Grants

Musica Sacra

a New Birth of Freedom

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.

NINDS - National Institute of Neurological Disorders and Stroke

Our long-standing goal is to understand how the brain resolves competing needs to determine optimal survival behavior. In this project we ask the question, can circuits activated in different need states suppress the activity of neurons that signal chronic pain? Thus, the experiments outlined will monitor and manipulate neural circuits that reduce chronic pain during competing need states. We focus on the lateral parabrachial nucleus (lPBN), which integrates ascending nociceptive information from the spinal cord with competing survival information. The activity of lPBN neurons is amplified in multiple injury models and activity of PBN neurons correlates with behavioral measures or responses to inflammatory or neuropathic pain. Our ongoing studies indicate that the lPBN receives a dense innervation from neuropeptide Y (NPY) neurons in the arcuate nucleus (that contributes to hunger, ARC), the subparafascilular nucleus (that contributes to fear, SFP), and the ventral periaqueductal grey (that contribute to thirst, vPAG). We show that hunger, thirst, and fear reduce stimulus-evoked activity of lPBNY1 neurons. Importantly, we find that blocking signaling at Npy1r (Y1) receptors in lPBN prevents the ability of hunger, thirst, and fear to reduce behavioral signs of neuropathic pain. This provides the premise for an overarching conceptual framework that Y1-expressing lPBN neurons (lPBNY1) serve as a hub for the integration of multiple needs states with chronic pain. Our research will define the neural inputs that underlie how internal needs (hunger and thirst) and external threats (fear) evoke the release of NPY (Aim 1A) that reverses the hyper-responsiveness of lPBNY1 neurons to sensory stimulation in mouse models of early inflammatory pain, latent sensitization models of chronic inflammatory pain, and nerve injury models of neuropathic pain (Aim 1B). We will use Y1 receptor pharmacology and mouse genetics to definitively demonstrate the cellular target of NPY in the lPBN (Aim 2) and will determine the functional role of each NPY input from the ARC, SPF, and PAG in mediating pain-response behaviors and negative affect associated with chronic inflammatory and neuropathic pain (Aim 3). These will be the first studies to implement assays of NPY release, microendoscopy lPBNY1 recordings, and conditioned place aversion to explore the neural circuits that determine how multiple needs engage distinct, previously unstudied NPY-expressing neurons to suppress chronic pain. Taken together, our experiments will chart the endogenous neural network that modulates the transmission of pain signals through the brain. Ultimately, these studies will reveal important insights into how the brain ranks and responds to competing survival needs in a dynamic environment.

NIA - National Institute on Aging

SUMMARY The prevalence of Alzheimer’s disease (AD), the most common form of dementia, is expected to triple and reach 13 million in the United States by the year 2050. Despite the public health burden of dementia on an aging population, the etiology of AD is still not well-understood. The interplay between the human virome—the total collection of viruses in and on the human body—and host immunity is linked to complex diseases, including AD. Evidence suggests that viral infectious pathogens, such as certain herpesviruses, promote the development of AD. However, methodological limitations of the relevant epidemiologic studies include: 1) reverse causality due to cross-sectional or retrospective case-control study design or consideration of viral infections in elderly participants; 2) underdiagnoses of infections due to detection of clinically apparent infections only; 3) confounding due to unmeasured infections; 4) lack of consideration of joint or interacting effects of multiple infections; and 5) lack of consideration of potential intermediate phenotypes, such as changes in brain and cognitive health, AD-related biomarkers, and DNA methylation patterns related to accelerated aging. The limitations of these studies render interpretation of their findings difficult, presenting an important research gap we propose to address. Recently developed high-throughput methods enable detection of immune responses to all human viruses. However, no studies conducted comprehensive analysis of antiviral antibodies in human sera for AD-related outcomes. We propose a prospective study that uses three prospective cohorts to identify viral infections associated with brain health, cognition declines, and the risk of AD later in life. We will use VirScan, a revolutionary new technology that uses bacteriophage immunoprecipitation sequencing (PhIP-Seq) for comprehensive serologic profiling of exposure history to all known human viruses. Our pilot work shows viral signatures are stable over time, supporting one-time measurement for long-term analysis. Our prospective case- control study of AD nested in the NYU Women’s Health Study (NYUWHS), a prospective cohort of 14,273 women (ages 35–65) who were enrolled between 1985–1991 and donated blood samples at baseline, will assess both: 1) responses to viral infections in relation to AD risk, and 2) whether viral infections are associated with epigenetic age acceleration and AD-related biomarkers. We will use the Cognitive Reserve (CR) study and the Reference Ability Neural Network (RANN) study, which share 529 healthy participants recruited since 2011 across the adult lifespan, to assess responses to viral infections in relation to longitudinal averages and changes in AD biomarkers, as well as brain and cognitive health measured using magnetic resonance imaging and neuropsychological tests. Our findings promise to discover emerging risk factors for AD, improve risk stratification, direct earlier intervention, and open new areas for prevention.

