Search Grants — Free, No Account Required

NIA - National Institute on Aging

PROJECT SUMMARY Dementia, including Alzheimer’s disease (AD) and frontotemporal dementia (FTD), are major contributors to mortality, morbidity, and worldwide healthcare expenditure. FTD is fatal and incurable and represents 10-20% of all dementia cases. Approximately 9% of all FTD cases are caused by MAPT mutations (FTD-tau). Given that there are no effective treatments for FTD (an Alzheimer’s-related dementia), novel therapeutic strategies are urgently needed. Targeting the MAPT gene itself by CRISPR/Cas9 genome editing may provide a curative intervention. We have established a novel dual sgRNA strategy, which can excise the mutant MAPT allele in patient-derived induced pluripotent stem cells (iPSCs). The excision preserves expression from the non-diseased allele. In Aim 1, we will maximize efficiency of our gRNA strategy by identifying sgRNA pairs that excise the MAPT transcription and translation start sites on the mutant allele with high efficiency and no side effects in patient iPSCs and post- mitotic patient-derived neurons in vitro. We will then deliver our optimized editing reagents to an FTD mouse model (PS19) via AAV PhPeB, which cross the blood brain barrier and achieve brain-wide distribution. In Aim 2 we will optimize AAV dosing and determine whether CRISPR editing can reverse pathologic hallmarks of FTD- tau or only prevent their onset. In Aim 3 we will determine how three genes embedded in MAPT affect normal and pathologic tau expression, potentially providing new therapeutic targets, and in any case a useful context for any therapy that aims to alter tau expression or the MAPT locus. With the successful completion of these studies, we will have optimized a candidate gene editing strategy that targets the MAPT mutation, and reaches the highest therapeutic efficacy in human neurons in vitro. We will also determine the therapeutic window in vivo. Our editing strategy will then be ready to pair with human-specific delivery reagents that we and others are developing as they become available. We will have additionally addressed a number of open questions in the field, including whether editing efficiencies in post-mitotic neurons differ from mitotic cells, how to deliver CRISPR/Cas9 with multiple sgRNAs widely throughout the mouse brain, whether it is possible to reverse or arrest clinical phenotypes in symptomatic mice, and the impact of embedded genes on MAPT physiologic and pathologic function. This work will inform our understanding of normal MAPT function and provide proof-of-concept and IND-enabling studies for a novel MAPT CRISPR therapeutic. Our approach is likely also applicable to sporadic FTD-tau and other tauopathies, including progressive supranuclear palsy (PSP), Alzheimer’s disease (AD) and corticobasal degeneration (CBD). Our overarching goal is to accelerate genome editing for neurodegenerative diseases toward the clinic.

NIAID - National Institute of Allergy and Infectious Diseases

Cold Spring Harbor Laboratory Conference GENE EXPRESSION AND SIGNALING IN THE IMMUNE SYSTEM March 3-7,2026 ABSTRACT The proposed meeting on Gene Expression and Signaling in the Immune System, to be held in March, 2026, will focus on the most recent advances in this rapidly moving field. The meeting will be open, with attendance limited only by the facilities available to a maximum of ~425 participants. Oral presentations will be delivered by both invited speakers and those selected from submitted abstracts. This ensures the participation of junior and senior leaders in the field and the presentation of the most exciting results emerging at the time of the meeting. A particular emphasis will be made to ensure that a substantial number of speakers will be advanced trainees or junior PIs. The oral presentations will be complemented by poster presentations in two sessions, also selected from submitted abstracts. The areas to be covered in the 2026 meeting include 1) Regulation of gene expression; 2) Differentiation; 3) Signaling at the membrane; 4) Intracellular signaling; 5) Intercellular communication; 6) Host:microbe interactions; 7) Immune responses; and 8) Tissue-immune communication (immunophysiology). Rather than focusing on one particular type of immune cell or disease process, the meeting will highlight mechanistic approaches that aim to rigorously advance our knowledge of how the processes of signal transduction and gene regulation operate within the immune system at different scales. Ample opportunity is provided for the presentation of important, late-breaking findings. The meeting format ensures and encourages highly productive discussions, particularly during meals and in poster sessions. The meeting will foster interactions among immunologists working in related areas and provide a forum for the development of new ideas and approaches for current and future investigations of regulation of signaling and gene expression in the immune system.

Cornelius Public Library

Create an innovative and culturally responsive STEM (Science, Technology, Engineering, and Mathematics) Learning Lab to engage Cornelius youth in free and fun activities that promote the development of math and science skills needed for academic success.

Curry County Library Foundation

This Curry County Library Network project will upgrade Curry County libraries (Agness, Chetco, Curry Public, Langlois, and Port Orford) to a modern integrated library system, allowing them to provide current and updated library services such as faceted searching, online account management, automated notifications, and digital resource integration. Curry County libraries will enter into a cooperative agreement with Coos County and the libraries in both counties will share a single catalog and courier service. Together the two counties will be able to provide more comprehensive services, programming, and expanded online access to their rural populations.

