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The Magnet School for Technology and Communication

Conversion of (1) classroom into a news network consisting of a sound proof roomallowing editors to monitor the visual and audio displayed and heard on the monitors.

Somali Health Board

This is annual health fair that promotes well-being of the community.Health systems will come into the community and provide health screening, education,where their patients reside, at their own comfort.This fair requires a lot of planning & as such we have dedicated a health fair team to coordinate this. We'll be designating volunteers & various committees to ensure a successful fair.Funds from this grant will help us expand the # of residents reached and services provided this year.

DEPT OF DEFENSE.DEPT OF THE NAVY.NAVSUP.NAVSUP WEAPON SYSTEMS SUPPORT.NAVSUP WSS MECHANICSBURG.NAVSUP WEAPON SYSTEMS SUPPORT MECH

10--CAMO CVR LWR BARREL

East African Community Services

Expand the Science, Technology, Engineering and Math (STEM) enrichment program to engage and empower 40 East African girls with strong critical thinking and digital literacy skills so they are ready to pursue opportunities in higher education and jobs in high-demand technical fields.

NEI - National Eye Institute

Project Summary Cephalopod neuroscience offers a unique perspective in comparative brain research. Similar to vertebrates, cephalopods have evolved large, complex brains, enabling remarkable sensory, motor, and cognitive abilities. They exhibit sophisticated behaviors such as independent control of eight flexible arms, dynamic skin patterning for camouflage, and advanced learning and decision-making capabilities. Understanding the cephalopod nervous system has the potential to uncover fundamental principles of brain organization and function across species. Despite their fascinating neurobiology, the mechanistic workings of cephalopod brains remain largely unexplored. However, recent technological advances have catalyzed rapid progress and an influx of new researchers into the field, leading to the establishment of the first Cephalopod Neuroscience Gordon Research Conference. This meeting will bring together scientists from diverse areas of cephalopod research, including genomics, neural development, systems neuroscience, computation, and tool development. Our key objectives are to: (1) foster knowledge exchange and highlight recent discoveries, (2) cultivate an engaged and collaborative research community, and (3) facilitate resource and technique sharing. A strong emphasis will be placed on supporting trainees to ensure broad participation, and provide a strong foundation for this new Gordon Conference in the future. By combining cutting-edge science with community-building efforts, this conference aims to accelerate advances in cephalopod neuroscience and provide insight into broad principles of brain function.

NHLBI - National Heart Lung and Blood Institute

PROJECT SUMMARY The objectives of the 2026 GRC “Lymphatics” are to present state-of-the-art research on the lymphatic vascular system, to fuel scientific exchange across scientific disciplines and expertise, and to spearhead new ideas and collaborations. The meeting will bring together investigators from all backgrounds and from all career stages, converging on lymphatics as one common theme. Specific Aims are (1) To present conceptual advances in research of the lymphatic vascular system and its pathologies; (2) To discuss paradigm shifting findings of lymphatics in different organ systems and their contribution to disease; (3) To integrate knowledge from different disciplines; and (4) To nurture junior investigators and trainees entering the lymphatic research field. Sessions will cover conceptual advances in development and disease of lymphatic vessels from basic aspects to potential clinical applications. The meeting will highlight recent insights into the diversity and organ-specificity of lymphatic endothelium. Novel molecular regulators for lymphatic vasculature during bone development, Schlemm’s canal in the eye and lacteals in the intestine will be discussed. New factors regulating lymphangiogenesis and lymphatic integrity will be introduced. Advances in research on lymphatic malformations, lymphedema and other congenital lymphatic diseases will be discussed. The meeting will also present emerging concepts on the role of lymphatics in atherosclerosis, myocardial infarction, and congestive heart failure. Paradigm shifts in the field will be presented, and crosstalk between lymphatics and immune system discussed. Advances in understanding how lymphatics contribute to disease, including cardiovascular inflammation, cancer, autoimmune and neurodegenerative diseases will be highlighted. A key goal for this meeting is to serve as a platform for growth of lymphatic research, and for helping to promote new scientists entering this research field. Abundant opportunities for the presentation of research by early career investigators will be provided. The GRS on Lymphatics organized by junior investigators will provide opportunities exclusively for junior investigators to present their work and form collaborations. Aside from presentation and poster sessions, a career development session will also be held during GRS to provide guidance to those just getting their start in the lymphatic research field, and to discuss strategies for how to navigate issues impeding lymphatic research or obstacles to participation by incoming lymphatic researchers. A patient session will also be held to allow researchers to hear from those suffering from lymphatic diseases and hear their perspective on the potential promise of translating lymphatic research into new therapies. The sponsoring organization is dedicated to advancing the frontiers of science. In accordance with its mission, this conference will place emphasis on the presentation of unpublished data, high-quality science and rich discussions in many fields, such as cardiovascular disease, cancer, immunology, metabolism and neurobiology. We anticipate that the opportunities provided to young investigators participating in this conference will facilitate their rise as future leaders of the lymphatic research field.

