument such as the Cytek® Aurora is expected to attract additional investigators that require this technology, leading to new collaborations that will expand the scope of research at the Tulane SOM and the greater New Orleans area.

NIDCR - National Institute of Dental and Craniofacial Research

ABSTRACT Amyloid was identified in the context of pathology but does not always represent aberrant protein folding. Functional amyloid is now recognized in all kingdoms of life. Amyloid aggregates possess evolutionarily conserved cross -sheet quaternary structures with common biophysical properties enabling their detection and study. Microorganisms are now known to produce purposeful amyloid within biofilm environments, but, considering their prevalence, little is known regarding molecular events influencing amyloid formation. Dental caries is a biofilm-dependent disease caused by dysbiosis and overgrowth of acidogenic and aciduric bacteria, particularly Streptococcus mutans. Amyloid is observed in vivo within dental plaque. Our group was first to discover oral amyloids, and we have identified four amyloid-forming proteins in S. mutans. P1 (AgI/II), WapA, and Cnm are sortase-localized adhesins and virulence factors. The fourth protein, Smu_63, negatively regulates biofilm cell density and genetic competence. Extensive tertiary and quaternary structural characterization of P1 is in hand, with characterization of the other proteins underway. Our X-ray fiber diffraction evidence proved a classical stacked -sheet amyloid structure for S. mutans amyloids, and our work revealed a new paradigm for multiple streptococcal and staphylococcal amyloids in that naturally occurring adhesin truncation products play two key roles within the organisms' biofilm life cycles. First, in monomeric form by promoting adherence to cognate ligands via quaternary interactions involving the cell surface-linked parent proteins, and second in amyloid form by quenching adhesive function and apparently facilitating detachment of aging biofilm cultures. The left-handed Z-conformer of extracellular DNA was recently associated with bacterial biofilm stability whereas right-handed B-DNA is associated with detachment. Of interest, the amyloid but not the monomeric form of neuropathologic A drives conversion of Z- to B-DNA. Also, membrane lipids impact amyloidogenesis by an unknown mechanism. Cardiolipin-rich mitochondrial membranes modulate amyloidogeneis of Parkinson and Huntingtin Disease-associated -synuclein and Htt, respectively. Cardiolipin is a prevalent anionic lipid in S. mutans cytoplasmic membranes and extracellular membrane vesicles, particularly under stress conditions. In this application we will define reciprocal mechanistic influences of amyloid-forming proteins on B- and Z-forms of DNA in vitro and in vivo in mono- and multi-species biofilms (Aim 1), determine the impact of membrane lipid composition on amyloid levels during biofilm progression and assess mechanistic interactions of specific lipids of interest on amyloidogenesis of known virulence-associated proteins (Aim 2), and continue to use state-of-the-art methods including solution and solid-state NMR spectroscopy to identify and characterize structural transitions reflective of monomer to amyloid conversion and determine changes in amyloid signatures for each protein upon exposure to different DNA conformers, amyloid-modulatory lipids, and other amyloidogenic proteins (Aim 3).

NICHD - Eunice Kennedy Shriver National Institute of Child Health and Human Development