Liquor World C/O Retail Serv & System (Total Wine)

Commercial

NIA - National Institute on Aging

Cytomegalovirus (CMV) is a common beta herpesvirus with over half of adults in the US infected by age 40 and higher seroprevalence among women than men. Though most acute CMV infection in immunocompetent individuals is mild or asymptomatic, following acute infection the virus remains in a latent state with the potential for reactivation across the lifecourse increasing risk of the creation of a chronic inflammatory state, a well appreciated component of accelerated biological aging. Chronic inflammation from any cause can lead to deleterious effects across all organ systems, resulting particularly in later midlife increased risk of metabolic and cardiovascular disease (CVD) in women. There has been mixed evidence implicating CMV seropositivity (CMV+) in the development of CVD and CVD-related mortality. A major limitation of many studies of CMV+ and CVD risk are that such studies do not adequately capture the critical window of physiologic and inflammatory changes that occur in women during the midlife. In the proposed K01, I will obtain the necessary training in aging and field methods to address these research gaps and pursue an independent research career. My research objectives are to a) describe the midlife prevalence of CMV+ within the SWAN cohort and relate CMV+ to subclinical cardiovascular outcomes; b) examine the effect of midlife inflammation on the relationship between CMV+ and carotid intima media thickness; and c) explore changes in IgG antibody level of multiple herpesviruses across the midlife, and to examine the relationship between individual-level change in IgG level and subclinical CVD outcomes. I hypothesize that that individuals with prior CMV infection will have worse subclinical CVD outcomes than those without CMV, particularly in the absence of other common inflammatory conditions such as morbid obesity and type 2 diabetes. To test these hypotheses, I will utilize longitudinal data and banked specimens from the Study of Women’s Health Across the Nation (SWAN), a multi-site cohort of midlife women transitioning into late adulthood. I will conduct this work at the University of Michigan School of Public Health, supported by an interdisciplinary research team that will guide my training in 1) aging and cardiovascular disease epidemiology; 2) immunology and immune-related aging; and 3) practical skills in the development and implementation of prospective data collection methods. By thoroughly evaluating the connections between CMV, midlife inflammation, and CVD we can improve preventative care by identifying those who might be at increased risk for CVD regardless of the presence of other risk factors. This research will also underscore the importance of nontraditional risk factors for cardiovascular disease in women. This grant is critical to meeting the National Institute on Aging’s strategic goal of better understand the biology of aging and its impact on the prevention, progression, and prognosis of disease, and the NIH Initiative on Women’s Health Research goal of developing personalized prevention strategies for cardiovascular disease in women. This award will help launch my independent research career evaluating the etiologic intersections of chronic and infectious disease.

DACRA TECH LLC

Information Technology Broadcasting and Telecommunications

North Country Library System

DEC Program in Jefferson, Lewis, and St. Lawrence Counties

NIAID - National Institute of Allergy and Infectious Diseases

Project Summary/Abstract The complexity of the HIV-1 transcriptome has been progressively revealed throughout recent decades using increasingly advanced RNA sequencing technologies. However, knowledge of the RNA primary sequence alone has not been sufficient to determine the importance or function of each of the over 40 highly conserved HIV-1 splice variants, which code for nine known proteins and polyproteins. RNA-protein interactions are fundamental to RNA fate and function. From transcription to cellular localization to translation of the gene product, and many steps in between, proteins interact with RNA to regulate gene expression and, in the case of HIV-1, viral replication. Notwithstanding the high significance of these splice variants in the HIV-1 life cycle, technologies for interrogating the functions, interactions, and cellular localizations of individual splice variants are woefully lacking. We propose to develop and validate a suite of powerful new tools to interrogate the functions, interactions, and cellular localizations of individual splice variants of HIV-1. For Aim 1, we will develop sensitive and multiplexed assays (HyPR-MS) to elucidate the protein interactomes of up to 20 conserved HIV-1 mRNA splice variants and advance mechanistic studies of newly identified viral RNA regulatory co-factors. In Aim 2, we will determine which HIV-1 splice variants are most critical to HIV-1 replication using virological assays and measure single- cell viral RNA abundance and subcellular localization using a multiplexed branched DNA fluorescence in situ hybridization technique (SV-FISH). These powerful new tools and strategies will be used to elucidate previously unobtainable information about HIV- 1 replication. Once developed, these same novel technologies will comprise a powerful new toolset that can be applied to splice variant investigations in other viral and cellular systems.

