Search Grants — Free, No Account Required

NINDS - National Institute of Neurological Disorders and Stroke

Project Summary Brain motor circuitry is influenced by neurohormonal regulators that underlie energy homeostasis through regulation of food intake (satiety) and physical activity. Leptin is a key player in these incompletely understood interactions. We have shown that leptin boosts striatal dopamine release by increasing the excitability of striatal cholinergic interneurons (ChIs) that regulate dopamine release via activation of nicotinic acetylcholine (ACh) receptors on dopamine axons. We also find that leptin directly excites dopamine neurons of the substantia nigra pars compacta (SNc). Moreover, systemic leptin enhances locomotor behavior in open-field testing. Given the role of the nigrostriatal pathway in motor regulation, highlighted by the motor deficits of Parkinson’s disease, which is characterized by loss of SNc dopamine neurons, our findings suggest that nigrostriatal dopamine is the neural substrate for leptin’s motor enhancing effect. Key aspects of this regulatory process are missing, including how leptin activates ChIs and SNc dopamine neurons, and whether motor activation by leptin is mediated at the level of the striatum or SNc, or both. In Aim 1, we will focus on the cellular level by identifying ion channels involved in ChI and SNc DA neuron activity. Experiments will test possible closure of two different K+ channels (Kv1.3 and TASK-3) as the mechanism by which leptin excites these cells using current-clamp recording in ex vivo striatal and midbrain slices. We will also examine the influence of these channels on evoked dopamine release in slices, monitored using fast-scan cyclic voltammetry. In Aim 2, we will move to the circuit level and determine the extent of dopamine and ACh release in dorsolateral striatum in vivo after leptin administration, with dopamine and ACh detection using genetically encoded sensors (GRAB-DA and GRAB-ACh) with fiber photometry. A key experiment will be to determine whether the motor-enhancing effect of leptin is absent in mice lacking ACh synthesis in striatum. Overall, the cellular mechanisms and circuits examined in this project highlight the role of leptin as a novel regulator of motor output, which may have implications for alternative therapies in Parkinson’s disease, while leptin’s DA-enhancing effects may also be relevant for depression and other neuropsychiatric disorders that are linked to dopamine dysregulation.

NIAID - National Institute of Allergy and Infectious Diseases

Project Summary The overarching goal of this R21 proposal is to define the mechanisms by which Group A Streptococcus (GAS) senses and adapts to changing heme levels encountered during invasive infections such as bacteremia. Heme is both a critical nutrient for GAS that is kept sequestered by hemoproteins in the host as well as a toxic molecule when in excess such as following hemolysis by GAS in the blood. We hypothesize that GAS carefully monitors the levels of heme in both the extracellular and intracellular compartments and accordingly regulates the expression of heme efflux, detoxification and repair pathways. GAS likely uses heme-sensing regulatory proteins, such as two-component signal transduction systems (TCS), to coordinately control the expression of a heme defense program. In this application, we will combine molecular genetic and biochemical methods with transcriptome studies and genome-wide mutagenesis screens to identify and characterize GAS mechanisms for heme sensing and regulation as well as establish their role in GAS pathogenesis. Our preliminary RNA-seq analysis of heme stress revealed induction of sugar utilization operons for fructose, lactose and galactose that are conserved with the published responses to iron-mediated stress. Here, we expand on our published and preliminary studies to define the molecular mechanisms used by GAS to deal with heme as both a nutritional signal and a challenge during infection: Aim 1 will determine whether TCS sensor kinases are required for GAS adaptation to heme toxicity by RNA-seq and use Tn-seq to identify key regulators required for heme management. Aim 2 will investigate the regulation of a GAS sugar metabolic shift during heme stress and ask why GAS induces these metabolic pathways to protect against heme. Finally, Aim 3 will assess the role of heme-regulated signaling through TCS sensor kinases or other pathways using in vitro, ex vivo, and in vivo models of GAS bloodstream infection. This project will help reveal the fundamental impact of heme in the pathophysiology of a major human pathogen. Completion of the proposed aims will provide novel insights into the host survival strategies used by GAS and related pathogens, and will set the stage for the development of new interventions that target bacterial heme management mechanisms.