PROJECT SUMMARY/ABSTRACT Assisted reproductive technologies (ART) have revolutionized fertility treatments, yet implantation success rates remain suboptimal. Emerging evidence reveals that maternal tubal fluid, specifically extracellular vesicles (EVs), nanoscale membrane-bound carriers rich in bioactive molecules that play a pivotal role in embryo-maternal communication essential for early embryo development and implantation. EVs transport lipids, proteins, and regulatory RNAs, including miRNAs, which influence embryo physiology and developmental competence. Although in vitro studies demonstrate embryo uptake of EVs from reproductive tract cells across species, the physiological relevance of maternal EVs during natural embryo development in vivo remains largely unexplored. Critically, EV populations and cargo profiles differ markedly between in vivo and in vitro conditions, underscoring a pressing need to investigate EVs in their native environment. Our innovative study leverages oviductal epithelial cell-specific CD9-green fluorescent protein (GFP) reporter mice to directly visualize and track maternal EVs within preimplantation embryos in vivo. We have discovered CD9-GFP+ EVs localized in the perivitelline space of 4- to 8-cell stage embryos, providing the first direct in vivo evidence of maternal EV-embryo communication during early development. Building on this, our research pursues two complementary aims: (1) to comprehensively map EV distribution and profile miRNA cargo in oviductal and uterine luminal fluid throughout early pregnancy stages, illuminating dynamic changes in EV-mediated signaling; and (2) to elucidate the functional significance of epithelial cell- derived EVs by employing pharmacological inhibitors to disrupt EV biogenesis and release in vivo, assessing consequent effects on embryo development, implantation, and pregnancy outcomes. This study offers a significant advance in reproductive biology by uncovering the in vivo role of maternal EVs in supporting embryo development and implantation. Our innovative use of epithelial-specific CD9-GFP reporter mice to directly visualize EV transfer to embryos provides valuable new insight into natural embryo-maternal communication. By combining cutting-edge molecular profiling with functional inhibition of EV biogenesis in vivo, this research is uniquely positioned to identify key EV cargos that influence embryo viability and pregnancy outcomes. The results will deepen our fundamental understanding of early developmental processes and enable the development of novel, clinically relevant strategies to improve assisted reproductive technologies. Leveraging maternal EVs as biomarkers or therapeutic agents holds strong potential to enhance ART success rates and promote healthy pregnancies, addressing critical challenges in fertility treatment.

NIAID - National Institute of Allergy and Infectious Diseases

PROPOSAL SUMMARY Autoimmune diseases remain challenging to treat due to a lack of therapies that specifically target immune dysregulation without inducing generalized immunosuppression. Dysregulated immunoregulatory networks drive autoimmune disorders and are often associated with imbalances in regulatory T (Treg) and effector T (Teff) cells. Treg cell therapy or selective expansion of Treg cells in vivo can shift the Treg/Teff balance towards Treg cells to resolve ongoing inflammation and promote tissue repair to restore function and improve overall quality of life. However, selective and functional expansion of Treg cells during ongoing autoimmune conditions (i.e., therapeutic use) has been challenging. This is mainly because immune regulatory networks are complex, and our knowledge of the cellular and molecular choreography of Tregs in tissues is still evolving, particularly in the context of autoinflammatory disorders of the central nervous system (CNS). Treg cells hold tremendous potential to not only stop the course of the disease but also to restore neuronal function by inducing the repair of damaged axons. Therefore, our main objective is to investigate key principles of expansion of Treg cells to limit the pathophysiology of CNS autoimmunity and promote myelin repair. Capitalizing on the multiphoton imaging, state-of-the-art ratiometric calcium indicator, Salsa6f, label-free detection of myelin lesions, we aim to identify the role of combinatorial activation of T cell receptor (TCR), IL-2 signaling, and TGF-beta signaling for expansion of Treg cells in vivo. Using immunomodulatory Treg expanding biologics (TREBs) and experimental autoimmune encephalomyelitis (EAE), a model of MS-like disease, we will determine target engagement, biodistribution, and immunosafety of combinatorial Treg-expanding biologics (Aim 1); and define cellular and molecular determinants of Treg expansion and evaluate therapeutic efficacy in models of CNS autoimmunity (Aim 2). We postulate that selective expansion of Treg cells is an ideal strategy to curb ongoing autoinflammatory responses while preserving the immune system’s ability to fight new infections and promoting tissue repair for functional recovery. Although our exploratory/developmental project aims to establish a mechanistic link between Treg expansion and clinical improvement during CNS autoimmunity, in a broader context, our studies will guide the rational design of Treg-targeting immunotherapies, accelerating translation into effective and safe treatments for MS and several other autoimmune conditions.

