Search Grants — Free, No Account Required

NIDDK - National Institute of Diabetes and Digestive and Kidney Diseases

PROJECT SUMMARY The kidney is responsible for maintaining an overall state of homeostasis, which it achieves by filtering a large volume of plasma through millions of epithelial units termed nephrons. It is estimated that 1 in 7 adults in the US suffer from chronic kidney disease (CKD), resulting from impairment of these functions. A seminal event in the development of every nephron is the formation of a connection with a collecting duct that establishes a patent pathway for the drainage of fluid into the collecting system and eventually out of the kidney. Human kidney organoids derived from pluripotent stem cells (hPSCs) are an innovative and powerful tool to study renal pathophysiology and to potentially inform new therapeutic avenues, but they suffer from several major limitations. One paramount deficiency is the lack of collecting ducts and failure of their nephrons to drain into a collecting system. To address this problem, we have recently developed a novel platform in which nephrogenic mesenchyme is combined with ureteric bud (UB) progenitor cells to produce an integrated system of collecting ducts within the organoids. Like in vivo development, the nephrons connect with the UB through a controlled fusion event with the distal GATA3+ segment that establishes a continuous passage for fluid flow. Currently, nephron connectivity in this system is limited by a relatively low efficiency of distal specification, and we hypothesize that finetuning the molecular processes underlying fusion will enable more complete integration of nephrons with collecting systems. In this proposal, we will apply this integrated organoid model to address major gaps in our mechanistic understanding of distal nephron patterning and fusion. In Aim 1, we will investigate how WNT and NOTCH signaling interactions coordinate and synergize to control nephron segmentation, and these mechanisms will be applied to enhance nephron-UB fusion in the organoid model. Aim 2 will delineate the cell- autonomous mechanisms underlying distal nephron fusion. Preliminary data show that expression of GATA3 is sufficient to induce fusion-competent distal tubules. Here we will build on this understanding by dissecting the genetic mechanisms downstream of GATA3 and how they regulate this unique cell biological property. Collectively, this work will generate novel and important insights into the mechanisms underlying nephron fusion, and simultaneously it will lead to further advancement in our state-of-the-art organoid model by informing methods to optimize nephron-UB connectivity. We anticipate that these approaches will yield seminal improvements in the structural organization of hPSC-derived kidney organoids, which will translate to progress in the functional capabilities of these tissues and their utility as models for investigation of renal pathophysiology and potentially for developing new therapies for CKD.

NIDDK - National Institute of Diabetes and Digestive and Kidney Diseases

Project Summary Therapies for insulin-dependent diabetes remain inadequate. Islet cell transplantation using donor islets or stem cell-derived tissue could revolutionize treatment, but preventing immune rejection of the transplanted beta cells remains a critical unsolved issue. Current approaches focus on editing hypoimmune islets or redirecting T regulatory suppressor cells, two exciting but challenging strategies. As a novel and more flexible approach, we recently developed synthetic suppressor T cells that use engineered synNotch receptors to sense local disease signals and, in response, induce the targeted expression and delivery of customized immune suppressive payloads. These synthetic suppressor cells are capable of locally targeted immune suppression, without systemic suppression. Our goal is to design optimal therapeutic suppressor cells that can protect transplanted islets by locally blocking immune rejection and promoting tolerance. The flexibility of this platform allows us to systematically explore what payloads and other cell modifications result in the most effective and durable immune protective behaviors. Our aims are: Aim 1. Systematically screen combinatorial suppressive payloads to identify those that yield synthetic suppressor cells that can effectively block key components of cell-mediated rejection (CD4+, CD8+, CD4+Th17, NK, B) and humoral mediated rejection (e.g. complement). Combinations will then be tested for ability to protect islets in several in vivo immune rejection models. Programs will be triggered by synNotch receptors that detect antigens specifically on the beta cell surface, including targets like GLP1R. Aim 2. Introduce modifications to synthetic suppressor cells to increase their survival and persistence, including versions of growth and survival cytokines designed to function in a fully autocrine manner (i.e. cause synthetic suppressor cells to survive without inducing proliferation/inflammation via other immune cells). These survival signals will be induced in a local manner via Islet-sensing synNotch receptors. Suppressor cells with these modifications will be tested for improved persistence in islet immune protection. Aim 3. Engineer synthetic suppressor cells to take up residence in islets by engineering them with chimeric synthetic cell adhesion molecules that recognize islet antigens. Together, these aims should yield optimized suppressor cells that are strongly retained in islets, durably persist only in islets, and produce combinatorial suppressive payloads that prevent immune rejection. This work will yield a better fundamental understanding of immune rejection and the requirements for immune protection and would yield approaches to local immune suppression relevant to solid organ transplantation and autoimmune disease.