NIAID - National Institute of Allergy and Infectious Diseases

PROJECT SUMMARY This project aims to investigate the interactions and impacts between a depleting antibody (D-aPD-1) that targets programmed death-1-expressing (PD-1+) cells and the autoimmune environment in type 1 diabetes (T1D). PD- 1+ cells are primarily effector lymphocytes that are diabetogenic in T1D.These PD-1+ cells could be a potential target for suppressing autoimmunity in T1D. Initial studies demonstrated that eliminating PD-1+ cells with a PD- 1-specific immunotoxin delayed hyperglycemia onset in NOD mice with late insulitis (L-insulitis). However, immunotoxins are not ideal for chronic disease management, leading to the development of D-aPD-1 as an alternative. Interestingly, when administered to mice with early insulitis (E-insulitis), D-aPD-1 delayed hyperglycemia onset, suggesting a protective effect. However, when given to L-insulitis mice, it unexpectedly accelerated hyperglycemia onset, indicating a pro-autoimmune response. Furthermore, unlike D-aPD-1, PD-1 immunotoxin delayed hyperglycemia onset in L-insulitis mice. These findings align with clinical observations that disease stages and therapeutic agens influences treatment outcomes, highlighting the need to elucidate the mechanisms driving these contrasting responses for better desing and application of T1D therapeutics. To address these questions, this project will investigate the hypothesis that both the immune milieu of T1D and the characteristics of PD-1+ cell-depleting agents play key roles in determining treatment outcomes. Aim 1 will uncover the immune determinants of D-aPD-1’s divergent treatment outcomes in E and L-insulitis by profiling pancreatic islets and systemic immune cells before and after D-aPD-1 treatment. We will use single-cell RNA sequencing (scRNA-seq) and flow cytometry to dentify changes in immune cell populations and pathways associated with pro- and -anti-autoimmune effects. Additionally, strategies to shift L-insulitis responses toward an E-insulitis-like profile will be explored through modulate certain immune cell types. Aim 2 will define the mechanism underlying differential responses to D-aPD-1 and PD-1 immunotoxin by comparing immune cell dynamics and gene expression changes following the treatments. We will identify key cellular mediators responsible for the pro-autoimmune response induced by D-aPD-1, while also investigating whether depleting these responsive cell populations can mitigate its adverse effects. Aim 3 will establish that Fc-free PD-1+ cell depleting agents protect L-insulitis mice from hyperglycemia. Specifically, we will assess whether the Fc-free PD-1+ depleting agents, including albumin-amended antibody-drug conjugate (A3DC) and bispecific killer cell engager (BiKE), can eliminate PD-1+ cells without triggering pro-autoimmune effects. This research could advance targeted PD-1+ cell depletion strategies, leading to safer and more effective antibody-based treatments for T1D. By addressing immune context and therapeutic design, the findings may tailor the design and application of therapeutics for T1D patients at different stages.

NIDCR - National Institute of Dental and Craniofacial Research

7. PROJECT SUMMARY/ABSTRACT Although recent advancements in single cell and spatial transcriptomics have provided unprecedented resolution to study intercellular communication, there are currently still no comprehensive methods to fully decipher the complexity of truly functional cell-cell communication and to investigate its implications for human health and disease. The long-term goal is to reveal the complex networks of cell-cell communication to better understand their roles in maintaining tissue homeostasis, contributing to disease mechanisms, and ultimately guiding the development of novel therapeutic strategies. The overall objectives in this application are to 1) develop computational methods to systematically decipher functional cell-cell communication, and 2) investigate the regulatory roles of cell-cell communication in cellular states, tissue organization, and disease genetics. The rationale for this project is that while cell-cell communication is recognized as a fundamental aspect of tissue biology, its full complexity, especially in the context of disease, remains incompletely understood. By advancing computational methodologies and integrating multi-dimensional data, we can provide deeper insights into the mechanisms governing intercellular communication, which are crucial for both normal tissue function and disease progression. The central challenges will be addressed by pursuing three specific aims: 1) Develop innovative computational approaches to accurately define and categorize active cell- cell communication networks within tissues; 2) Systematically explore the impact of these communication networks on cellular states and tissue organization; and 3) Elucidate the functional contributions of cell-cell communication in the context of disease genetics. The research proposed in this application is innovative, in the applicant’s opinion, because it leverages cutting-edge computational and experimental techniques to integrate spatial and single-cell data, providing a multi-layered understanding of cell-cell communication. This approach allows for the creation of detailed communication maps and the identification of key regulators that could serve as targets for novel therapeutic interventions. This research is significant because it addresses a critical gap in our understanding of how cell-cell communication influences tissue function and disease systematically. By providing a comprehensive framework to study these communications, this project has the potential to transform our approach to understanding complex genetic diseases and to identify new therapeutic targets. Ultimately, such knowledge has the potential to offer new opportunities for the development of innovative therapies to treat genetic diseases, contributing to the broader field of precision medicine.