NCCIH - National Center for Complementary and Integrative Health

Project Summary Natural products research involves highly interdisciplinary fields focused on translation of chemically and biologically complex natural products to applications for human health and health outcomes. The evolution of natural products as potentially multifunctional ligands with highly specific affinities for health-relevant targets has provided opportunities to develop therapeutic agents for cancers, infectious diseases, and neurological disorders, as well as for multisystem, multicomponent diseases and conditions. Natural products from the marine environment have provided unique molecular scaffolds that interact with biological targets and signaling pathways to effect changes in phenotype and physiology, resulting in several that have translated to clinical use. In addition, transformational advances in biosynthesis, target profiling, and analytical technologies are poised for incorporation in computational tools using machine learning for discovery of natural products and their biological signatures. The 2026 Marine Natural Products (MNP) Gordon Research Seminar (GRS) and Gordon Research Conference (GRC) are planned for February 28 – March 6, 2026, in Ventura, California. The GRS is a unique forum for graduate students and postdoctoral fellows to present and exchange new data and innovative ideas across the scope of basic and applied MNP research. The GRC will bring together early career and established researchers from academia, industry, and government (including all GRS participants) to address the discovery, characterization and functional development of MNPs. Sessions will focus on technological advances to spur new discovery, discovery of new molecular targets and receptors, computational informatics challenges and opportunities, symbiosis and microbiome interactions to facilitate discovery, biosynthesis and bioengineering for molecule and target discovery, bioactive molecules from marine sources, integrating novel synthetic approaches for enhanced discovery, and marine biogeochemistry to address microbial challenges. Our specific aims for the conference include: (1) providing a forum for emerging and established scientific leaders to present cutting-edge MNP research; (2) stimulating multi-disciplinary collaborations and providing opportunities for interdisciplinary activities; (3) highlighting biological targets, physiological function, and translational applications of MNPs; and (4) providing training for graduate students and early career scientists to promote broad participation. The collegial atmosphere of the MNP GRS and GRC will foster collaborations, provide training opportunities, and promote exchange of ideas amongst leaders in the field and new participants. NIH support will be used to enhance participation by graduate students, postdoctoral fellows, and early career faculty at the GRC, as well as for postdoctoral fellows and graduate students attending the GRS. Ample opportunities for scientific exchange and networking, both in and outside of meeting sessions, will foster new collaborations that bridge scientific disciplines and potentiate paradigm shifts in natural product sciences.

NINDS - National Institute of Neurological Disorders and Stroke

Project Summary Building upon the success of past meetings, the 2026 Gordon Research Conference (GRC) on Thalamocortical Interactions will bring together leading neuroscientists, from early career to established investigators, to discuss the latest breakthroughs in understanding the functions of the nervous system influenced by the thalamus. The scientific sessions will highlight cutting-edge discoveries on thalamus-brain interactions in normal and disease conditions. Session topics will include the role of the thalamus in brain plasticity and stroke recovery, movement, decision-making, development, and visual and tactile sensory processing, as well as topics new to this GRC, such as thalamic contributions to pain and itch processing, conscious states, addiction, and motivation. A major goal of this international conference is to foster interactions among scientists at all career stages, from graduate student and postdoctoral fellows to early career and senior investigators and spanning wide-ranging areas of expertise and interests to brainstorm new hypotheses related to thalamic function and to stimulate new collaborations. Leading researchers in the field will present their most current, unpublished work with extensive discussion planned for each session. Discussion will be further fostered during time set aside for more informal interactions. To promote the next generation of neuroscientists, short talks will be selected from abstracts submitted by graduate students and postdoctoral fellows. In addition, junior faculty represent a third of confirmed invited speakers. Finally, this GRC will be preceded by a Gordon Research Seminar (GRS), organized by graduate students and postdoctoral fellows, which will feature talks and posters by trainees. The GRS will serve as a forum for trainees to present their work and to broaden their peer network and will complement the GRC in bringing together scientists from around the world to discuss and identify new directions in thalamus-brain interactions. The 2026 conference promises to continue the highly regarded tradition and reputation established by the decade-long history of the Thalamocortical Interactions GRC series, bringing top neuroscientists from around the world to communicate innovative science, develop new hypotheses of thalamus function, establish new collaborations among participants, and nurture the next generation of young neuroscientists.