NCI - National Cancer Institute

PROJECT SUMMARY/ABSTRACT The advent of genome editing provides the opportunity to treat genetic diseases at their root cause. However, clinical translation remains limited by the challenge of delivering genome editors efficiently and specifically to target cells in vivo. Delivery vehicles are needed to protect genome editors, engage cell-surface receptors and release active enzymes into target cells. Inspired by viral tropism, we developed Enveloped Delivery Vehicles (EDVs), lentivirally derived lipid vesicles that encapsulate CRISPR–Cas9 ribonucleoproteins and display fusogens or engineered antibody fragments on their surface. EDVs enable receptor targeting, but how antibody fragment density and binding affinity control uptake, biodistribution and genome editing remain poorly defined. Building on our laboratory’s expertise with EDVs and CRISPR-Cas9 mechanistic biology, our goal is to define how antibody fragment presentation on EDVs influences their ability to target hematopoietic stem cells (HSCs), a clinically important cell population for treating blood and immune disorders. HSCs are a compelling target for blood cancers, because myeloid malignancies such as acute myeloid leukemia and myelodysplastic syndromes originate from malignant HSCs. Our central hypothesis is that an optimal range of ligand density and affinity (i.e., avidity) maximizes selective uptake and genome editing in HSCs while minimizing off-target uptake by bystander cells. We will test this hypothesis through two aims: 1) Quantify how EDV avidity affects uptake and genome editing in human HSCs ex vivo. 2) Quantify how EDV avidity affects biodistribution and genome editing specificity in humanized mouse models. Mouse models are indispensable for delivery vehicle development because invertebrate, in vitro, organoid, and computational systems cannot recapitulate the immune, spleen, liver, and bone marrow environments or the biophysics of blood flow and tissue perfusion that govern the biodistribution and elimination of delivery vehicles. Unlike prior approaches that focused solely on maximizing antibody affinity or surface presentation, this proposal systematically dissects how density and affinity interact to determine avidity, uptake, and editing efficiency. Completion of this project will produce new targeted delivery vehicles for HSC editing and establish quantitative rules for tuning delivery vehicle avidity to maximize potency and specificity. These results will be broadly applicable to other delivery platforms and accelerate the development of safe and effective in vivo genome editing therapies.

NEI - National Eye Institute

Project Summary Corneal diseases pose a significant public health challenge in the United States, often leading to vision impairment and decreased quality of life. Mesenchymal stem cell (MSC) delivery to the cornea after a severe injury has shown promise by accelerating repair and significantly suppressing inflammation. However, a major bottleneck in developing MSC therapy for corneal repair is the lack of effective delivery methods. Moreover, optimizing the dosage and timing of MSC therapy is crucial for achieving therapeutic outcomes while minimizing side effects. MSCs must also survive and integrate into corneal tissue to exert their therapeutic effects. To date, MSCs have been delivered via surface injection, fibrin gel, or as a sheet on an amniotic membrane. However, these methods are limited by poor MSC survival and/or rapid matrix degradation. To address these issues, we propose the development of adhesive hydrogels that can effectively encapsulate and release MSCs in a sustained manner while having similar biomechanics as the corneal tissue. Our platform composed of a single hybrid polymeric structure with tunable variables to generate two distinct mechanical properties and degradation rates: 1) a soft/controlled degradable adhesive hydrogel to function as a bandage containing MSCs that release secreted factors for promoting corneal epithelial regeneration and 2) a strong/highly adhesive hydrogel that can adhere to corneal stromal defects and simultaneously serves as a stromal replacement while providing a platform for the delivery of MSCs to promote repair of stromal injuries/ulcerations. Our proposed biomaterial is a photocurable adhesive composite hydrogel based on chemically modified gelatin and hyaluronic acid (HA), encapsulated with MSCs. First, gelatin will be dual-functionalized with methacrylic anhydride (MA) and phenylboronic acid (PBA) to control mechanical properties and promote tissue adhesion. The incorporation of methacrylate HA derivatives in the hydrogel will also control the viscosity of the prepolymer and improve its mechanical properties. The physical properties of the resulting hydrogels, such as stiffness, swelling ratio, and degradation rate, which affect MSCs differentiation, will be tuned by varying polymer ratios, degree of polymer functionalization, final polymer concentration, and crosslinking time. We will first optimize the mechanical properties of the proposed hydrogels, and their degradation rates will be tuned to achieve a rate supporting MSCs growth and proliferation (Aim 1). We will then assess the in vitro epithelial proliferation and MSCs differentiation using in vitro models developed in our labs (Aim 2). Finally, we will test the in vivo efficacy of MSC-laden hydrogels using two animal models: a corneal epithelial wound healing model and a corneal stromal injury model (Aim 3). Based on our preliminary data, we anticipate that successfully achieving the Specific Aims of this project will result in a novel treatment that enhances MSC survival and retention by providing a 3D environment resembling the corneal extracellular matrix. The treatment is expected to improve visual outcomes, seal and repair stromal injuries, facilitate re-epithelialization, and reducing the healthcare system burden.