NYC Department of Cultural Affairs

A Public Space Literary Projects, Inc.

NIAID - National Institute of Allergy and Infectious Diseases

SUMMARY Infections caused by multidrug-resistant (MDR) Pseudomonas aeruginosa pose an urgent public health threat. MDR P. aeruginosa is a critical priority global pathogen for which new therapeutic approaches are desperately needed. At UPMC, over 100 patients each year develop serious infections with MDR P. aeruginosa; these infections are difficult to manage, frequently evolve resistance to last-line antibiotics, and are often deadly. There is an urgent need to develop new antimicrobial approaches for the treatment of MDR P. aeruginosa infections. Bacteriophage (phage) therapy is one such approach that has gained significant attention in recent years. We currently treat dozens of patients each year at both UPMC and across the country with phage therapy under compassionate use, which uses personalized phage cocktails to treat resistant bacterial infections in individual patients who have no other viable treatment options. Most of the patients we treat are suffering from MDR P. aeruginosa infections, and in the majority of patients treated thus far, phage therapy improved their clinical status. Despite this success, single patient anecdotes are not sufficient to rigorously test and develop phage therapy for widespread use. Here we propose to systematically assemble and evaluate a multi-phage cocktail targeting diverse MDR P. aeruginosa clinical isolates which can be paired with new β-lactam/β-lactamase inhibitor (BL/BLI) combinations currently used to treat MDR P. aeruginosa infections to improve antibacterial efficacy. Our central hypothesis is that a rationally selected cocktail of P. aeruginosa phages targeting contemporary MDR P. aeruginosa isolates will improve the activity of antibiotics commonly used to treat MDR P. aeruginosa infections. To test this hypothesis, we will leverage a large collection of contemporary MDR P. aeruginosa clinical isolates collected from 28 medical centers across the country as well as a panel of 10 lytic phages that have been previously approved for use in phage therapy to treat patients, and which were safe and well tolerated. In Aim 1, we will assemble a P. aeruginosa-targeting multi-phage cocktail that exhibits broad lytic activity against a national panel of 100 diverse MDR P. aeruginosa clinical isolates. In Aim 2, we will determine whether the combination of this P. aeruginosa-targeting phage cocktail and P. aeruginosa-targeting antibiotics results in superior and more sustained in vitro killing than either agent alone. Successful completion of these aims will establish a phage-antibiotic combination therapy that can advance in preclinical development and be used in an eventual clinical trial. The project will also directly inform the treatment of critically ill patients who cannot wait for a trial but could still benefit from receiving phage therapy under compassionate use.

Toledo Public Library

The library will upgrade the community meeting room by adding technology and hardware to encourage new uses such as small business conferencing space and community education opportunities.