NIMH - National Institute of Mental Health

PROJECT SUMMARY Somatic mosaicism, the genomic differences among the billions of cells in the human brain, may explain the incomplete penetrance and variable expressivity in highly heritable neurodevelopmental disorders. Thousands of clonal somatic mosaic variants (SMVs) in subpopulations of neurons have been discovered in brains of schizophrenia and autism patients, necessitating an urgent, unmet demand to determine if these diverse somatic mutations have a causal role in disease. Major challenges include (1) the inability of using conventional statistical methods for common variants to associate disease status with risk variant, (2) the vast space of non-coding candidates with unknown function, and (3) the unresolved relevant cell types and developmental stages linking mutations to phenotypes. Just as integrating high-throughput genomic-, CRISPR, and stem cell-based technologies resulted in significant progress in understanding germline risk variants, they represent a novel approach to uniquely address the major challenges in the field of somatic mosaicism. As a co-mentored computational and experimental biologist, I will leverage state-of-the-art functional genomic technologies and bioinformatic pipelines to systematically characterize all brain non-coding SMVs discovered to date, resolving their causal roles in neurodevelopmental disorders. From all SMVs identified in case and control brains, I will first create a functional catalog of expression-modulated SMVs in a developmental- and cell-type-specific manner by applying massively parallel reporter assays in human induced pluripotent stem cells (hiPSCs)-derived neural progenitor cells (NPCs) and post-mitotic neurons. By doing so, I will be able to interrogate whether differences in patterns of expression-modulated SMVs exist between cases and controls. Second, I will compare the somatic and germline genetic architectures across neurodevelopmental disorders, determining whether somatic mutations act via the same pathways as germline mutations, or affect genes relevant to diseases, indicating a causal role. By simultaneously uncovering the downstream transcriptomic profiles of hundreds of regulatory elements harboring SMVs with CRISPR screen, I will be able to pinpoint putative disease-causal SMVs. Finally, I will validate the phenotypic impact of putative causal SMVs in physiologically complex and relevant models including 3D brain organoids and “mosaicism-in-a-dish”, testing both cell-autonomous and non-autonomous mechanisms of SMVs. Overall, this work, representing a novel application of scalable functional genomic technologies to SMVs, provides a framework to identify SMVs with putative causal effects in neurodevelopmental diseases, advancing our understanding of a poorly understood disease mechanism. This fellowship will provide me with training encompassing computational genomics, stem cell models and broadly applicable phenotyping techniques, setting a foundation for me to launch an independent research program distinct from my mentors', querying somatic mosaicism's impact into novel cell types, contexts and diseases towards discovering novel therapeutic targets.

NINDS - National Institute of Neurological Disorders and Stroke

Project Summary Hypersensitivity to touch is common in neurodevelopmental disorders including autism spectrum disorder (ASD). Individuals with ASD can have aversive responses to certain textures or human touch, which can lead to emotional distress, social withdrawal, and difficulties accomplishing everyday tasks. In humans and rodents, aberrant touch sensitivity perinatally is predictive of ASD-linked traits later in life. The molecules and mechanisms underlying touch sensation are just being elucidated, and little is known about how they relate to ASD. Touch sensation begins with the activity of mechanically activated PIEZO2 channels that transduce forces into electrical signals. The overall objective of the proposed research is to investigate the role(s) of the Ca2+ sensing protein caldendrin in regulating PIEZO2, touch sensation, and neurodevelopmental processes controlling cognition/affective behaviors. Our project builds on our discovery that mice lacking caldendrin (Cabp1 KO) display tactile hypersensitivity and increased activity of PIEZO2 in dorsal root ganglion neurons (DRGNs). Neurite outgrowth, which depends on PIEZO2 as well as Cav1 L-type Ca2+ channels, is abnormal in Cabp1 KO DRGNs and differs in cultures from males and females. Moreover, Cabp1 KO mice exhibit ASD-like phenotypes, such as anxiety and anti-social behavior, which are more severe in males than females. Considering that genetic silencing of some ASD-related genes in DRGNs of neonatal mice causes tactile hypersensitivity and ASD-linked behaviors in adulthood, our central hypothesis is that caldendrin modulates PIEZO2, Cav1, and the structural maturation of DRGNs early in development, which has sex-specific effects in driving synaptic plasticity in the brain and cognitive, affective, and social behaviors in adulthood. We will use state of the art methods in biochemistry, electrophysiology, and optical imaging to test the following specific aims: (1) elucidate the mechanism whereby caldendrin modulates the activity of PIEZO2; (2) define the contributions of caldendrin to sex- specific patterns of neurite development; and (3) determine how loss of function of caldendrin leads to aberrant cognitive/affective behaviors.