NIAMS - National Institute of Arthritis and Musculoskeletal and Skin Diseases

PROJECT SUMMARY Proper skin restoration after the damage is vital for organismal survival. The lymphatic vascular system is spread throughout the human body and has a critical function in mammalian physiology. In physiological conditions, its main function is the regulation of tissue drainage, immunosurveillance, and regeneration. Lymphatic dysfunction causes lymphedema a medical condition that manifests as tissue swelling and fibrosis. This serious condition results in profound severe delays in wound repair and the formation of chronic non-healing wounds. The mechanism by which lymphatic vessels regulate skin repair is unexplored. Additionally, while the role of lymphatic vessels in immune cell egress from tissues is well-established, how lymphatic vessels may directly modulate immune function in damaged tissues is poorly-defined. Recently, I discovered that lymphatic vessels are actively remodeled during wound healing to form small capillaries which are present in close localization to the wound front and hair follicles. This remodeling is critical for optimal repair as skin-specific loss of lymphatic vessels results in a significant delay in wound closure accompanied by a massive infiltration of immune cells. This proposal aims to leverage these observations by 1) delineating the mechanisms and consequences of lymphatic vessel remodeling during skin repair, and 2) determining the role of lymphatic vessels and fluid pressure in macrophage behavior during skin repair. This research stands to have a significant clinical impact because it can serve as a basis for developing new therapeutic avenues for lymphedema ulcers and chronic wound management. In addition, career-oriented guidance from my mentor and advisors, along with career development activated during the K99 phase that includes formal coursework on grant writing and project management, will further facilitate my transition to the R00 phase and my long-term productivity as an independent academic investigator. Collectively, the proposed research and career development plans are expected to generate data with a significant impact on understanding the repair and immunomodulatory functions of lymphatic vessels in skin repair and setting the basis of my future research as an independent researcher.

NIDDK - National Institute of Diabetes and Digestive and Kidney Diseases

ABSTRACT People with HIV-1 (PWH) on antiretroviral therapy (ART) are prone to experiencing chronic inflammation despite effective viral suppression. This sustained inflammation has been linked to an elevated risk of developing a variety of age-associated comorbidities, including chronic kidney disease (CKD). Preliminary data supporting this study demonstrates multiple distinct inflammatory endotypes among aging PWH on ART, several defined by levels of chemokine C-C motif ligand 2 (CCL2), a critical mediator and biomarker of kidney injury and disease. However, the precise cellular and molecular inflammatory immune endotypes in PWH that could lead to disease remain undefined. Moreover, how endotypes defined by circulating inflammatory markers impact organ- compartmentalized inflammation and the functional and molecular states of immune cells are unknown. The overall objective of this project is to define the early cellular and molecular signatures of inflammation associated with the progressive development of CKD in PWH on ART in both blood and urine. This will be achieved by comprehensively defining plasma inflammatory endotypes in a retrospective cohort of aging (50+ years) PWH on ART, sampled as they progressed from early to later stage kidney disease, with comparison to PWH on ART with normal renal function. In addition, systems immunology will be used to characterize the soluble and cellular inflammatory profiles of peripheral blood and urine in a prospective cohort of 200 aging PWH on ART. Urine, a readily accessible non-invasive biofluid, contains proteins and viable cells originating from the kidney that can serve as indicators of renal inflammation and overall kidney function. A combination of advanced machine learning approaches, clinical tests, and human kidneys-on-chips models will be applied to define the relationship between systemic, urinary, and renal cell inflammation and dysfunction in PWH on ART that are associated with onset of CKD. This project will test the hypothesis that specific inflammatory immune endotypes can be identified in PWH on ART that promote the activation and dysregulation of immune and kidney cells, contributing to the development of CKD. The hypothesis will be tested, and the overall objective achieved, with completion of three Specific Aims. Aim 1 will define plasma inflammatory endotypes of aging PWH on ART and identify signatures that predict CKD. Aim 2 will identify how plasma inflammatory endotypes impact the cellular, metabolic, and functional programs in blood and urine of aging PWH on ART. Finally, Aim 3 will determine the mechanisms of activation of renal inflammatory programs using kidneys-on-chips. This research will identify specific endotypes of inflammation associated with development pf CKD and uncover pathways driving chronic inflammation in the blood and urine that promote kidney injury and disease. This knowledge will enable the development of CKD risk prediction tools and of therapeutic interventions like CCL2 signaling inhibitors to reduce inflammation-related kidney disease and other comorbidities in this expanding population.