NIDDK - National Institute of Diabetes and Digestive and Kidney Diseases

Project Summary Level 2 hypoglycemia (i.e., low blood glucose) can affect one’s daily life; generate fear and psychological distress; and lead to brain dysfunction, heart complications, and even death. People living with type 1 diabetes (T1D) are particularly susceptible to developing Level 2 hypoglycemia due to the obligatory use of exogenous insulin to manage diabetes. Advanced diabetes technologies (ADTs), such as continuous glucose monitoring and closed- loop insulin pumps, have become the standard of care to improve diabetes control and reduce hypoglycemia. Yet roughly one in three adults living with diabetes continue to develop Level 2 hypoglycemia despite using ADTs, and no strategies have demonstrated effectiveness in alleviating hypoglycemia beyond these standard-of-care technologies. Training to improve hypoglycemia symptom detection and psychoeducation for addressing unhelpful hypoglycemia beliefs have shown promise in reducing Level 2 hypoglycemia. However, these approaches’ effectiveness requires confirmation in ADT users. A scalable intervention with low patient burden is especially needed to expand access to these strategies. We have developed and feasibility-tested two automated real-time patient glucose data–guided digital intervention components delivered via SMS messages: digital hypoglycemia symptom detection training and digital hypoglycemia psychoeducation. In the proposed project, we will conduct a clinical trial to determine the individual and combined contributions of these two components to reducing Level 2 hypoglycemia in T1D adults who continue to develop Level 2 hypoglycemia despite using ADTs. We will then select the intervention that best balances effectiveness with recipient burden as the optimized program for future implementation. We will recruit 208 participants across the U.S. to achieve this objective. Under a 2×2 factorial design, participants will be randomly assigned to one of four experimental conditions: (1) digital hypoglycemia symptom detection training; (2) digital hypoglycemia psychoeducation; (3) both; or (4) usual care only; all participants will be followed up for 1 year. We will determine the individual and combined effects of the digital hypoglycemia symptom detection training and hypoglycemia psychoeducation on reducing Level 2 hypoglycemia (Aim 1). Then, we will identify and select the intervention that best balances effectiveness with recipient burden as the final program (Aim 2). Finally, we will investigate the intervention mechanisms of action and explore potential factors moderating the intervention effectiveness (Aim 3) based on our trial and qualitative interview data. We hypothesize that each digital component (i.e., hypoglycemia symptom detection training and hypoglycemia psychoeducation) will have an effect in reducing Level 2 hypoglycemia in adult ADT users. By the end of this study, we will (1) inform the strategies necessary to address Level 2 hypoglycemia beyond the current standard of care and (2) provide an effective, scalable solution to alleviate this devastating and even life-threatening complication.

U.S. National Science Foundation

A Science of Science Approach to Analyzing and Innovating the Biomedical Research Enterprise