Fundacao Calouste Gulbenkian

Portuguese foundation supporting art, charity, science, and education internationally.

NICHD - Eunice Kennedy Shriver National Institute of Child Health and Human Development

PROJECT SUMMARY / ABSTRACT This proposal is for the graduate training program at Vanderbilt University that is structured to support the early phases of neuroscience predoctoral education and training. In support of the overall NIH mission, the overarching objective of the program is to provide an exceptional training environment for the next generation of neuroscientists, and is built on the foundation of a strong training faculty with exceptional records of scholarship, research support and graduate mentoring. The heart of this mission is expressed in the academic and research goals of the program, which are to provide our students with a strong didactic foundation in the neurosciences through our core curriculum offerings, and to provide them with the opportunity to carry out state-of-the-art neuroscience research in the laboratories of a group of highly successful and committed mentors. In addition, the program has strong emphases on professional development, quantitative literacy and rigorous science, with the objective of building the requisite skills needed for success in graduate school and beyond, and of training an inclusive cadre of future independent investigators in neuroscience research. The Neuroscience Graduate Program at Vanderbilt is an interdisciplinary program that encompasses 5 different colleges and schools and 23 departments. Traditional and emerging areas of research strength in the program include: addiction, attention, brain evolution, cell signaling, cognitive neuroscience, circadian rhythms and sleep, CNS drug development, development, developmental disabilities, molecular genetics, neurodegeneration and neurotoxicity, neuroimaging, plasticity, psychiatric illness, sensory and multisensory systems, synaptic transmission, and vision. The program is currently home to 67 trainees and 73 training faculty. The proposal requests support 5 slots and provides the rationale and justification for this request.

National Park Service

1. Create a science and technology showcase for art conservators to expose more conservators to scientific methods. The showcase will lead to better conservation of cultural heritage. 2. Create a research opportunity for youth, students, and new professional conservators to be exposed to and undertake research in an art conservation science laboratory. Research opportunities will lead to a new generation of conservators with an understanding of research within the art conservation practice. 3. Disseminate preservation technology information.

National Park Service

The intern will be part of the Cultural Landscape Inventory team for the National Park Service and based at the Olmsted Center for Landscape Preservation in Boston, Massachusetts. Responsibilities and learning opportunities include: creating field maps to document cultural landscape characteristics and features, field measurements and photography, meetings and interviews with park staff, archival research for historic photographs and maps, writing landscape descriptions, preparing landscape management and maintenance recommendations, and formatting reports for distribution. These responsibilities will be carried out with the guidance of Olmsted Center staff, including landscape architects and horticulturists. The intern will also collaborate with other interns and the Olmsted Center's youth education program. The intern should have skills and enthusiasm for reading and writing; skills in communicating and working as a member of a team; an interest in landscape architecture, horticulture, history, and historic preservation; and an interest in learning graphics programs, including Adobe Illustrator and Photoshop. No previous experience working in digital graphics is necessary, but an interest in art and drawing is helpful. This position is NOT eligible for AmeriCorps.