NIAID - National Institute of Allergy and Infectious Diseases

PROJECT SUMMARY/ABSTRACT Systemic lupus erythematous (SLE) causes substantial mortality and morbidity. Current treatments are not highly effective in modifying disease progression, especially in severely affected patients with renal involvement. Hence, there is a large unmet need to develop novel therapeutic targets. Genome-wide association studies (GWAS) offer an unbiased approach to identify genes that play an important role in driving disease pathogenesis. To identify risk genes in immune cell types relevant to SLE pathogenesis, we performed the first large-scale single-cell eQTL study on activated CD4+ T cells and found that SLE-risk variants were strongly associated with increased expression of the poorly understood gene ILRUN. Our study provides the first direct evidence that activated CD4+ T cells are the key cell types in which SLE-risk variants increase the expression of ILRUN. In Aim 1, we will identify the functional SLE-risk variants associated with ILRUN expression in activated CD4+ T cells. We will employ CRISPRi assays to determine functional enhancers that overlap ILRUN eQTLs, perform luciferase reporter assays to determine functional variants in ILRUN promoter and enhancers, and perform ChIP assays to identify the functional ILRUN eQTLs that directly perturb the binding of key transcription factors and modulate ILRUN expression. In Aim 2, we will assess the functional role of ILRUN in CD4+ T cells in the context of SLE and determine whether ILRUN influences the activation, apoptosis, proliferation, differentiation and cytokine production of CD4+ T cells. Overall, the studies examining the expression, regulation and function of ILRUN in CD4+ T cells will provide important mechanistic insights into the genetic basis of SLE.

NINDS - National Institute of Neurological Disorders and Stroke

PROJECT SUMMARY Neural circuit development and function depends on precise interactions between neurons and glia. Astrocytes, the primary peri-synaptic glia, mediate synapse formation, stability, and function. Neuron-astrocyte crosstalk is facilitated by complex protein-protein interactions, and loss of these interactions contributes to circuit instability in many neurological disorders. Thus, understanding the mechanisms that regulate neuron-astrocyte communication is of broad clinical importance. Glycosylation is a posttranslational modification that regulates protein stability and binding through addition of sugar groups to specific amino acids. Mutation of genes in glycosylation pathways cause congenital disorders of glycosylation (CDGs), a group of monogenic disorders associated with neurological dysfunction, including epilepsy, autism, and cerebellar degeneration. The mechanisms underlying neurological dysfunction in CDGs remain unknown. Here, I focus on ALG8, an enzyme in the N-glycosylation pathway. To explore the molecular underpinnings of ALG8-CDG, I first needed to develop models that reflect the patient population. To this end, I generated a predicted null zebrafish line (alg8stl973) and human embryonic stem cell (hESC) lines with a missense mutation (p.Thr47Pro) found in ALG8-CDG patients. My preliminary data revealed a decrease in astrocyte numbers in the brains of alg8 mutant zebrafish with no change in total cells, and reduced proliferation of ALG8 mutant hESC-derived astrocytes. Moreover, in alg8stl973 fish, astrocyte morphological complexity is reduced. As astrocyte-synapse association is necessary for neuronal signaling, I hypothesize that defective glycosylation disrupts specification and maturation of astroglia, which in turn drives circuit imbalance and CDG-associated behavioral deficits. To address this hypothesis, I will leverage preexisting transgenic tools in zebrafish to label astrocytes and test whether changes in proliferation and/or cell death result in reduced astrocytes in alg8stl973 fish (Aim 1). Furthermore, I will use biochemistry and in vivo imaging to characterize how loss of alg8 impacts the glycosylation status of one key regulator of astrocyte morphogenesis: NrCam (Aim 2). Finally, as ALG8 is expressed in all neural cell types, I will use cell-type specific rescue in fish and co-culture of hESC-derived neural cells to determine which cell type(s) drive changes in astrocyte morphology and synaptogenesis in ALG8-CDG (Aim 3). My long-term goal is to define common molecular changes in brain development across distinct CDGs. Critically, various CDG subtypes result in common neurological symptoms, but the cellular and molecular underpinnings of these phenotypes are largely unknown. Similar to my preliminary findings in ALG8-CDG models, recent work indicates that astrogenesis is altered in a mouse model of MGAT5-CDG, a CDG with defective N-glycosylation. Thus, I anticipate that my findings will be broadly applicable to the CDG community and will enhance our fundamental understanding of how glycosylation shapes brain development.

NIAID - National Institute of Allergy and Infectious Diseases

Inflammatory Bowel Diseases (IBD), including Crohn's disease (CD) and ulcerative colitis (UC), are life-long diseases characterized by cycles of active flares and remission. Disease flares result in significant inflammation in the colon, as well as pathology in other organ systems, including the brain. Central nervous system (CNS) dysfunction is a well-established comorbidity in patients with IBD. We have recently demonstrated CNS inflammation, and specifically inflammasome activation, is induced in the CNS of mice in an acute colitis model using a novel mouse model in which inflammasome activation induces tissue bioluminescence. In this grant, we seek to understand the mechanisms by which CNS inflammation is induced in this and similar models. We seek to extend exciting preliminary data which reveal that colitis results in the presence of a factor or factors in serum that drive CNS inflammation to identify this factor and the mechanism by which is induces inflammasome activation in the CNS. We also propose to use a newer version of our mouse model, in which expression of our luciferase based caspase-1 biosensor is restricted to specific cell lineages in the CNS, allowing us to understand the impact of colitis and CNS inflammation on specific cell populations.