Maria Teresa Mirabal Middle School

Purchase 50 ActivTables (1 for every classroom). 10- Ultimate Polycarbonate Basketball System, (2) Pro premier Soccer Goal 8'Hx24'Wx3'Dx9', (10) Soccer Flags, (30)Mis-Size Laptop School Image Lenovo L430 for Classroom Use, (150 all in Ine Space Saving PC School Image Lenovo.

DEPT OF DEFENSE.DEFENSE LOGISTICS AGENCY.DLA LAND.DLA LAND COLUMBUS.DLA LAND AND MARITIME

28--GUIDE,VALVE STEM

DEPT OF DEFENSE.DEPT OF THE NAVY.NAVSUP.NAVSUP WEAPON SYSTEMS SUPPORT.NAVSUP WSS MECHANICSBURG.NAVSUP WEAPON SYSTEMS SUPPORT MECH

30--CLUTCH ASSEMBLY,FRI, IN REPAIR/MODIFICATION OF

NHGRI - National Human Genome Research Institute

Project Summary/Abstract This proposal will investigate the dynamic 3D folding of the genome at ultra-high resolution across length and time scales as it relates to gene activity. Studies in Aim 1 will investigate how newly discovered “microcompartments” between active cis-regulatory elements (CREs) form as cells exit mitosis and enter G1 phase. Because microcompartments cannot be resolved with conventional Hi-C or Micro-C methods, we will apply ultra-high resolution Region Capture Micro-C (RCMC). Using acute degradation technology, we will investigate the role of a battery of key proteins in microcompartment formation. Studies at the mitosis-to-G1 transition will be complemented using a dynamic cell transition system that considers CRE dynamics during transcriptional repression. Genome-wide insights will be obtained using newly developed machine learning- based imputation. Aim 2 is motivated by the hypothesis that similar to transcriptionally active microcompartments, intricate fine scale chromatin organization exists within heterochromatin. We will apply RCMC to representative regions of constitutive and facultative heterochromatin. These studies will be complemented by experiments perturbing key heterochromatic regulators. Furthermore, machine learning based imputation will be applied to obtain genome-wide insights, followed by validation experiments, aimed at uncovering the fine-scale microstructure of the repressive chromatin compartments. Importantly, both aims will be enhanced by mechanistic 3D polymer modeling with the goal of developing a polymer model from first principles that can explain the data, to make experimentally testable predictions, and to estimate key parameters which are otherwise not experimentally observable. In Aim 3 we will validate and extend our findings using super-resolution chromosome tracing experiments as an orthogonal and sequencing independent method. This will address if microcompartments form through simultaneous multi-way interactions, will reveal cellular or allelic heterogeneity in chromosomal folding, and deepen our understanding of the relationship between 3D genome folding and transcription.

Xbot Robotics

Provide Science, Technology, Engineering and Math (STEM) training in 3D modeling and 3D printing to middle and high school girls and educators in underrepresented communities

DEPT OF DEFENSE.DEPT OF THE NAVY.NAVSUP.NAVSUP WEAPON SYSTEMS SUPPORT.NAVSUP WSS PHILADELPHIA.NAVSUP WEAPON SYSTEMS SUPPORT

53--BOLT

DEPT OF DEFENSE.DEPT OF THE NAVY.NAVSUP.NAVSUP WEAPON SYSTEMS SUPPORT.NAVSUP WSS MECHANICSBURG.NAVSUP WEAPON SYSTEMS SUPPORT MECH