Patrons of the Trumbull Nature and Arts

FY25-ARPA-Sub Recipient

Bridgeport Arts & Cultural Council

FY25-ARPA-Sub Recipient

NCI - National Cancer Institute

PROJECT ABSTRACT/SUMMARY Neoplasms of the central nervous system (CNS) are the most frequently encountered solid tumors of childhood and remain one of the top causes of death in children. Pediatric high-grade gliomas (pHGGs) and atypical teratoid rhabdoid tumor (ATRT) are particularly aggressive pediatric CNS (pedCNS) tumors associated with poor outcome. Therapy consists of extremely burdensome multi-modal treatment protocols with toxic profiles that cause patients to suffer from detrimental effects, which significantly impact the quality of life during their already limited lifespan. New therapeutic strategies are badly needed to increase the survival benefit and quality of life of patients with pHGG and ATRT. To address this need, we performed initial studies that demonstrated that primary CNS cancers display dysregulated iron homeostasis and that gallium maltolate (GaM), an iron mimetic metallocompound, inhibits the growth of pHGG and ATRT cells in vitro and in an orthotopic rat model, extending overall survival. Cancer iron metabolism is an attractive target for therapeutical intervention. Iron plays a vital role in the pathobiology of many cancers, including brain cancer. Gallium acts as an iron mimetic, enabling it to hijack common iron trafficking pathways to enter cancer cells. However, unlike iron, gallium cannot take part in cellular redox reactions, thus disrupting critical iron-dependent processes and resulting in cell death. Our group has demonstrated the effectiveness of gallium maltolate (GaM), a newer generation compound with high oral bioavailability and therapeutic index, both in vitro and in vivo. In animal studies of adult glioblastoma, the most aggressive type of primary brain tumors, we demonstrated a slower tumor growth rate, a doubling of survival, and an improved quality of life with treatment. Preliminary studies in pHGGs and ATRTs suggest similar benefits. Despite this promising initial step, questions regarding the efficacy of GaM remain. Specifically, we intend to address the knowledge gap as to what drives response to GaM therapy. Therefore, our overall goal is to be able to offer a new treatment strategy to brain tumor patients with few therapeutic options. In Specific Aim 1, we propose a sophisticated multi-pronged approach leveraging behavioral assessments, state-of-the-art MRI-guided biopsy, cellular bioenergetics, and induction coupled plasma mass spectrometry (ICP-MS), to develop a tissue sensitivity profile. In Specific Aim 2, we propose to explore the synergistic potential of combining GaM with radiation therapy, the backbone of many treatment protocols. Preliminary data suggest a potentiating cytotoxic effect, which requires confirmation in vivo. Collectively, these investigations will provide insights regarding how to maximize antineoplastic therapy with GaM to improve both the quality of life and extend survival in children with primary brain tumors.

Garfield Weston Foundation

One of the UK's largest family-funded charitable foundations distributing 90M+ GBP annually across arts, education, community, environment, health.

Garinagu Houngua Planning Committee

Garinagu Houngua Planning Committee will organize a series of workshops for community members to learn about various aspects of Garifuna culture. The series will take place at Casa Latina located at 317 17th Ave S. Workshops will be held from 3 pm - 5 pm each Saturday and Sunday beginning September 28, 2019 and ending April 11, 2020.

NIGMS - National Institute of General Medical Sciences

ABSTRACT Members of the voltage-gated superfamily of tetrameric cation channels have central roles in human physiology. My group's long-term objective is to understand the molecular mechanisms of gating, lipid modulation and cellular function of two families of channels within this group: the large conductance and calcium activated potassium (BK) channels and the cyclic nucleotide-modulated (CNM) channels, which have been the topics of 2 distinct RO1s from NIGMS (GM088352 – K channels, and GM124451 – CNM channels). BK channels have the ability to couple intracellular Ca2+ to membrane potential variations, play major physiological roles in vascular smooth muscle tone maintenance, regulation of circadian rhythms, hearing, neurotransmitter release. CNM channels are activated by cyclic nucleotides (CNG), and hyperpolarization (HCN), and are expressed in the heart and brain where they play key roles in pacemaking, vision, olfaction. They are excellent drug targets and understanding how they gate can be therapeutically useful. BK channels can associate with tissue-specific accessory subunits, endowing the channels with different functional properties. 2 and 3 subunits induce N-type (or ball-and-chain) inactivation of the otherwise non-inactivating BK channels. The structural correlates of this process were not known. We previously determined the structural correlates of ball-and-chain inactivation in MthK, a prokaryotic homolog of BK channels from Methanotropicum thermoautotrophicum, and found it has a strong lipid dependence. We propose to determine the structural correlates of ball-and-chain inactivation in BK channels and understand how lipids modulate inactivation in both BK and MthK channels. We previously used SthK, prokaryotic homolog of CNM channels from Spirochaeta thermophila, as model to investigate CNM channel gating. We determined the mechanisms of ligand selectivity and increase in activity with anionic lipids in SthK, and our preliminary data shows that lipids modulate the temperature dependence of SthK. We propose to elucidate the molecular mechanism of this process and determine whether it is conserved in eukaryotic thermo-sensitive channels. In addition, we propose to elucidate the structure of the olfactory CNG channel, proposed to be a heteromer of CNGA2, CNGA4, and CNGB subunits and determine the mechanism of lipid modulation and ligand selectivity. Since membrane lipid composition cannot be controlled in cells, we will use a bottom-up approach of purified channels in reconstituted systems, to rigorously control lipid content. We combine state-of-the-art techniques: single-particle cryo-EM, atomic force microscopy, computational approaches, and functional assays to reach our goals. The accomplishment of these projects will provide a comprehensive picture of ball- and-chain inactivation in calcium-gated potassium channels and its lipid dependence, as well as of ligand selectivity, lipid modulation and temperature dependence in select CNM channels.