NIAID - National Institute of Allergy and Infectious Diseases

Project Summary/Abstract: Hypervirulent Klebsiella pneumoniae (hvKp) poses an urgent public health threat as a cause of severe infections, with growing concern due to emerging multidrug resistance. While intestinal colonization precedes most hvKp infections, the regulatory mechanisms controlling this crucial step remain poorly understood. Our preliminary data reveal that host-derived environmental cues in the gastrointestinal tract regulate hvKp capsule production and virulence genes, potentially orchestrating the transition from colonization to systemic infection. This R21 proposal will investigate how host signals coordinate hvKp gene expression during intestinal colonization and dissemination. We will define how these environmental cues regulate capsule production and virulence gene transcription through RNA sequencing and genetic approaches, focusing on their effects on virulence-associated phenotypes and host-pathogen interactions. This developmental research addresses a fundamental gap in understanding hvKp pathogenesis. By uncovering host-responsive pathways that orchestrate colonization and virulence, we will reveal novel therapeutic strategies against multidrug- resistant and hypervirulent K. pneumoniae, supporting future mechanistic studies and opening new avenues for preventing and treating life-threatening infections.

Asian Americans for Equality, Inc.

Installation of new windows, insulation, boiler and hot water heating systems, roof work, exterior pointing and waterproofing, and various apartment rehab. Low flow shower heads, Energy Star appliances and other upgrades to enhance efficiency of utility

Rural Business-Cooperative Service

Renewable Energy Systems and Energy Efficiency Improvements Program

Waterbury Youth Services System, Inc

Renovation

Bridges...A Community Support System, Inc.

Renovations to building

Bridges...A Community Support System, Inc.

Renovations/Open Door Social Club

Chester

Replaced Boilers at Town Hall and complete new HVAC system at Public Works Garage

Seattle Polish Foundation

This project is a replacement of the HVAC system at the Polish Cultural Center Dom Polski located at 1714 18th Avenue. The existing HVAC system has stopped working properly causing the rooms to remain cold in winter and without adequate ventilation during the summer. The replacement of the HVAC system will allow the community to continue to use this special space for Polish cultural events and events for the local community like the National Night Out, Neighborhood Block Parties, and other programs. This project will be completed by March 31, 2023.

North Haven

Replacement of Library Chill System

Dept of the Army -- Materiel Command

REQUEST FOR INFORMATION – MUTUALLY BENEFICIAL MULTIDISCIPLINARY INNOVATION ECOSYSTEMS

Naval Supply Systems Command

Research Initiatives at the Naval Postgraduate School

Oakridge Public Library

Purchase and set-up equipment to host a Documentary Film Festival for the community.

La Mama Experimental Theatre Club, Inc.

Complete Interior Renovation of landmarked façade will be restored and the sidewalk and underground vault will be rebuilt. A mechanical bulkhead will be added. Two theatre spaces will be renovated with ne theater systems for rigging, audio, video, improve

Asian Counseling and Referral Service

Provide a Retail Technology Class to persons with mental illness. Twenty students will learn iPad Point-of-Sale (POS) systems, computerized cash registers, inventory management, and sales reporting. Students will practice their newly acquired retail skills.

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

RETAINER, STEM

Finance, Office of

RFP - Request For Proposal

Rhode Island Infrastructure Bank

Clean Water financing supports municipalities and utilities in upgrading water and wastewater infrastructure, improving water quality, and ensuring safe drinking water systems across Rhode Island communities.