NIAID - National Institute of Allergy and Infectious Diseases

Project Summary Acari (mite and tick)-borne illnesses severely impact human health, and it is essential to understand the mechanisms maintaining the pathogens within their vectors for the development of countermeasures. Transovarial transmission (ToT) is used by mites and ticks to keep vector populations persistently colonized, but virtually nothing is known about the mechanisms of vertical transmission. While most work focuses on ixodid ticks, mites and argasids are understudied vectors most likely because of their complex biology. We developed the argasid–relapsing fever (RF) spirochete model (Ornithodoros turicata–Borrelia turicatae), which is one of the most comprehensive systems to study reproduction and ToT. We reported O. turicata vertically transmits B. turicatae by autogenous reproduction (laying of eggs without a blood meal), a unique aspect of argasid reproduction. This means that endemic foci of RF spirochete-infected ticks can be established quickly without needing a blood meal host. We have also identified an isolate of B. turicatae that is infectious by tick bite but fails to be vertically transmitted to offspring ticks. This phenotype enables us to study the mechanisms of ToT in this complex non-model system through comparative genomics. Also, our work in O. turicata genomics identified vitellogenins and the vitellogenin receptor, proteins that can be targeted by plant and veterinary microbial pathogens for ToT. Building on prior work and our developed entomological, bacterial, genomics, and genetics resources, we can now identify the molecular players involved in ToT. This application implements a functional genomics approach to test the hypothesis that ToT of RF Borrelia occurs throughout the reproductive cycles of female ticks and is driven by B. turicatae binding to vitellogenins and/or the vitellogenin receptor. The first aim will define the reproductive cycles of O. turicata and assess the dynamics of ToT for B. turicatae. This will be accomplished through the utilization of developed in vitro and in vivo tick feeding systems, tick colonies, and a diverse collection of B. turicatae isolates. The second aim will identify the molecular constituents of ToT. We identified gene loci and plasmids associated with a vertical transmission phenotype. We will build on these findings through comparative genomics and transcriptomics and identify B. turicatae surface proteins that bind to vitellogenins and/or the vitellogenin receptor. Using our developed genetics, we will transform the non- vertically transmitted B. turicatae isolate with plasmid and/or gene candidates to restore a ToT phenotype. While rickettsial, viral, parasitic, and RF Borrelia undergo ToT in mites and ticks, the molecular mechanisms are unknown. The completion of this project will result in the first identification of the molecular constituents involved with ToT of an Acari-borne pathogen. These findings will likely be broadly applicable and move the field closer to finding interventions that disrupt the life cycle of pathogen and vector.

NIDDK - National Institute of Diabetes and Digestive and Kidney Diseases

PROJECT SUMMARY The complex communication mechanisms involved in bladder function are not fully understood. Due to this lack of physiological understanding, bladder diseases have remained challenging to treat, resulting in severe detriment to patient quality of life, particularly in the case of interstitial cystitis/bladder pain syndrome (IC/BPS). To address this, I aim to contribute to the ongoing investigation of the role of renin-angiotensin signaling in the bladder to inform future therapeutic strategies. Angiotensin II (Ang II) is the primary effector molecule of the renin-angiotensin system, and action on either of its receptors, Ang II type 1 receptor (AT1R) and Ang II type 2 receptor (AT2R), has been shown to elicit major effects in other organ systems. While Ang II signaling research in the bladder has primarily focused on defining the role of AT1R in mediating contractions and pathological effects on bladder tissue, AT2R should also be considered for its potential protective effects. Despite the understanding that AT2R counteracts the pathological impact of AT1R in other organ systems, the role of AT2R in the bladder remains largely unknown. Preliminary investigations have demonstrated that AT2R is expressed in the bladder, and one investigation showed that blocking AT2R in mice worsened the pathological effects in a cystitis model. However, there has not been a comprehensive investigation solely regarding the presence and role of AT2R in the bladder. Given these preliminary findings, I hypothesize that AT2R has a functional role in the bladder, and its activation can alleviate hyperalgesia and bladder dysfunction associated with cystitis conditions. This work will be conducted with two main aims: Aim 1 will provide a physiological evaluation by using naïve mouse bladder tissue to characterize the presence and location of AT2R mRNA expression and define the role of AT2R in ex vivo contractile responses through pharmacological manipulation of AT2R activity; Aim 2 will explore the role of AT2R in a pathophysiological bladder condition in vivo through chronic administration of an AT2R agonist, C21, in a cystitis mouse model. The effect of AT2R activation will be evaluated through voiding function, abdominal hypersensitivity, molecular changes, and ex vivo contractile responses. This work begins to uncover the role of AT2R in bladder physiology and pathology and is the first to broach the potential of AT2R as a therapeutic target for bladder diseases such as IC/BPS. Expanding the knowledge of angiotensin signaling in bladder dysfunction provides a foundation for pharmacological and non-pharmacological interventions that could improve patient outcomes and quality of life. The proposed research will help me develop as an independent scientist, providing me the opportunity to lead an experiment and gain the technical skills needed for a career in science. As a PhD student in the Joint Department of Biomedical Engineering at the Medical College of Wisconsin and Marquette University under mentorship from Dr. Aaron Mickle and Dr. Justin Grobe, I have all the tools necessary to complete this fellowship effectively and obtain preparation for my career.

