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Camptothecin analogs as "block and lock" agents for HIV

open

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

Canonical and alternative functions of low-density lipoprotein in multiple myeloma

open

NCI - National Cancer Institute

PROJECT SUMMARY Multiple myeloma (MM) remains an incurable malignancy in most patients, who will eventually relapse and become refractory to existing therapies. There is thus a critical need for therapeutic innovations that maintain remission and preserve patient quality of life. The tumor microenvironment (TME) of MM is within hypoxic bone marrow, which is intriguing because membrane biogenesis for proliferation is limited by available cholesterol. The synthesis of cholesterol is energy- and oxygen-intensive, and limited supply leads to resource competition between constantly dividing tumor and hematopoietic cells. This nutrient tug-of-war between tumor and nontumor cells in the MM-TME is evidenced by high rates of anemia and infection in MM patients. Separately, altered immune cell behavior results in immunosuppression, which is common in advanced and relapsing MM, and therapeutically underserved. Epidemiolocal studies indicate that low levels of plasma cholesterol are linked with MM progression, likely reflecting the high demand for sterols in the TME. All cells can rapidly increase cellular sterol levels and stimulate membrane biosynthesis through uptake of cholesterol-rich low-density lipoprotein (LDL). Most recently, it has been published that LDL also transport small RNAs that promote macrophage polarization by activating an endosomal sensor of RNA, toll-like receptor 8 (TLR8). Researchers demonstrated that pharmacologic antagonism of TLR8 shifted the immune landscape within atherosclerotic plaques and reduced disease burden in hyperlipidemic mice. Taken together, LDL is a nutrient-dense particle capable of supporting cell proliferation, and a source of extracellular sRNA capable of modulating immune cell function. The goal of this Stephen I. Katz Early-Stage Investigator Grant is to 1) determine whether LDL’s canonical functions in lipid transport directly enable MM growth, progression and therapeutic resistance, and 2) investigate whether LDL’s transport of small RNAs indirectly enables MM progression through activation of TLR8 in host leukocytes to create an immunosuppressive TME. In agreement with the funding mechanism, this proposal represents an ambitious new direction for our laboratory supported by rigorous work in the literature and an ensemble of experienced collaborators and clinicians that reflect the tremendous environment for translational MM research at our institution. We will harness this translational power by using innovative approaches to humanize lipoprotein metabolism in proven pre-clinical models of MM, and by combining state-of-the-art bioinformatic, imaging, and single-cell immune profiling approaches, to test the therapeutic synergy of safe, effective, and FDA-approved, LDL-lowering drugs with standard-of-care chemotherapy. Mice are used in this study because it is not possible to fully recapitulate the complex patho-physiological state of myeloma disease, which involves multiple cells, tissues and organs, using cultured cell models. The validity of mice as an animal model for studying the pathophysiology and treatment of myeloma disease has been supported by extensive literature showing that cellular and molecular features of myeloma disease in mice are similar to those in humans. Upon completion, this award will fill a critical gap in knowledge of how LDL contributes to a pro-malignant TME, and more specifically how lipoprotein disequilibrium contributes to immunosuppression in MM. We envision that these data will be leveraged to open many new research opportunities for diagnostic and therapeutic approaches for MM, and perhaps, other malignancies.

Up to $402K
2031-07-31
health research

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

Cardiac MRI Phenotyping of Coronary Microvascular Dysfunction

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NHLBI - National Heart Lung and Blood Institute