ORIGAMI RISK LLC

Information Technology Broadcasting and Telecommunications

NIAID - National Institute of Allergy and Infectious Diseases

PROJECT SUMMARY Tumor necrosis factor (TNF)-α is a major inflammatory cytokine involved in the pathogenesis of several autoimmune and inflammatory diseases, including rheumatoid arthritis (RA). Current approaches aim to curb TNF-α-induced inflammation and tissue damage by treating patients with TNF-α inhibitors (specific antibodies or soluble receptors). The ineffectiveness of TNF inhibitors in >40% of patients partly indicates that we do not yet fully understand the underlying signaling mechanisms to effectively target TNF-α. While the use of TNF inhibitors has provided new insights into human immune and inflammatory systems and the mechanisms involved in disease processes, adverse events, and the re-emergence of the disease upon cessation of therapy suggest that other pathways might be involved in re-establishing the disease. Previous studies from our lab using RA synovial fibroblasts (RASFs) and preclinical models of RA not only shed light on the mechanisms by which TNF- α utilizes cell surface or cellular proteins to cause progressive inflammation and tissue destruction but also provided novel pharmacological approaches to suppress TNF-α’s function in RA. In this proposal, our novel preliminary data show that TNF-α utilizes Fn-14 (fibroblast growth factor-inducible 14), a receptor originally characterized for mediating TWEAK cytokine signaling. Knockdown of Fn-14 significantly reduced TNF-α- induced RANTES, MCP-1 and MMP-1 production, and cellular expression of podoplanin and cadherin-11 in human RASFs. In Fn-14-overexpressing cells, even low TNF-α concentrations synergistically induced inflammation, suggesting a potential undescribed mechanism exploited by TNF-α to propagate inflammation. RNA-sequencing analysis revealed >200 differentially expressed genes (DEG) affected by Fn-14 knockdown in TNF-α stimulated RASFs. Gene set enrichment analysis (GSEA) on the RNA-seq data revealed that IFN-α and IFN-γ pathway responses were significantly altered. Intraperitoneal administration of Fn14 antagonist (L524- 0366; 10 mg/kg; daily from the disease onset) inhibited collagen antibody-induced arthritis in DBA1/J mice. Based on these novel observations, we hypothesize that TNF-α utilizes the Fn-14 receptor as a non-canonical signaling pathway to induce inflammation. Therefore, in specific aim 1, we plan to determine the molecular mechanisms through which TNF-α utilizes Fn-14 in the synthesis of inflammatory and tissue-destructive mediators in human RASFs and other cells relevant to TNF-α-driven diseases such as IBD and psoriasis. In specific aim 2, we plan to decipher the role of Fn-14 in TNF-α induced function and phenotypic changes in RASFs by using complex multicellular ex-vivo systems, including synovium-on-a-chip model and RA synovial tissue explants. Finally, in specific aim 3, we plan to validate the in vivo efficacy and inhibitory potential of Fn-14- targeted approaches in acute and chronic models of TNF-α-inflammation. The success of these studies will elucidate the role of Fn-14 as a non-canonical signaling receptor of the TNF-α signaling pathway and validate Fn-14 blockade as a potential therapeutic approach for the treatment of RA.

NIDDK - National Institute of Diabetes and Digestive and Kidney Diseases

Project Summary/Abstract In vertebrates, self-renewing hematopoietic stem cells (HSCs) are produced from a developmental event called endothelial-to-hematopoietic transition (EHT). EHT consists of a cellular and transcriptional reprogramming that allows hemogenic endothelial cells (HECs) from a subset of embryonic arteries to leave the vessel and become blood stem cells. HSCs have the capacity to replace and restore the complete blood system upon transplant, making HSC transplant the only curative therapy available for blood diseases like leukemia and lymphoma. Given this therapeutic need, great effort has focused on the development of in vitro protocols that attempt to recapitulate the conditions of EHT for clinical expansion or de novo production of stem cells in the dish. To date none efficiently produce long-lived multipotent HSCs, suggesting that one or more developmental signals for this process remain to be defined. Mechanical forces from blood flow are an essential cue for HSC production via EHT, and the zebrafish Danio rerio provides an excellent animal model in which to study this contribution to hematopoiesis due to conserved molecular genetics of EHT in this species and the ability to observe live embryos with active circulation. Flow-driven EHT is mediated in part by the Yes-associated protein (YAP) transcription factor (TF), a transcriptional coregulator that has roles in organ growth, nutrient regulation and cell fate specification. YAP can be directed to the nucleus as a direct result of physical forces acting on the cell, but the molecular mechanisms by which this promotes EHT and HSC production are unclear. In preliminary data generated under K01 support, single-cell transcriptional analysis of wildtype, yap -/- and YAP-overexpressing HECs from zebrafish point to a role for YAP in regulating a battery of self-renewal hematopoietic TFs, cell cycling and metabolic processes. In examining these YAP gain- and loss-of-function (GOF/LOF) transcriptomes, gene module scores suggest an impaired glycolysis-to-oxidative phosphorylation rewiring in HECs. Genes related to lipid metabolism are also dysregulated by YAP perturbation and can be identified in ‘no flow’ datasets from mouse models. This R03 application will investigate the role of force-directed lipid metabolism in developmental EHT using zebrafish as a model. We hypothesize that hemodynamic forces alter lipid usage in HE to drive the metabolically intensive process of EHT. In the first aim, an unbiased approach of mass spectrometry-based lipidomic profiling will be used to quantify the abundance of lipid species in wildtype and YAP gain or loss of function (GOF/LOF) whole-embryo and sorted endothelial cell populations to determine those metabolites that are YAP-regulated (as a proxy for a major cellular transducer of mechanical force). In the second aim a candidate pathway, the secreted sphingosine-1-phosphate lipid mediator, will be studied for its role in EHT by live-imaging, chemical perturbation and state-of-the-art genome editing technologies to create tissue-specific LOF zebrafish lines. Findings from this proposal will uncover force-driven metabolic responses that might enhance production of HSCs via EHT and generate critical preliminary data to support R01 applications.

