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Camptothecin analogs as "block and lock" agents for HIV

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NIAID - National Institute of Allergy and Infectious Diseases

Project summary/abstract Despite effective antiretroviral therapy (ART), people with HIV (PWH) continue to have chronic inflammation and comorbidities driven by low-level viral transcription from integrated HIV proviruses. Silencing this residual HIV activity could reduce immune activation and improve long-term health. Our long-term goal is to develop therapies that suppress HIV expression and inflammation in PWH on ART. Topotecan (TPT), a Camptothecin analog that inhibits Topoisomerase I, potently suppresses HIV transcription in latently infected T cells. Notably, TPT appears to inhibit HIV independent of its Topoisomerase I activity, suggesting an alternative mechanism of action. We will evaluate new Camptothecin analogs as HIV “block- and-lock” agents. Our central hypothesis is that these compounds can stably suppress HIV without harming host cells. We will pursue three aims: 1) Determine the mechanisms by which TPT inhibits HIV gene expression; 2) Identify new Camptothecin analogs with HIV inhibitory function; 3) Determine the longevity of Camptothecin analog-induced HIV suppression and validate their function using samples from PWH ex vivo. First, we will define how TPT blocks HIV by mapping epigenetic changes at the viral promoter (via CUT&RUN), testing Tat dependence, and assessing post-transcriptional effects like RNA stability and nuclear export (Aim 1). Second, we will screen Camptothecin analogs—with and without Topoisomerase I activity—to identify compounds that suppress HIV at low doses without cytotoxicity. Lead candidates will be validated in primary cells, and their mechanisms and off-target effects will be characterized (Aim 2). Third, we will test whether these compounds can durably silence HIV in latency models and in cells from PWH ex vivo (Aim 3). Completion of these studies will clarify how Camptothecin analogs suppress HIV and assess their therapeutic potential. We expect this work will enable the development of novel “block-and-lock” drugs that reduce persistent inflammation and improve health outcomes in PWH on ART.

Up to $719K
2031-01-31
health research

Free to search & build · $99 one-time to unlock the application pack · No subscription

Cell Type Specific Drug Repurposing for Alzheimer's Disease

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NIA - National Institute on Aging

ABSTRACT This proposal presents an innovative and interdisciplinary approach to accelerate the development of effective, precision therapeutics for Alzheimer’s disease (AD), a complex and heterogeneous neurodegenerative disorder with limited treatment options. Recognizing the critical role of neuroimmune and vascular dysfunction in AD pathogenesis—particularly within the perivascular spaces, the we will leverage state-of-the-art single-nucleus transcriptomic profiling (VINE-seq) and integrative computational drug repositioning to identify and test repurposed compounds capable of modulating disease-relevant gene expression signatures at the cell type level. The project builds on extensive preliminary data identifying key AD-related transcriptional signatures across neurons, glia, and vascular/perivascular cells, revealing both known and novel therapeutic targets. Using publicly available and lab-generated transcriptomic datasets, the team employs tools like Connectivity Map and LINCS to predict drug candidates that reverse pathogenic gene networks in distinct brain cell types. Early analyses have already nominated over 80 potential compounds, including letrozole, irinotecan, sirolimus, and vorinostat, several of which show multi-compartment activity and strong preclinical promise. These candidates will be rigorously validated in vitro using iPSC-derived neurons, glia, and assembloid models, and in vivo using mouse models of AD with tools like longitudinal bioluminescent imaging and plasma biomarkers to assess efficacy, target engagement, and safety. Aim 1 seeks to validate and optimize repurposed drugs targeting neuron-glial dysfunction by integrating computational predictions with functional studies in co-culture systems, chronic mouse dosing models (e.g., 5xFAD, hTAU), and multi-omic readouts of parenchymal pathology, including amyloid burden, neuroinflammation, and synaptic loss. Aim 2 focuses on the vascular-perivascular axis of AD by using single-cell signatures from VINE-seq to identify and test compounds that reverse vascular dysfunction and immune dysregulation in CAA-associated AD models (e.g., TgAPP23), with endpoints including cerebral amyloid angiopathy, blood-brain barrier integrity, and perivascular inflammation. Together, these complementary aims form a robust platform to evaluate and prioritize sex- and cell-type-specific therapies that address both neuronal and vascular drivers of AD. The collaboration brings together a world-class team with expertise in AD genomics, drug repurposing, systems immunology, mouse modeling, and human cell-based models. By uniting cutting-edge transcriptomic technologies with in vivo pharmacology and a precision medicine framework, this work has the potential to deliver impactful therapeutic strategies that can be rapidly translated into clinical trials for patients with Alzheimer’s disease.