Josephine County

A Street Building Petroleum Cleanup Project

NIAID - National Institute of Allergy and Infectious Diseases

Post-TB lung disease (PTLD) contributes substantially to the overall morbidity and mortality associated with TB infection. On a histopathological level, fibrosis is a significant feature of PTLD and likely underpins many associated symptoms and clinical findings. Adjunctive therapies that could be used with traditional anti-TB antibiotics to modulate tissue destruction and pathologic remodeling could improve post-treatment lung health, quality of life, and longevity for TB survivors. To date, our insight into the pathological processes driving TB- associated fibrosis is limited, consequently limiting the development of such adjunctive treatments. In the proposed work, we will begin to address that knowledge gap by creating a baseline understanding of the molecules and cells associated with fibrogenesis during TB infection. Our work will make use of unique resources in the Steyn group, including a Human Tissue Biobank of lung specimens that represent a range of TB- associated fibrotic pathologies, and the unique resources of the Barczak laboratory, who are applying a mouse model to study pathogenic mechanisms of TB-associated fibrosis. In Aim 1, we will use our mouse model and a combination of histopathology, immunohistochemistry, and immunofluorescence to identify spatial correlates of fibrogenesis. We will first test our hypothesis that defined macrophage subsets are spatially associated with fibrogenesis (1A). The idiopathic pulmonary fibrosis (IPF) mouse model is the canonical model for fibrogenesis in lung; we will next test whether molecular and cellular factors identified in the IPF model are associated with TB-associated fibrogenesis (1B). Complementing our hypothesis-driven work, we will use spatial transcriptomics to identify novel candidate molecules, cells, and pathways (1C). In Aim 2, we will use the Steyn lab Human Tissue Bank to benchmark findings from the mouse model and identify clinical drivers of TB-associated fibrosis. We will use metadata to identify clinical correlates of four intermediate-stage fibrosis morphologies (2A). We will then test associations between fibrosis and macrophage subsets (2B), additional molecular and cellular factors relevant in the IPF model (2C), and novel cellular and molecular factors identified in Aim 1C (2D). We will explicitly compare results between mouse and human specimens. We will then test associations between clinical factors and cellular and pathway correlates of fibrosis (2E). In Aim 3, we will use µCT and artificial intelligence approaches to develop algorithms that characterize and comprehensively quantify fibrosis in human lung sections and whole mouse lungs to identify candidate microanatomic contributors to fibrogenesis during TB infection. In addition to enabling a full comparison between mouse and human fibrosis, results of Aim 3 will enable the development of new hypotheses around microenvironmental and anatomical cues for fibrogenesis in TB infection. Successfully completing our three aims will create the foundational knowledge and novel tools necessary to ultimately build a mechanistic model for the path to TB-associated fibrosis and for preclinical testing of candidate interventions. We anticipate this work will directly contribute to new strategies for treating TB.

Forests, Parks & Recreation

A Trail for All at the Pratt Refuge

NIAID - National Institute of Allergy and Infectious Diseases

SUMMARY. Precise tissue- and temporally- specific manipulation of gene expression has driven mechanistic studies across many different model organisms. Bimodal expression systems such as the Q-system from N. crassa and the GAL4-system from S. cerevisiae are key genetic tools for elucidating gene function and cellular properties. These systems were introduced and optimized in Drosophila melanogaster, contributing to groundbreaking discoveries in embryonic development, animal growth, physiology, metabolism, and neuroscience, and have had a significant impact in the understanding of human biology, in both health and disease. Using CRISPR/Cas-9 technology, the Q-system (the QF2 driver and QUAS reporters) has been successfully introduced to other insects with major impact on human health, such as Aedes and Anopheles mosquitoes. Additionally, it has been applied to vertebrate model systems including the zebrafish Danio rerio. In contrast, while the GAL4 system, has been used in zebrafish, its transfer to mosquitoes has been unsuccessful due to cell toxicity. However, even the Q-system poses a number of limitations, including toxicity and the lack of an effective suppressor that functions independently of chemical compounds. In his application, the Q-system will be re-engineered to remedy these drawbacks by incorporating temperature-sensitive self-splicing intein modules (INTts), tested for efficacy in D. melanogaster and transferred to the zebrafish Danio rerio. Inteins are self-splicing endopeptidases embedded within proteins found in bacteria and unicellular eucaryotes (e.g., yeast), with the enigmatic property of excising themselves from pre-proteins, leading to the generation of a functional protein. Remarkably, inteins can be inserted into foreign proteins, where they self-splice as in their normal host protein. The experimental strategy leverages temperature-sensitive self-splicing inteins that have been validated in S. cerevisiae to self-splice at temperatures up to 270 C in foreign proteins. Specifically, inteins with permissive and restrictive setpoints between 15 and 300 C, compatible with growth and development of insects and fish, will be integrated into the DNA binding domain of QF2 (QF2_ INTts) and the protein interaction domain of QS (QS_ INTts), to disrupt their function when retained at restrictive temperature (24 to 300 C). When kept at permissive set points (17 to 230 C), intein removal via self-splicing activates QF and QS. Transgenic Drosophila will be tested for functionality of QF2_ INTts and QS_ INTts using QUAS-GFP reporter genes at different temperature. Genes encoding validated QF2_ INTts will be conferred to plasmids for D. rerio transgenic fish, tested for functionality using QUAS-GFP reporters. These new tools provide a much-needed temporal control element for the Q-system, making it invaluable for the research community to analyze gene and cell function relevant to human health.