61--RUPTURE PANEL

DEPT OF DEFENSE.DEPT OF THE NAVY.NAVSUP.NAVSUP WEAPON SYSTEMS SUPPORT.NAVSUP WSS MECHANICSBURG.NAVSUP WEAPON SYSTEMS SUPPORT MECH

70--COMPUTER,DIGITAL, IN REPAIR/MODIFICATION OF

Sage Library System

The Sage Library System covers literally half of the state of Oregon, including over 70 libraries of various types. Unfortunately, the varying skills levels in the system, paired with recent growth in the consortium, has resulted in an inconsistent bibliographic database. This frustrates patrons who use the online catalog (http://catalog.sage.eou.edu) and makes it difficult for them to find what they want. Sage seeks funds to hire a fulltime Cataloging Specialist to train Sage members in cataloging.  They will also clear up database inconsistencies, and lay the groundwork for consistent cataloging so that Sage can better connect its patrons with the items they want.

Baker County Library District

The Sage Library System covers half of the state of Oregon, including over 70 libraries of various types. Unfortunately, the varying skill levels in the system paired with growth in the consortium have resulted in an inconsistent bibliographic database. This frustrates patrons who use our online catalog and makes it difficult for them to find what they want. Sage hired a full-time Cataloging Specialist to train Sage members in cataloging, clear up database inconsistencies, and lay the groundwork for consistent cataloging so that Sage can better connect its patrons with the items they want.

NIAID - National Institute of Allergy and Infectious Diseases

Infants are especially susceptible to intracellular pathogens because their immune system is not fully equipped to fight against these microorganisms. As a result, infant infections are a leading cause of mortality, with >1.5 million children dying of infections before 5 years of age in 2022 alone. The current consensus is that before birth, conventional T cells are skewed in favor of T regulatory or T helper (Th) 2 responses, leaving neonates and infants more vulnerable to pathogens cleared by Th1 immunity. Therefore, Vγ9Vδ2 (or simply Vδ 2) T cells, are particularly important in early life, because they are poised to secrete Th1 cytokines even before birth and acquire potent cytotoxic function shortly after birth. Despite their protective role against pathogens, human Vδ2 cells, which are absent in mice, are not well studied in neonates and infants. Our long-term goal is to elucidate their functional heterogeneity because understanding their features in the first few months of life will allow us to harness their properties to protect infants from infections. This task is challenging for many reasons, including the difficulty in obtaining samples from the infants at highest risk of early infections, such as premature babies. Our recent observations, obtained using spectral flow cytometry (SFC) and single cell RNA-seq, converge to show phenotypic and functional heterogeneity of cord blood (CB) Vδ2 cells, with heightened stemness compared to their adult counterpart and a cluster of PD1-hi cells (absent in adults) that may follow a distinct functional program specific for the early life stage. We posit that differences in Vδ2 cell cluster composition at birth have functional consequences and result in improved or decreased antimicrobial activity depending on the composition. The resulting overarching hypothesis is that human neonatal V δ2 lymphocytes exist in heterogenous functional/differentiation states impacting host immune competence in the critical early life window. As a multidisciplinary team of immunologists and computational biologists, we propose to characterize cord blood Vδ2 cells in existing specimens, including samples of premature babies, using state of the art techniques –Cellular Indexing of Transcriptomes and Epitopes by sequencing (CITE- seq) and SFC- with the following aims. Aim 1. Evaluate the phenotypic and functional heterogeneity of Vδ2 T cells at birth and in early life in relation to age, with a focus on a unique subset of PD1-hi cells present in cord blood. Aim 2: Assess how the composition of Vδ2 T cells at birth impacts function of these cells at the population level, combining CITE-seq analysis with depletion and transduction experiments. The goal of this proposal is to develop a robust molecular and functional map of a critical subset of innate-like T cells, which will provide a valuable reference for the cellular heterogeneity in the neonatal human immune system and a foundation for future proposals aimed at understanding Vδ2 cell biology in early life.