NIA - National Institute on Aging

PROJECT SUMMARY Microglia are the macrophages of the brain and become activated in response to amyloid. Recently, single cell sequencing has defined multiple activated states of microglia including two states that are robustly induced in animals of Alzheimer’s disease (AD): disease associated microglia (DAM), and interferon responding microglia (IRM). It is now established that in response to amyloid microglia initiate the classical pathway of the complement cascade, and that the complement cascade is a critical mediator of neuronal synapse loss during disease progression. Synapse loss is among the strongest neurobiological correlates of cognitive decline in AD. Global ablation of the C1 complex (via C1qa gene knockout) preserves synapses in AD mouse models, highlighting the importance of determining the mechanisms determining the role of microglia in complement-mediated synapse loss. Yet despite much work, key knowledge gaps remain. First, the relationships among the different transcriptionally defined microglia states have not been determined. Second, all microglia express C1Q and it remains unknown whether microglia belonging to distinct states trigger synapse loss on neurons. Third, the complement cascade requires downstream components such as complement factors C2 through C9 that are not expressed by microglia, but virtually nothing is known about the spatial and temporal coordination of the specific cell types expressing these components in the brain. Filling these knowledge gaps may lead to new therapeutic avenues that prevent or intervene in synapse loss in AD. By leveraging floxed alleles of Csf1r, Trem2, Sting1, C1qa, C3, C5 and C7, microglia state specific Cre driver lines such as Cx3cr1-cre, Tmem119-cre, Itgax-cre, and Mx1-cre, and reporter lines to lineage trace distinct states, we will take a multi-modal approach based on genetic strategies to address these questions with cellular specificity. We will use distinct mouse genetic contexts we have shown are susceptible (C57BL/6J) or resilient (PWK/PhJ) to synapse loss, and we will employ state-of-the- art methodologies including single cell myeloid cell sequencing, spatial transcriptomics and protein visualization, and circuit-specific labeling of synapses. In three aims we will test the model that IRM are an intermediate microglia state necessary to recruit DAM to plaques, and that DAM are the critical state driving complement- mediated synapse loss. In Aim 1, to test whether IRM are the intermediate state between homeostatic microglia and DAM, we will lineage trace IRM, ablate DAM or IRM, and conditionally delete Sting1 (a key mediator of interferon signaling) from DAM. In Aim 2, to determine whether DAM are the primary initiators of complement mediated synapse loss, we will conditionally delete Trem2 from homeostatic microglia, ablate DAM, and conditionally delete C1qa from DAM. In Aim 3, to uncover the cell types producing the downstream components of the complement cascade, we will perform spatial transcriptomics and protein visualization. We will then conditionally delete a downstream component from its parent cell type. Successful completion of these aims will result in the identification of critical cellular and genetic contributors to complement-mediate synapse loss in AD.