NIAID - National Institute of Allergy and Infectious Diseases

PROJECT SUMMARY Non-typhoidal Salmonella enterica (NTS) is a leading cause of bacterial foodborne gastroenteritis and leading cause of death amongst foodborne pathogens. NTS elicit neutrophilic inflammation both during gut colonization and in systemic infection. Neutrophils produce toxic products to kill invading pathogens. One toxic product produced by activated neutrophils during severe bacterial infection is sulfite. Sulfites are reactive sulfur species that are toxic to all life forms. In addition to their presence during severe bacterial infection, sulfites are endogenously produced during metabolism of sulfated amino acids and are ingested in food products. Sulfites are also added to food products as antimicrobial agents. Therefore, enteric pathogens such as NTS must resist sulfite toxicity in several stages of infection. Our preliminary data link yeiH with sulfite stress resistance in Salmonella Typhimurium. We show a ∆yeiH mutant is defective for growth in sulfite stress conditions and yeiH expression is specifically activated by sulfite. yeiH encodes a highly conserved putative inner membrane protein with no documented function. The purpose of the proposed work is to establish the role of YeiH in sulfite stress resistance. We hypothesize that YeiH exports sulfites to allow Salmonella to resist neutrophil-derived sulfite stress during infection. We will test our hypothesis in two aims. In Aim 1, we will establish the role of yeiH in infection. We will use animal models of colitis and sepsis to establish whether yeiH is needed for Salmonella fitness during infection. Using a yeiH transcriptional reporter as a novel sulfite biosensor, we will determine the tissues in which yeiH is expressed to establish the spatial organization of sulfite stress during enteric and systemic salmonellosis. In Aim 2, we will determine how YeiH contributes to sulfite stress resistance. We will determine whether the proton motive force drives YeiH function and establish the amino acid residues important for YeiH role in sulfite stress resistance. We will also establish whether YeiH function is to export sulfite from the cell. YeiH orthologs are present in many bacteria, including in different pathogens with importance to human health. Successful completion of the proposed experiments will allow us to establish how YeiH contributes to Salmonella sulfite stress resistance in vitro and in vivo. This work will lead to future studies to define the mechanism of YeiH function and establish how sulfites contribute to human health and resistance to bacterial infections.

NIAID - National Institute of Allergy and Infectious Diseases

Project Summary Pathogenic bacteria have developed an array of strategies to undermine the host's defense mechanisms. In the context of non-Typhoidal Salmonella Typhimurium, a notable strategy involves the confinement of a single bacterium within a host vacuole called the Salmonella Containing Vacuole (SCV). This approach potentially provides an evolutionary advantage: by ensuring one bacterium per SCV, the host must target each SCV individually to eliminate the bacterial load, as opposed to confronting a vacuole harboring multiple bacteria clustered together. This mechanism could potentially extend the time required to combat the infection. Moreover, a single bacterium residing within an SCV exploits all accessible nutrients for replication and division. Conversely, multiple bacteria clustered within a single vacuole may engage in nutrient competition. Hence, understanding the fundamental mechanism of bacterial division in conjunction with vacuolar scission is imperative for gaining deeper insights into the pathogenic strategies employed by Salmonella enterica. Prior to this proposal we engineered a series of genetically minimal pathogenic strains that eliminates redundancy within the complex SPI-2 effector gene repertoire of Salmonella enterica serovar Typhimurium. Using this unique resource, here we will determine how a small network of SPI-2 T3SS effector proteins coordinate the complex events involved in SCV membrane scission and bacterial division within the host cell. This includes determining the location and host substrates of effector proteins at the SCV membrane using single cell particle tracking and live cell imaging (Aim 1). We will also investigate the molecular mechanisms of individual effector proteins that target a novel Rab-family GTPase defense system of the host (Aim 2). The resulting cellular and biochemical theories will be tested in murine models of systemic disease that are designed to evaluate effector protein functions at single cell resolution (Aim 3). Developing new drugs that target bacterial effector – host enzyme complexes would be an innovative approach to combat emerging infectious disease. While this idea holds great potential, the paucity of mechanistic information gleaned from deep studies into virulence factor functions has so far hampered their development as suitable drug targets. As a means to this end, the work performed here will allow us to predict new mechanisms of action for understudied Salmonella effector proteins and provide a glimpse into the structural-based evolutionary progression of a related pathogen groups.