First Hill Improvement Association

In Phase 2, the First Hill Improvement Association will continue working with Site Workshop (consultant), Parks, and DoN to lead the community through final design and construction documentation for improvements to First Hill Park. This phase of work will build off of the approved Phase 1 concept plan.

Friends of Small Business Saturday in Magnolia

The Magnolia Chamber of Commerce will engage the neighborhood to define a collective “Vision for Magnolia Village” and a set of actionable steps to help shape future development and growth.

Montgomery County MD / State of Maryland

Housing and Community Development

NIAID - National Institute of Allergy and Infectious Diseases

PROJECT SUMMARY Zika virus (ZIKV) is a re-emerging mosquito-borne flavivirus of significant public health concern. To date there are no effective licensed antiviral treatments or a vaccine. Therefore, elucidating the molecular biology of these viruses and the interactions with the host cell are foundational to identifying and developing effective treatment options. The single-stranded positive-sense RNA genome of ZIKV mimics a cellular mRNA. Specifically, the viral RNA encodes a single open reading frame, has structured untranslated regions (UTRs) adjacent to the open reading frame, and contains a N5’-methyl guanosine cap. Unlike mRNAs however, flaviviruses lack a poly(A) tail. From a multiple sequence alignment of all mosquito-borne flaviviruses we identified conserved regions containing 3-6 tandem adenosines in the 3’ UTR. These A-rich regions are localized in single-stranded regions within and between the XRN-1 resistant pseudoknots (xrRNA), dumbbell and 3’ stem loop (3’SL) RNA structures. In preliminary experiments we investigated the role of the A-rich region between xrRNA2 and the pseudo- dumbbell RNA structure (pre-pseudo/Y-dumbbell). Specifically, we generated three different mutations in a ZIKV Renilla luciferase reporter replicon and infectious clone. These mutants revealed that the pre-YDB A-rich region had a role in translation in both mammalian and mosquito cell lines. Different mass spectrometry studies have shown that the cellular poly(A) binding protein (PABP) interacts with the ZIKV 3’ UTR. Additionally, PABP was previously shown to interact with a region in the dengue virus 3’ UTR that harbors an A-rich motif. In preliminary studies we find that depletion of PABP1 decreased ZIKV, but not cellular, protein levels and viral titers without affecting cell viability. We therefore hypothesize that A-rich regions in the ZIKV 3’ UTR function to recruit cellular RNA binding proteins such as PABP1 to promote distinct steps in the virus infectious cycle. In Aim 1 we will mutate the A-rich regions in the infectious clone and a subgenomic luciferase reporter replicon to investigate the function of select A-rich regions on translation, replication, and viral fitness in mammalian and mosquito cells. In Aim 2, we will investigate the role of PABP1 on ZIKV gene expression and determine if PABP1 and other cellular RNA binding proteins bind A-rich regions in the ZIKV 3’ UTR. Overall, this study will advance our understanding of how flavivirus 3’ UTRs interact with the host to promote distinct steps in the infectious cycle in two vastly different hosts. Understanding how specific sequences in RNA genome function could lead to the therapeutic advancement namely the development of an attenuated vaccine ZIKV strain. Moreover, defining similar and unique RNA-protein interactions between mammalian and mosquito hosts could inform future vector control strategies.

Young Faces Smiling

Anti-Poverty

West Harlem Group Assistance, Inc.

Local

Montgomery County MD / State of Maryland

Housing and Community Development

HPZS

Management and Business Professionals and Administrative Services

Specialty Consulting, Inc.

Management and Business Professionals and Administrative Services

FGM Architects Inc.

Management and Business Professionals and Administrative Services