Up to $1.4M
2031-05-31
health research

Free to search & build · $99 one-time to unlock the application pack · No subscription

Cell-type specific evaluation of globus pallidus pars externa as a translational target to improve sleep and arousal

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NIH

Significance to VA. Disrupted sleep occurs in many conditions common in Veterans, including VA research priorities such as post-traumatic stress disorder (PTSD), substance abuse, major depression and neurodegenerative disorders. Insomnia is associated with an increased risk for suicide and increases the risk for relapse in opioid use disorder. Furthermore, disrupted sleep and abnormal sleep spindles are common in expensive, hard-to-treat conditions such as dementia and schizophrenia. Increasing evidence suggests that sleep spindles are needed for memory consolidation. Deep sleep is vital in clearing the toxic proteins which are increased by traumatic brain injury and cause neurodegeneration. Accordingly, novel strategies to promote deep restorative sleep are needed for a wide variety of disorders affecting Veterans. Cognitive treatments have utility but exert a relatively modest effect and the most widely used existing pharmacological treatments may disrupt deep restorative sleep and be habit-forming. Thus, alternative treatments are needed. Innovation and Impact. Here, in mice we evaluate for the first time whether deep restorative sleep can be enhanced through cell-type specific modulation of an important basal ganglia structure, the globus pallidus, pars externa (GPe). Our data suggest inhibition of GPe neurons which express the calcium-binding protein parvalbumin (PV+) shortens sleep latency and strongly increases the depth and consolidation of sleep. Our novel data show that another group of neurons which express the transcription factor neuronal PAS domain 1 (Npas1+) regulate sleep spindles, important in memory consolidation. Thus, pharmacological or brain stimulation strategies which inhibit GPe PV+ neurons may be a novel strategy to treat insomnia, whereas inhibition of Npas1+ neurons may promote memory consolidation. Our new data in Aim3 identify a safe natural pharmacological agent which opens potassium channels to inhibit GPe PV+ neurons and promote sleep. Specific Aims. Here, experiments focus on two major, non-overlapping neuronal-types of the GPe, PV+ and Npas1+, which make up 50 % and 30 % of GPe neurons, respectively. Aims 1 and 2 will use state-of-the- art neuromodulatory approaches in genetically-modified mice to test the effects of exciting or inhibiting these neurons on sleep. Aims 1 and 2 will also extend our approach to wild-type mice and identify the downstream neuronal targets. In Aim 3 we will test whether we can enhance sleep via systemic or local application of a safe natural compound which preferentially inhibits GPe PV+ neurons by opening potassium channels. Methodology. All aims investigate the effects of GPe manipulations on sleep and cortical electrical oscillations using electroencephalographic and electromyogram recordings in mice. Aim 1 uses chemogenetics to excite or inhibit GPe PV+ or NPas1+ neurons. Chemogenetics allows a prolonged increase or decrease in neuronal activity through the cell-type specific expression of G-protein activated receptors which respond selectively to an otherwise inert drug. Aim 2 will use closed-loop optogenetics to specifically inhibit (Aim2a) or excite (Aim2b) PV+ or Npas1+ during non-rapid-eye-movement sleep. Optogenetics allows fast and precise manipulation by applying light to neurons expressing light-activated ion channels or pumps. Aim 3 will use a pharmacological approach to inhibit GPe PV+ neurons by opening the potassium channels they express. Path to translation/implementation: Potentially the fastest translational application of this research is to use a pharmacological approach, as in Aim3, to enhance sleep by inhibiting GPe PV+ neurons via opening of the potassium channels they express. Several potassium channel openers, including the natural one we test here are safe and approved for use in humans and proposed as therapeutic agents. Another approach could be to modify existing electrical deep brain stimulation protocols or non-invasive stimulation approaches to target GPe to promote healthy sleep and daytime alertness. Ultimately, in the longer-term, we believe that cell- type specific approaches such as optogenetics or chemogenetics could be applied.