NSF

Researchers who study human diseases or test new drugs often use microfluidic devices that contain embedded cells that mimic the behavior of specific organs. The usual approach is to make a change in the cells’ environment and observe changes in the health of the cells. This project will expand that approach by finding ways to control the health of the cells as their environment changes. The project will create an “organ-on-a-controller” system that controls the health and function of human liver cells called hepatocytes. The system will integrate three components: 1) miniature sensors that monitor multiple vital signs of the hepatocytes in real-time, such as protein production and metabolite levels; 2) a computer model that learns how the cells respond to different drugs or nutrients; and 3) an intelligent control system that uses this knowledge to automatically adjust the input to the cells so that a particular cellular health state and function can be achieved. This approach will keep cells healthy and will guide unhealthy cells from a diseased state, such as fatty liver disease, back toward a healthy one. The technology will create a powerful tool that can accelerate the discovery of safer and more effective drugs, advance personalized medicine, reduce the need for animal testing, and provide a deeper understanding of complex chronic diseases. Results will help advance new concepts in biotechnology and advanced biomanufacturing. A fundamental gap exists in our ability to dynamically control complex biological systems. Current in vitro microphysiological systems (“organs-on-chips”) are largely open-loop, precluding the active regulation of cellular function based on real-time feedback. This project aims to address this knowledge gap by creating a first-of-its-kind “organ-on-a-controller” platform that integrates multiplexed biosensing, predictive modeling, and adaptive closed-loop control to actively steer cellular function. Using primary human hepatocytes as a biologically relevant model system, this project will design an integrated microfluidic platform for the simultaneous, real-time measurement of key secreted factors and intracellular reporters of transcription factor activity. Our approach will provide a continuous, multi-parameter view of the cellular state with high temporal accuracy. Further, a library of predictive mathematical models (transfer functions) will be developed that describe the dynamic input-output relationships of hepatocytes in response to metabolic and inflammatory stimuli. A sophisticated model predictive control will be implemented and validated to actively maintain hepatocyte homeostasis under inflammatory challenge and steer cells from a disease state toward a healthy phenotype. By closing the loop between sensing and actuation, the platform will be inherently adaptive, learning from cellular responses to account for biological variability and perturbation. 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.

NIA - National Institute on Aging

Summary/Abstract Vascular pathology has been identified as a critical driver of Alzheimer’s disease. This proposal aims to establish a human induced pluripotent stem cell (iPSC)-derived model to study the impact of genetic and environmental factors on the development of Alzheimer’s disease-related vascular pathology. Most Alzheimer’s disease patients have cerebral amyloid angiopathy (CAA), the symptoms of which include build-up of amyloid around vessels, cerebral hemorrhages, microbleeds, and inflammation. Studies in multiple mouse models of Alzheimer’s disease have demonstrated that hemopoietic-derived brain-resident microglia and circulating monocytes and perivascular macrophages can be protective against CAA. The APOE44 genotype is the strongest genetic risk factor for Alzheimer’s disease and CAA. Despite the clear interplay between Alzheimer’s disease, CAA, and APOE genotype, the molecular mechanisms underlying the vascular changes are not fully understood. These observations motivate us to create a model using human cells to study the underlying mechanisms of microglia- vascular interactions in cells of different APOE genotypes. In prior published studies, we created a 3D human iPSC-derived vascular model in a hydrogel scaffold. It undergoes vasculogenic and angiogenic events and forms a model plexus (the VAMP model). We showed that the VAMP model can be built inside a microfluidic device to enable regulatable perfusion, mimicking blood flow. We have advanced the model by incorporating microglia (VAMP-MG) to reflect the key cell-cell interactions in CAA better. The work proposed in two specific aims will increase the utility of the VAMP-MG model in five significant ways: We will 1) build the model from a recently established collection of APOE44 versus APOE33 isogenic iPSC lines to incorporate genetic risk factors; 2) incorporate live reporters to monitor the activation states of microglia and vascular cells in real-time; 3) create the VAMP-MG model in microfluidic chips to achieve perfusable vasculature; 4) flow human plasma from Alzheimer’s disease, aged-matched healthy, or young healthy donors through the vessels to improve modeling of disease-relevant environmental factors; 5) use Ribo-Tag technology to identify transcriptomic changes in the vascular endothelial cells and the microglia. The model will be deeply characterized and validated at cellular and molecular levels. Alzheimer’s disease-relevant phenotypes, including amyloid deposition and clearance, A secretion, and inflammatory factor production, will be assessed. We will compare transcriptomic data collected from the model to published human Alzheimer’s disease brain versus healthy control brain transcriptomic data, including single nuclear RNA-seq datasets. Completing the proposed work will enhance our ability to study vascular and microglial responses to inflammatory signals in real time, generate transcriptomic data at cell type levels, and establish APOE44 versus APOE33 phenotypes in microglia-vascular models to a new level of complexity. In the future, this VAMP-MG model can potentially define key genetic and environmental factors that exacerbate or alleviate Alzheimer’s disease-vascular phenotypes.