Project Summary Angina is one of the most common symptoms prompting coronary angiography, yet up to 50% of patients are found to have no significant obstructive coronary artery disease (CAD). Many of these individuals continue to experience chest pain without a clear diagnosis. In a substantial proportion of cases, symptoms are attributable to coronary microvascular dysfunction (MVD), an increasingly recognized contributor to myocardial ischemia, heart failure with preserved ejection fraction (HFpEF), obesity, and diabetes. Despite its high prevalence, MVD remains underdiagnosed and challenging to monitor noninvasively. Invasive coronary function testing (CFT) is the gold standard but is technically complex, carries procedural risk, and is limited to specialized centers. While positron emission tomography (PET) enables non-invasive quantification of myocardial blood flow (MBF) and myocardial perfusion reserve (MPR), its utility is constrained by cost, limited availability, radiation exposure, and moderate spatial resolution. Cardiac magnetic resonance (CMR) offers a promising, widely accessible alternative. Recent advances in quantitative perfusion mapping, Artificial Intelligence-driven image enhancement, and multiparametric tissue characterization have transformed stress CMR into a powerful tool for assessing perfusion and myocardial health with high spatial resolution. These capabilities position CMR to play a central role in the diagnosis and monitoring of MVD. This proposal leverages state-of-the-art CMR technology, cross-modality validation, invasive testing, and mechanistic phenotyping to: • Define and validate CMR-based diagnostic criteria for MVD using invasive CFT as the reference standard. • Evaluate the correlation and comparative diagnostic accuracy of CMR- versus PET-derived MBF and MPR for detecting MVD. • Assess whether GLP-1 receptor agonist (GLP-1RA) therapy improves coronary microvascular function independent of weight loss, using bariatric surgery as a comparator and serial quantitative CMR for longitudinal assessment. This work addresses a critical unmet need for non-invasive diagnostic tools and treatment monitoring in MVD, with the potential to expand precision cardiovascular care for an overlooked yet high-risk population.

Up to $848K
2031-03-31
health research

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

CD4 T cells and immune cell triads in anti-tumor immunity

open

NCI - National Cancer Institute

PROJECT SUMMARY CD8 T cells are powerful adaptive immune cells that have the ability to eradicate cancer cells. However, despite the presence of tumor-specific CD8 T cells (CD8 TST) tumors progress, suggesting that CD8 T cells become dysfunctional. While direct cytotoxic activity against cancer cells generally resides within the CD8 T cell compartment, CD4 T cells play important roles in priming and activation of naïve CD8 T cells in lymphoid tissues and enhancing CD8 T cell mobilization into peripheral tissues, including tumors. We recently investigated if and how tumor-specific CD4 T cells (CD4 TST) could be enlisted to overcome CD8 TST dysfunction/exhaustion in tumors (Espinosa et al., (2024)). Employing preclinical cancer models as well as human samples, we discovered that CD4 TST are critical during the effector phase within tumors: CD4 TST must co-engage with CD8 TST on the same dendritic cell (DC), forming a three-cell-cluster (triad) to license CD8 TST cytotoxicity and tumor destruction. Triad formation reprograms CD8 TST, preventing and even reversing CD8 TST dysfunction/exhaustion, ultimately leading to the elimination of large established tumors and long-term protection from recurrence. We showed that when triad formation is prevented in settings of immunotherapies, tumors progress even when equal numbers of CD8 and CD4 TST are present within tumors. Strikingly, we found that the presence of triads in tumors of patients treated with immune checkpoint blockade was associated with clinical responses. Thus, the unique spatial positioning and cell-cell interactions of CD4 T cells, CD8 T cells, and DC are critical for anti-tumor responses. These intriguing findings have now led us to ask how CD4 TST in immune cell triads license CD8 TST to kill cancer cells. Which transcription factors (TF) and polarization state(s) are required for CD4 TST effector function within triads? Which receptor/ligand and/or cytokine/chemokine signals mediate CD8 TST cytotoxicity? Which signals do DC provide to CD4 TST and/or CD8 TST? Are specialized DC subsets, phenotypes, or maturation states needed? In this application, we will leverage the power of preclinical cancer models and unique human samples, employ state-of-the-art technologies, and design and test innovative strategies to identify the critical molecular factors and signals within triads and test how they can be enforced for the treatment of cancer. This work will provide mechanistic insights into the immune cell interactions critical for tumor control and inform strategies to enhance immunotherapy responses.

Up to $671K
2031-06-30
health research

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

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