2030-03-31
health research

Free to search & build · $99 one-time to unlock the application pack · No subscription

CELLENONE X1 NEO SYSTEM FOR PROTEOMICS RESEARCH

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OD - NIH Office of the Director

PROJECT SUMMARY This application requests funds to purchase a cellenONE X1 Neo single-cell isolation and liquid dispenser system from Cellenion. The proposed instrument will be located in the Mass Spectrometry Proteomics Core at Baylor College of Medicine. The cellenONE X1 Neo is essential for establishing ultra-low-input proteomics capabilities at BCM and will be primarily used to isolate single cells and subcellular structures from clinical samples, pre-clinical patient-derived xenograft models, and various animal- and cell-based model specimens. Configured with protein sample processing workflows for bottom-up mass spectrometry, this platform will meet critical sample preparation needs for picogram- and nanogram-level starting materials, which demand extremely precise, low-volume, and contamination-free sample handling not adequately supported by our standard core protocols. Single-cell and spatial proteomics is a novel and rapidly advancing area of biomedical research with the potential to transform our understanding of cellular heterogeneity and disease mechanisms. However, despite BCM’s extensive research infrastructure, this capability is currently lacking at our institution. The addition of the cellenONE X1 Neo represents a significant leap forward, enabling our core to offer a complete, automated solution for ultra-sensitive proteomic analysis. Importantly, this instrument will leverage a recent acquisition of the Bruker timsTOF Ultra2 in the Mass Spectrometry Core – a $1.2 million investment in state-of-the-art mass spectrometry instrumentation capable of measuring ultra-small-scale proteomes. Although this Bruker timsTOF is already used for other challenging applications, the absence of a suitable single-cell preparation platform remains a critical barrier to adopting true single-cell and spatial proteomics workflows. The unifying aim of the projects supported by the cellenONE X1 Neo is to explore the molecular mechanisms driving normal physiology and disease at the level of individual cells or rare cell populations. This instrument will provide the precision, scalability, and operational robustness necessary to meet the evolving needs of our growing user base. Its integration into the Mass Spectrometry Proteomics Core aligns with BCM’s strategic plan to expand and share cutting-edge proteomics capabilities and will directly enhance research initiatives in cancer, metabolic disease, neuroscience, immunology, and beyond. The cellenONE X1 Neo will position the Core, and BCM more broadly, as a regional leader in ultra-sensitive proteomics by providing a comprehensive, accessible sample processing solution.

Up to $360K
2027-04-30
health research

Free to search & build · $99 one-time to unlock the application pack · No subscription

Cellular and molecular mechanisms of IL-17A in the pathogenesis of psoriatic arthritis

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NIAMS - National Institute of Arthritis and Musculoskeletal and Skin Diseases

Project Summary/Abstract Psoriatic arthritis is a chronic and progressive inflammatory arthritis closely linked with psoriasis. Blockade of the IL-17 signaling pathway shows impressive efficacy and excellent safety for treatment of psoriasis but less so for PsA. Tissue penetrance limits the bioavailability of monoclonal antibodies to the joint and fibrocartilaginous enthesis and IL-17A signaling is diversified by local immune cells in tissues rendering IL-17A inhibition less effective. A higher dosage poses the risks of serious infections and certain types of cancer as all IL-17 inhibitors are immunosuppressive therefore to improve clinical outcomes in PsA more selective targeting is required. Herein, we have developed a novel animal model of IL-17A gene transfer where mice develop joint and skin inflammation associated with enthesitis, fingernail psoriasis and onycholysis hallmarks of PsA pathology. We will interrogate our murine model to define the early cellular and molecular pathways that dictate IL-17A-induced pathologies using state-of-the-art transgenic mice and molecular tools. Our work will uncover the pathogenic mechanisms of IL-17A and uncover novel molecular targets that can be exploited for therapeutic intervention and directly benefit PsA patients worldwide.

Up to $568K
2030-03-31
health research

Free to search & build · $99 one-time to unlock the application pack · No subscription

Cellular Basis of Novel Organelle Integration by a Photosynthetic Animal

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NIGMS - National Institute of General Medical Sciences