Treasury Department

Pursuant to the Payment Integrity Information Act of 2019 (PIIA) (31 U.S.C. 3351 et seq.), the U.S. Department of Treasury (Treasury) operates the Do Not Pay Working System--a centralized portal through which agencies can search multiple databases to verify payment or award eligibility. Treasury, under a delegation from the Director of the Office of Management and Budget (OMB), is authorized to designate new databases for inclusion in the Do Not Pay Working System. Treasury is issuing this Notice of Proposed Designation to provide the public an opportunity to comment on the proposed designation of the Social Security Administration's Numerical Identification System (Numident) to the Do Not Pay Working System.

NIAID - National Institute of Allergy and Infectious Diseases

SUMMARY Autoimmune type 1 diabetes (T1D) is caused by the T cell-mediated destruction of insulin producing beta (β) cells in the pancreas. The incidence of T1D is rising globally. This is thought to be linked to early life exposure to microbes, environmental pollutants, and even diet. Viral infections in mouse models of diabetes have shown both acceleration and protection of diabetes, but these differences occur at different ages and degree of insulitis. The deciding factor in whether CD4+ T cells will prime the immune system to initiate T1D is the inflammatory context of the initial peripheral antigen encounter, particularly the timing of type I interferon (IFN-I) exposure triggered by microbes. Unfortunately, the vast majority of research has utilized specific pathogen free (SPF) mouse models examining activation in the absence of IFN-I. These conditions are very different from the environment in which humans live. At UMN, we have created a ‘dirty’ mouse model or normal microbial environment (NME) to study how the immune system responds or develops in the presence of microbes and viral pathogens, a more physiological environment driven by IFN-I production. Thus, we now have a diabetes model that we can study with NME conditions. Depending on the age and duration of time in NME, our NOD mice can be completely protected from diabetes. Our goal is to understand how infections impact immunity to either trigger or protect against diabetes. A better understanding of this process could provide therapies that prevent or treat human diabetes. We will test three specific aims; 1) Determine the role of IFN-I on naïve CD4 T cell fate following TCR activation in SPF conditions, 2) Determine the mechanism(s) by which NME prevents autoimmune diabetes, and 3) Develop autoimmune diabetes therapies. Our goal is to determine the role of IFN-I during initial T cell fate decision between Teff and Tregs. We hypothesize this fate decision leads to iTregs and will focus on induction, survival or enhanced suppressive function. Finally, we will explore IFN-I therapy and engineered TCR Tregs for autoimmune therapy.

NIAID - National Institute of Allergy and Infectious Diseases

Project Summary Pneumonic plague is the deadliest form of disease caused by Yersinia pestis. Disease manifests as a rapidly progressing and highly lethal necrotic pneumonia that is typically fatal within seven days. If antibiotic treatment is not administered within 24 hours after the onset of symptoms, pneumonic plague is 100% fatal. The lethality of plague is due to the abrupt onset of a hyper-inflammatory response that compromises pulmonary function. Deletion of the Y. pestis inner membrane protein YbtX results in decreased neutrophil infiltration into the airways and decreased expression of select pro-inflammatory cytokines/chemokines during the later stages of pneumonic plague. YbtX is part of a zinc acquisition system by which the Y. pestis siderophore Yersiniabactin (Ybt) competes with host protein calprotectin to scavenges bioavailable zinc. YbtX acts as an inner membrane transporter that facilitates transport of Ybt-Zn into the bacterial cytosol. Calprotectin is also known to be a driver of host inflammatory responses. We hypothesize that YbtX-mediated zinc import contributes to the onset inflammation in the lung by activating calprotectin-dependent inflammatory responses. To test this, we will characterize how zinc depletion impacts inflammation during infection in the absence of YbtX. Further, we will alter the balance of host zinc/calprotectin in a murine intranasal infection model of pneumonic plague and characterize how this balance affects pulmonary inflammation. This work will determine how zinc and host calprotectin impact host inflammatory responses during pneumonic plague.

NIA - National Institute on Aging

Project Summary Anti-aging antibody research, including strategies targeting interleukins and other antigens, shows promise in rejuvenating the immune system, improving metabolic functions, and extending healthy lifespans. AI-driven platforms are revolutionizing antibody development by accelerating affinity maturation and optimizing developability properties, enabling simultaneous optimization of multiple characteristics. These advancements could lead to more effective treatments for age-related diseases and a significantly improved quality of life for the growing aging population. However, zero-shot predictions for antibody affinities using pretrained models without additional target-specific data remain challenging. In this project, we propose a new strategy to address this challenge by generating diverse antibody-antigen interactions at an unprecedented scale (Aim 1) and training new AI models using these generated data in combination with data collected from literature and public databases (Aim 2). We will rigorously evaluate the performance of the new models and benchmark against the state-of-the-art methods. We will test the generality of the new models on a diverse set of antigens and experimentally validate the prediction accuracy (Aim 3). We will apply the models to identify new antibodies against new therapeutic targets associated with ageing or age-related diseases. Once complete, the proposed research will provide a powerful tool for accelerating antibody discovery and optimization as well as new antibody candidates for anti-aging treament.