NINDS - National Institute of Neurological Disorders and Stroke

ABSTRACT Colony stimulating factor 1 receptor (CSF1R)-related leukoencephalopathy is a devastating genetic neurodegenerative disease characterized by abnormal glial responses, marked white matter pathology in the brain, and severe motor and cognitive symptoms. Currently, there are no effective therapies against CSF1R- related leukoencephalopathy, and its etiology remains poorly understood. Despite the limited consensus regarding its underlying cellular mediators and mechanisms, CSF1R loss-of-function and deficient microglial function are thought to contribute to the disease pathogenesis. However, little is known about the immune changes at the border tissues of the brain, namely at the meninges, and how changes in meningeal immunity and lymphatic drainage might contribute to the initiation and progression of brain pathology in CSF1R- related leukoencephalopathy. Unexpectedly, our preliminary data shows a decrease of lymphatic vessels in the dura of the established model of CSF1R-related leukoencephalopathy (Csf1r+/- mice) and the Fms-intronic regulatory element (FIRE) knockout mice (Csf1rΔFIRE/ΔFIRE), a new mouse model of CSF1R- related leukoencephalopathy. We hypothesize that a deleterious innate immune response at the brain-meningeal interface might affect the pathophysiology of CSF1R- related leukoencephalopathy by dampening lymphatic drainage of the CNS. In this project, we will use the Csf1rΔFIRE/ΔFIRE mouse model to further explore the changes in the central nervous system (CNS) associated immune cells, namely in meningeal myeloid cells, and its association with impaired lymphatic drainage of the brain. I will determine the cellular mechanisms and immune mediators linked to the absence of meningeal lymphatic vasculature in Csf1rΔFIRE/ΔFIRE mice (Aim 1). Secondly, I will use different experimental approaches to enhance meningeal lymphatic drainage in Csf1rΔFIRE/ΔFIRE mice and determine its therapeutic impact on exacerbated glial activation, white matter degeneration, and behavior deficits (Aim 2). This proposal will establish whether impaired meningeal lymphatic vasculature represents a previously unappreciated pathological phenomenon, and a potential therapeutic target, in CSF1R-related leukoencephalopathy.

NHLBI - National Heart Lung and Blood Institute

PROJECT SUMMARY Cardiac development is a complex process that occurs early during embryonic development and requires precise regulation. Early cardiac development, especially of the endocardial and myocardial lineages, is implicated in the development of congenital heart diseases (CHDs). Unfortunately, despite CHDs being the most frequent birth defect affecting approximately one in 100 live births, very few mechanisms are understood about many different forms of CHDs. Proper cardiac development requires precise post-transcriptional regulation, such as alternative splicing, translation initiation, and mRNA modification. These functions are mediated by RNA-binding proteins (RBPs), and unsurprisingly, mutations to many RBPs are implicated in cardiac development and formation of CHDs. One such RBP is DDX3X, which is known from clinical evidence to have implications in CHDs: patients with mutations in DDX3X exhibit DDX3X syndrome, marked by neurodevelopmental disorders and increased risk of CHDs. However, DDX3X has not been well-studied to date in the context of heart development. My proposed research will study the dosage-sensitive effects and mechanisms of DDX3X in the endocardial lineage during cardiac development. In Aim 1 of this proposal, I will determine the effects of reduced DDX3X levels on endocardium formation and function. Using transgenic mice, I will conditionally delete DDX3X in male and female mouse endocardium. Expression level of DDX3X in the endocardium will be quantified using immunofluorescent imaging and flow cytometry. I will then characterize resulting phenotypes by examining phenotypic onset, structural and functional consequences, and cellular and molecular consequences in the heart using a combination of brightfield and immunofluorescent imaging, weight measurements, and echocardiograms. In Aim 2 of this proposal, I will identify the molecular mechanisms of DDX3X in the endocardium. Using immunofluorescent imaging, I will characterize the subcellular localization of DDX3X in the endocardium. I will then identify the direct regulatory network of DDX3X in the endocardial lineage using RNA-seq and Ribo-seq. Targets will be validated against pre-existing eCLIP data as well as in an in vitro system of human pluripotent stem cell-derived endocardial cells. The results of this research will contribute to our understanding about RNA biology, gene dosage, development and disease.