PROJECT SUMMARY / ABSTRACT Endosymbiotic events have driven the evolution of dramatic and consequential biological traits, however most known examples occurred billions of years ago, resulting in deeply integrated associations that mask early steps in the evolution of these associations. “Solar powered, sap sucking” Sacoglossan sea slugs represent a recent and incompletely established endosymbiosis in which stolen, functional chloroplasts are stored in slug tissues. Here, I propose to leverage this unique organism to understand the mechanistic basis of stolen chloroplast (kleptoplast) retention and integration into host cell physiology as a model for understanding endosymbiosis more broadly. To accomplish this, I will: (Aim 1) use organellar proteomics to identify proteins critical for establishment and maintenance of this endosymbiosis (often called “functional kleptoplasty”), (Aim 2) determine the function of those proteins using a combination of pharmacology, patch clamp experiments, and heterologous expression studies, and finally (Aim 3) expand these experiments in a comparative approach using different species of Sacoglossan slugs with different retention abilities. Thus, this study will employ rigorous methodology combining state-of-the-art sequencing technology and multi-omics approaches with physiological approaches, functional assays, and a diverse set of imaging techniques. Further, I will leverage the incredible biodiversity of sea slugs to empower a comparative approach for a comprehensive understanding of a truly unique biological phenomenon: the maintenance of functional photosynthetic chloroplasts within animal tissues. This project will take place at host institution Harvard Medical School (HMS, Cell Biology) and builds on my expertise in transcriptomics, evolutionary biology, and bioinformatics with training in new skills from my primary sponsor, Dr. Corey Allard (HMS, Cell Biology), in electrophysiology, biochemistry, and molecular techniques. I will be aided by an interdisciplinary advisory team composed of my co-sponsor, Dr. Wade Harper (HSM, Cell Biology), an expert in organelle proteomics, Dr. Steven Gygi (HMS, Cell Biology), a world leader in proteomics, and Dr. Amy Lee (HMS, Cell Biology/Dana Farber Cancer Institute), an expert in transcription and RNA biology. Investigation of functional kleptoplasty in Sacoglossan sea slugs will fill a critical gap in our understanding of the evolution of endosymbiosis and the integration of novel organelles into host cells. Completion of these aims will reveal mechanisms used by slugs for sequestration and maintenance of chloroplasts, and thus advance our understanding of the origins and functions of organelles and the fundamental processes that drive the evolution of novel traits. These insights may have broad applications in biotechnology, and could serve as a blueprint to engineer other types of cells to perform photosynthesis, or to take up different foreign cargoes for applications in agriculture, medicine, or even space travel.

Up to $77K
2028-11-30
health research

Free to search & build · $99 one-time to unlock the application pack · No subscription

Cellular Responses to Environmentally Driven Replication Stress

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NIEHS - National Institute of Environmental Health Sciences

PROJECT SUMMARY Numerous environmental agents (hereafter, genotoxins) induce DNA lesions or create other types of barriers that stall replication forks. This can lead to replication stress, and failure to alleviate this stress and restart stalled forks can cause genome instability. Elevated replication stress and the replication-associated mutations that result from genotoxin-induced fork stalling contribute to aging, inflammation, cancer, and numerous other chronic diseases in humans. This project aims to advance understanding of the cellular responses to replication stress. One crucial aspect of the replication stress response involves replication fork reversal, a process that remod- els both the nascent and parental DNA strands to form a four-way junction structure. Fork reversal is carried out by a family of ATP-dependent translocases. Why multiple enzymes with similar activities are involved in this process is not understood. We showed that one of these translocases, HLTF, prevents a remarkably DNA dam- age-tolerant mode of replication by promoting fork reversal and preventing alternative and potentially error-prone modes of DNA synthesis. The unanticipated resilience of the replication fork to various genotoxins in HLTF’s absence may drive mutagenesis and promote the survival of damaged cells. We will investigate the cellular responses to genotoxic and oxidative damage, focusing on the mechanism of replication fork reversal and the impact of loss of fork reversal on genome stability and cell fitness. By combining molecular, biochemical, genomic and proteomic approaches, as well as state-of-the-art single-molecule ap- proaches, we will address the following broad questions: What are the functions of fork reversal and how does it occur in response to environmentally relevant forms of genotoxic damage? What are the specific functions of a central regulator of fork reversal, HLTF, in the face of oxidative and genotoxic DNA damage? Does loss of HLTF and fork reversal increase mutation (rates) in the context of environmental stressors? Our strategy will initially focus in large part on HLTF, elucidating its unique roles. As the project evolves, we will phase in further studies on other remodelers so as to understand, over the long-term, the overall fork reversal process and the unique contributions of each protein to the replication stress response. The knowledge we gain from this research will ultimately facilitate the development of new strategies that i) alleviate the pathological states observed in the absence of the replication stress response, and ii) prevent cancer cells' ability to tolerate DNA damage and develop drug resistance.