NIAID - National Institute of Allergy and Infectious Diseases

SUMMARY Regulatory T cells (Tregs) dominantly suppress effector T cells, serving as a living therapeutic agent for immunological diseases such as type I diabetes and multiple sclerosis. However, Tregs exhibit a plastic fate at inflammatory conditions characterized by the loss of lineage identity and the acquisition of effector function, which compromises their efficacy. Furthermore, human Tregs cannot be reliably sorted at high purity by surface markers, because conventional T cells transiently upregulate Foxp3 expression upon activation. The presence of contaminating effector T cells would exacerbate inflammation. Technically, retroviral or lentiviral systems, which are widely employed for engineering Tregs, are susceptible to integration site-related effects known as position effects, resulting in substantial variations in gene expression levels. These factors pose challenges for modifying Tregs with enhanced performance for research and therapeutic applications. Therefore, expressing antigen-specific receptors or booster genes in Treg samples presents significant uncertainties. To address this issue, we hypothesize that tethering the Treg master regulator Foxp3 to genes of interest will generate Tregs with high lineage fidelity and robust regulatory function. In preliminary experiments, we began testing this hypothesis. We creatively combined CRISPR/Cas9 genome editing with bacteriophage integration system to insert genes of interest into the 3’ untranslated region (3’-UTR) of the endogenous Foxp3 gene. We leveraged CRISPR/Cas9 genome editing to first insert a landing pad attP site at Foxp3 3’-UTR and subsequently performed site-specific insertion of genes of interest via the attP-attB integration system. We validated this strat- egy in experimental mice and generated knock-in strains bearing Tregs with three representative features, demonstrating the feasibility of our approach. Our method surpasses ectopic Foxp3 expression via retroviral or lentiviral systems in preventing Treg fate loss. The latter cannot convert contaminating conventional T cells into fully functional Tregs, and Foxp3 expression is significantly influenced by the insertion sites. Based on preliminary results, we propose to rigorously test our hypothesis. We will establish a versatile platform to engineer Tregs by expressing genes of interest under the control of endogenous Foxp3 gene via its 5’-UTR or 3’-UTR. This will be first tested in murine Tregs through genome editing in germline. Subsequently, we will extend our approach to human primary Tregs and develop protocols for inserting genes into Foxp3 5’- UTR or 3’-UTR via sequential CRISPR/Cas9- and integrase-mediated insertions. Successful completion of our study will establish an innovative method to engineer Tregs with high lineage fidelity and robust expression of modifying genes, thereby minimizing the consequences of transdifferentiation of Tregs into disease-causing ef- fector T cells or of empowering contaminating conventional T cells. Tregs engineered through this approach will serve as a reliable source for both basic research and translational applications.