NIDA - National Institute on Drug Abuse

PROJECT SUMMARY Early initiation of cannabis use (<16 years of age) increases risk for cannabis use disorder (CUD), mental illness, cognitive impairment, later unemployment, and poor social relationships. Prevention of early initiation is critical for improving social and health outcomes. Precision prevention programs have reduced youth substance use, but no approaches have specifically targeted cannabis use. Furthermore, no studies have comprehensively considered risk factors for early cannabis initiation (genetic, social, behavioral, environmental, and cognitive) to enhance the prediction of early use and inform precision prevention approaches. Comprehensive multivariable prediction models for early cannabis initiation that include genetics and social/environmental factors are needed. Cannabis use is polygenic, influenced by multiple genetic variants with weak-to-moderate effects, and polygenicity makes it difficult to translate genetics for clinical application. One method for clinically applying genetics is through the development of polygenic risk scores (PRS) that are composite scores representative of overall genetic risk. Prior PRS have typically lacked portability to non-European populations; however, a state- of-the-art method has been developed to build PRS with significantly improved risk prediction (34% improvement) across ancestries. There is a need to apply this method to develop cross-ancestry PRS for cannabis use for inclusion of overall genetic risk in comprehensive prediction models. Furthermore, given the complex interplay between genetics and social/environmental factors, research is needed to understand gene by environment (GxE) interactions in which social/environmental factors synergistically impact the risk conferred by genetics. Research into GxE interactions is statistically and computationally challenging, and traditional single-variant and more recent polygenic approaches focus on lower order 2-way interactions. Our logic forest (LF) algorithm efficiently searches all possible interactions up to 8 variables without a priori specification. This study will apply these state-of-the-art computational methods to the Adolescent Brain Cognitive Development (ABCD) Study, which examines childhood risk factors and initiation of substance use from ages 9-10 years to early adulthood in a population demographically reflective of the U.S. Nearly all youth had not used cannabis at recruitment, enabling the prospective measurement of initiation and the development of prediction models integrating genetics with pre-substance use measurements of cognitive, social, and environmental factors. This research will 1) develop cross-ancestry PRS for inclusion in prediction models that comprehensively consider genetic, sociodemographic, behavioral, cognitive, and environmental factors, and 2) apply LF to gene-sets within known biological pathways across the whole genome to identify pathway-specific GxE interactions. Comprehensive models coupled with a more complete understanding of GxE factors influencing early cannabis initiation can identify 1) high risk youth populations for targeted prevention, 2) targetable factors present among high-risk clusters for tailored interventions, and 3) biological pathways for therapeutic development.

NIAID - National Institute of Allergy and Infectious Diseases

ABSTRACT The goal of the studies in this proposal is to establish how disease-associated genetic variation regulates antibody responses at the level of B cell help by follicular helper T cells (TFH). In Aim 1, we will use a powerful combination of state-of-the-art, variant-to-gene mapping approaches in follicular T cells ‘caught in the act’ of helping B cell antibody responses in human lymphoid tissue: 1) chromosome conformation data to reveal physical associations between regulatory variants and gene promoters in the context of the 3D structure of the genome, 2) quantitative expression-trait mapping to reveal statistical associations between disease-associated variants and gene expression in TFH cells, 3) massively-parallel reporter assays to identify expression- modulating variants, and 4) CRISPR interference screens to identify genes regulated by antibody disease variants in TFH cells. In Aim 2, ‘novel’ effector genes prioritized by the approaches in Aim 1 will be assessed for roles in TFH function in an in vitro lymphoid organoid model human humoral immunity and in in vivo mouse models of lupus and influenza vaccination. The proposed studies are supported by the extensive experience and preliminary data of the research team, and will use a confluence of evidence from orthogonal approaches to power discovery of novel mechanisms that regulate T cell-dependent humoral immunity, refine our understanding of how common genetic variation contributes to autoimmune disease susceptibility, and point to novel therapeutic interventions to modulate humoral immunity in the context of vaccines, infectious disease, and autoimmune disorders.