Up to $700K
2034-03-31
health research

Free to search & build · $99 one-time to unlock the application pack · No subscription

Center for Advanced Molecular Pathogenesis (CAMP)

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NIGMS - National Institute of General Medical Sciences

Montana State University (MSU) is the alma mater of Maurice Hilleman, the most prolific vaccine scientist of the 20th century. The Hilleman legacy is important to MSU and a point of pride in the state of Montana. Beyond the Hilleman Scholars program and the Hilleman Research Symposium, MSU prioritizes “consequential” biomedical research aimed at understanding the molecular basis of disease. This research is supported by a BSL-3 (select agent capable) laboratory, a large animal BSL-2 facility, the only contemporary cryo-electron microscope capable of high-resolution structure determination the region, state of the art confocal and chemical imaging labs, and new investments in a cell analysis core. This existing infrastructure helps support the long-term goal of this Phase 1 Center of Biomedical Research Excellence (COBRE) to establish a sustainable Center for Advanced Molecular Pathogenesis (CAMP) that fosters recruitment, development, and retention of investigators who share a common interest in translating new discoveries in pathogenesis into new treatments, vaccines, and cures. The theme of this application is intended to be broad enough to cultivate and maintain a pipeline of new faculty with diverse model systems, techniques, and scientific perspectives, while maintaining a unified research goal that focuses on understanding mechanisms and developing treatments for disease. Significantly, this application takes advantage of a talented pool of COBRE-eligible investigators in three departments from two Montana campuses and anticipates the recruitment of additional cohorts of early-stage investigators that will benefit from professional mentoring, instrumentation, and infrastructure essential to advancing biomedical excellence at Montana State University. To achieve this long-term goal, we propose three specific aims: 1) Recruit, develop, and retain a critical mass of investigators necessary to sustain a rigorous, productive, and merit-based research Center focused on Advance Molecular Pathogenesis (CAMP), 2) Improve capacities to translate basic research discoveries into tangible benefits for individuals and society, and 3) Establish core facilities that advance the aims of early-stage investigators, and simultaneously expands the biomedical research infrastructure in Montana and the surrounding IDeA-eligible states. To accomplish these aims, we propose three initial research projects, supported by two cores: The Administrative Core and the Advanced Biological Imaging and Cell Analysis Core. The rigor and feasibility of this application is enhanced by the leadership team (which includes two former COBRE investigators), the prior training and track record of our Research Project Leaders, tailormade mentorship teams, the research infrastructure supported by this award, and active engagement with world-renowned members of our advisory committee. Success of the current cohort will create a syphon, pulling new investigators into a high-priority career development program, thereby building a critical mass of investigators with the necessary momentum to sustain a unified research Center that focuses on understanding mechanisms and developing treatments for disease.

Up to $2.1M
2030-12-31
health research

Free to search & build · $99 one-time to unlock the application pack · No subscription

Center for Nanoscale Science and Technology Postdoctoral Researcher and Visiting Fellow Measurement Science and Engineering Program

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National Institute of Standards and Technology

The National Institute of Standards and Technology (NIST) Center for Nanoscale Science and Technology (CNST) is establishing a financial assistance program for awardees to develop and implement with the CNST a Postdoctoral Researcher and Visiting Fellow Measurement Science and Engineering Program. This program is intended to promote research, training, and practical experience in nanoscale science and technology on-site at the CNST, and to advance the CNST s mission to support the development of nanotechnology through research on measurement and fabrication methods, standards and technology, and by operating a state-of-the-art nanofabrication facility, the NanoFab. The primary program objectives are: 1.To advance, through cooperative efforts with one or more universities, research consistent with the mission of NIST, and CNST specifically. See http://www.nist.gov/cnst/ and 15 U.S.C. Sec. 271 et seq. 2.To provide training for the next generation of nanotechnologists by providing recent Ph.D. recipients postdoctoral positions ( Postdoctoral Researchers ) to perform research at the CNST under the mentorship of a CNST Project Leader. The Postdoctoral Researchers must show promise as contributors to the mission of the CNST, and be selected on the basis of ability and of the relevance of the proposed work to the mission of the CNST. 3.To provide advanced training and access to the CNST s expertise and instrumentation by providing practicing scientists and engineers in the public and private sectors visiting senior research positions ( Visiting Fellows ) to perform research at the CNST in collaboration with a CNST Project Leader. The Visiting Fellows must be selected on the basis of ability and on the relevance of the proposed work to the mission of the CNST. 4.To provide Postdoctoral Researchers and Visiting Fellows under this program with professional development opportunities, including travel to relevant workshops and conferences. 5.To encourage U.S. industrial, university, and government scientists to participate in research at the CNST, either in collaboration with the CNST research program or by using the NanoFab, by providing support for travel and local expenses for participants traveling beyond a normal commuting distance to the CNST in Gaithersburg, Maryland.

rolling
sciencetechnology

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