NIA - National Institute on Aging

Project Summary/Abstract Epidemiological data on Alzheimer’s disease (AD) paint a bleak picture for American society, with the death toll from the disease surpassing that from breast cancer and prostate cancer combined. As the most common cause of dementia, AD afflicts about 7 million Americans as of 2024 and will affect 13 million Americans by 2050. The earliest neuropathology of AD is often the extracellular deposits of Amyloid-beta (Aβ) peptides. If not removed efficiently, Aβ causes synaptic dysfunction and starts to aggregate into plaques, which in turn deform neuronal processes, activate glial cells, and induce inflammation. Recent research has established Aquaporin 4 (AQP4), an astrocyte-specific water channel protein, as a key regulator of Aβ. The mechanism is debated: some researchers argue AQP4 facilitates Aβ removal, while others suggest it helps sequester plaques to minimize neuronal harm. Nonetheless, there is widespread agreement on AQP4's critical role in AD. Recently, we detected an isoform of AQP4, termed AQP4X, and showed that it facilitates the clearance of Aβ peptides and the remodeling of plaques. AQP4X arises from a rare phenomenon called ‘translational readthrough’, where about 20% of the translating ribosomes continue beyond the stop codon, generating an extended isoform of the protein. Unlike the normal-length isoforms of AQP4, which are located away from blood vessels, the extended AQP4X is found in specialized astrocytic projections called ‘endfeet’ that surround blood vessels. To investigate AQP4X in AD, we have developed a loss-of-function mouse (Aqp4No_X, extra stops added to abolish readthrough) and a gain-of-function mouse (Aqp4All_X, stop mutated to sense for constitutive readthrough). When crossed to an AD model (APP/PS1), Aqp4No_X shows impaired Aβ clearance and Aqp4All_X shows enhanced clearance, indicating the importance of AQP4X in Aβ removal. AQP4X may function through the glymphatic system, a pathway thought to involve perivascular AQP4 and the cerebrospinal fluid flow in the brain and remove Aβ from the interstitial fluid during sleep. Unfortunately, in AD, the perivascular pool of AQP4 is altered, and the glymphatic system is compromised. In this proposal, we test the hypothesis that AQP4X is a critical modulator of astrocytes’ response in AD pathology. Aim 1 focuses on identifying pathways relevant to AQP4X function, including the glymphatic pathway, meningeal lymphatics, and circadian rhythms, using techniques such as cerebrospinal fluid tracing and MRI. Aim 2 characterizes the effect of AQP4X on Aβ plaque morphology by quantifying changes in Aβ fibrillation, dystrophic neurites, astrocyte hypertrophy, and microglia activation. Finally, Aim 3 seeks to characterize the effect of Aβ plaques on AQP4X and astrocyte endfeet through biochemical, imaging, and endfoot-specific translational profiling. Overall, our proposal will illuminate the role of Aqp4 readthrough in AD pathology and may establish the phenomenon as a potential therapeutic approach to enhance perivascular AQP4 and to mitigate AD pathology.

NINDS - National Institute of Neurological Disorders and Stroke

ABSTRACT. Immunotherapy has emerged as a successful therapeutic strategy for a variety of cancers. The recent success of immunotherapies in other solid tumors has sparked increased attention to treatments targeting the immune system in glioblastoma (GBM) and other brain cancers. One of the key challenges in the successful treatment of brain tumors with immunotherapy is our lack of appropriate methods to visualize and quantify the killing of cancer cells by the immune system within the brain. Currently, contrast-enhanced magnetic resonance imaging (MRI) is used to evaluate treatment response and progression in these patients. However, the accurate determination of tumor progression from treatment-associated inflammation, remains an unmet clinical challenge. The lack of a useful response assessment has complicated patient care and the clinical development of these therapies. This proposal aims to address this by developing a novel imaging strategy to visualize and quantify the specific protein, known as perforin, which immune cells utilize to gain access to kill cancer cells. Herein, we will develop a first-in-class, small molecule positron emission tomography (PET) probe which is capable of passively crossing the blood brain barrier and binding to perforin, permitting differentiation between response to immunotherapy and non-response. This strategy would permit non-invasive visualization of perforin levels with minimal off-target activity as perforin is expressed exclusively by cytotoxic cells of the immune system. We will develop a library of novel fluorine-containing small molecules targeting perforin and advance the top 10 binding molecules for radiolabeling with fluorine-18 (SA1). We will then characterize their brain penetration, biodistribution, and stability and use quantitative benchmarks to advance the top 3 performing radiotracers for treatment monitoring studies (SA2). Lastly, we will assess the utility of perforin-PET to detect therapeutic response and predict outcomes in established syngeneic orthotopic mouse models of glioblastoma following treatment with immune checkpoint blockade (SA3). Success of this approach would allow for rapid translation and incorporation into clinical studies. This would permit clinicians and researchers to visualize and have real-time information of the killing of brain cancer cells by the immune system and make informed decisions regarding the effectiveness of immunotherapy for any particular patient. The significance of the proposed research is that it demonstrates a generalizable mechanism to monitor multiple types of immunotherapy in glioblastoma and other brain tumors (including pediatric brain tumors and brain metastases). Such work has the potential to improve response determination in brain tumor immunotherapy, spare unnecessary treatment side effects, and through this eventually improve the management of this disease.