NHLBI - National Heart Lung and Blood Institute

PROJECT SUMMARY Gene editing offers the prospect of directly modifying any nucleotide(s) in the genome, including correction of pathogenic variants underlying disease. These technologies could form the basis of cures for currently untreatable genetic conditions. The work proposed in this application aims to identify genome editing strategies to prevent aortic pathology in Marfan syndrome (MFS). MFS is the most prevalent hereditary connective tissue disorder and is associated with significantly increased morbidity and mortality due to life-limiting thoracic aortic aneurysm and dissection. MFS is caused by heterozygous pathogenic variants in FBN1, the gene encoding the main structural component of extracellular microfibrils, fibrillin-1. Microfibrils are essential for providing structural elasticity and resilience, in addition to having a signaling role, and defects in both features are thought to contribute to elastic lamina fragmentation and aortic wall weakness. We hypothesize that gene editing correction of FBN1 pathogenic variants or genome editing-based upregulation of the structurally related protein FBN2 within aortic vascular smooth muscle will reduce risk of aortic root dilation and dissection, thereby limiting the major cause of morbidity and mortality in this disease. In the first aim, the candidate will identify a prime editing strategy to correct the Fbn1 C1041G pathogenic variant in aortic vascular smooth muscle of a murine model of MFS and will test this therapeutic strategy by monitoring aortic aneurysm. In the second aim, a machine learning model will be used to identify promoter variants, putative enhancers, and transcription factor binding site motifs within the FBN2 promoter region that are predicted to augment FBN2 expression. These elements will be functionally evaluated in a massively parallel reporter assay (MPRA). Finally, in vivo genome editing will be used to introduce an optimized FBN2 upregulatory strategy established through these analyses in the Fbn1C1041G/+ mouse model of MFS for correction of aortic pathology. In addition to establishing potential gene editing treatment strategies for aortic aneurysm in Marfan syndrome, these studies will enable the candidate to obtain expertise in the design and application of state-of-the-art gene editing tools that can be used to generate disease models and investigate therapies for many different genetic disorders, which he plans to pursue throughout his career as a physician-scientist.

NHGRI - National Human Genome Research Institute

Project Summary This proposed MAGen development site aims to develop genomics and multi-modal artificial intelligence (AI) models to transform personalized cancer risk assessment, monitoring, and prevention. A substantial gap exists between the theoretical potential of genomics-based AI predictions and their practical application in clinical and population healthcare settings. The clinical classification of genetic variants is hindered by insufficient data to classify ultra-rare variants, particularly those found in non-European populations. Moreover, despite significant advances in AI across fields, we lack AI models that can combine diverse streams of health data to accurately predict disease risk across a person’s life course. Finally, the real-world effectiveness of these AI models remains untested and their ethical, legal, and social implications (ELSI) are unclear. To address these challenges, our primary goal is to develop state-of-the-art (SOTA) AI models that can accurately identify pathogenic variants affecting DNA repair genes and predict cancer risks over the life course of high-risk carriers, thereby optimizing screening and prevention strategies in an ELSI-informed manner. Our multidisciplinary team comprises experts in computational genomics, AI/ML, health informatics, statistical genetics, medical genetics, population health, oncology, and ELSI research from Icahn School of Medicine at Mount Sinai (ISMMS), Boston Children’s Hospital/Harvard, and Columbia University, and has complementary and extensive experience in consortium and team science projects. In our proposed project for MAGen, Aim 1 will develop robust genomic AI models for identifying protein-disrupting missense variants that confer high cancer risks. Aim 2 will combine other genetic factors, including common and rare variant polygenic risk scores (PRS), and non-genetic factors, including EHR, longitudinal lab markers, SDoH, and digital pathology, to predict cancer risk over the life course and optimize screening recommendations for carriers. Aim 3 will cross-validate AI models in real-world population biobanks and determine their clinical impact. Aim 4 will construct an ELSI framework and conduct ELSI projects to evaluate the multi-faceted impacts of AI-driven genetic diagnostics. Aim 5 will disseminate AI model/predictions, cross-validation data, and ELSI recommendations. The completion of these Aims will bring genomics-based and multi-modal AI closer to the advancement of personalized medicine in real-world settings by more accurately classifying pathogenic variants, optimizing the timing of screening, and identifying key lifestyle and medical prevention strategies that could ultimately save lives from cancer.

Georgetown Merchants Association

The Georgetown Carnival is a free, one-day, arts & music-focused street fair centered in Georgetown’s historic business district. The 2025 Carnival will take place on June 14, 12-10 pm, on Airport Way S between 12th Ave S & Corson Ave. The Carnival is a celebration of the neighborhood and its unique place as an artistic and small business haven surrounded by large-scale industry. It brings visibility to the neighborhood artists and businesses, and gives our small residential community a day of fun focused on our special neighborhood. With a residential population of only 1700, Georgetown’s small businesses rely on customers from other parts of Seattle to remain viable. Events like Carnival are vital in increasing the visibility of our local artists and businesses, and create a lasting economic impact by introducing new people to the neighborhood and encouraging them to return. In 2023, we welcomed over 5,000 people to the Carnival, many of whom were first time visitors to Georgetown.