NCI - National Cancer Institute

Abstract This project focuses on macromolecular prodrug-based delivery of a topoisomerase I inhibitor, SN22, structurally enhanced to overcome tumor defense mechanisms in order to achieve durable suppression of high-risk neuroblastoma (NB), the most common and deadly solid tumor of childhood. The intensive, multimodality treatment currently used clinically fails in over half of high-risk NB patients: 50-60% experience a relapse with no curative salvage treatment options. Centered on developing and optimizing a drug delivery strategy against the aggressive disease not responding to conventional therapies, with a particular focus on a high-risk form of multiple drug-resistant NB with increased “stemness” driven by a MYCN protooncogene and its downstream target, ABCG2 (an ABC drug efflux pump suppressing chemosensitivity and promoting tumorigenicity), this project will evaluate an approach integrating polymer-linked prodrug design and structural optimization of the cargo to improve delivery, extend drug residence in the tumor, and reverse drug resistance. Guided by our past work and the results of our preliminary studies toward this project, we hypothesize that prodrug-mediated delivery of SN22 will potently suppress growth of aggressive, pre-therapy and chemorelapsed NB tumors by enhancing drug uptake and extending tumor exposure to therapeutically effective drug levels and by taking advantage of the inactivation-resistant molecular design of this agent. This hypothesis will be tested by pursuing the following specific aims: Aim 1 studies will focus on in vitro evaluation of a series of prodrug constructs on NB cells derived at relapse from MYCN-amplified high-risk NB tumors; Aim 2 studies will examine tumor uptake, biodistribution and elimination of the prodrugs in orthotopic xenograft NB models; Aim 3 experiments will comparatively evaluate antitumor efficacy of prodrug- mediated delivery in models of newly diagnosed and recurrent MYCN-amplified NB in comparison to a new syngenetic model of disseminated (MYCN-driven) high-risk disease. Through optimizing the design and performance of SN22 prodrugs using a panel of clinically relevant models recapitulating distinct types and phases of aggressive NB, this research is expected to have a strong impact on the field by addressing several barriers to the translation and clinical implementation of macromolecular prodrugs and by paving the way to improved clinical management of drug-resistant NB and other high- risk cancers showing minimal or no response to conventional therapies and currently lacking effective treatment options.

NIAID - National Institute of Allergy and Infectious Diseases

ABSTRACT Glioblastoma (GBM) is the most common and aggressive primary brain tumor in adults, with a median survival of only 15–18 months even with therapy. The poor prognosis reflects the tumor's rapid growth, cellular heterogeneity, and resistance to current treatments. Oncolytic virotherapy has emerged as a promising adjunct, and Zika virus (ZIKV) has unique potential given its natural tropism for glioblastoma stem-like cells. While attenuated ZIKV strains have shown preclinical efficacy in reducing tumor burden, challenges remain regarding safety, incomplete tumor clearance, and limited control over viral replication. To address these limitations, we propose to develop a novel optogenetic gene circuit, BiViSTA (Bioluminescent Viral Sensor and Transcriptional Activator), that detects ZIKV infection and couples it to therapeutic gene expression. BiViSTA leverages a split- luciferase sensor that reconstitutes upon ZIKV NS5 protein binding, producing bioluminescence that activates a light-sensitive transcription factor to drive pro-apoptotic or immunogenic transgenes. This approach uniquely links viral infection to targeted tumor cell destruction, creating a self-amplifying and tumor-restricted therapeutic system. We will pursue three Specific Aims: Aim 1 will engineer and optimize BiViSTA constructs responsive to ZIKV NS5 and validate their function in permissive human cell lines. Aim 2 will integrate BiViSTA into 2-D and 3-D GBM culture models to test selective induction of pro-apoptotic genes and assess enhanced ZIKV-mediated killing. Aim 3 will evaluate tumor-specific activation and therapeutic efficacy in an orthotopic xenograft mouse model of GBM, including rigorous safety assessments. Our multidisciplinary team combines expertise in synthetic biology, neuro-oncology, and viral pathogenesis, supported by strong preliminary data demonstrating a “plug and play” optogenetic system. The proposed work is highly innovative as no existing technology dynamically couples virus infection to therapeutic gene expression in real time. Successful completion will open the door to a transformative strategy that will improve the safety and efficacy of oncolytic virotherapy for GBM that can integrate nonpathogenic ZIKV pseudovirions. Beyond GBM, BiViSTA represents a broadly adaptable platform for treating virus-associated cancers and for dynamically controlling gene expression in basic and translational research.

NIDDK - National Institute of Diabetes and Digestive and Kidney Diseases

Project Summary/Abstract The small intestine, central to nutrient absorption like amino acids and lipids, houses highly responsive Lgr5+ intestinal stem cells (ISCs) in the crypt bottom. Over the past decade, our group and others have utilized the mouse intestine to investigate how dietary interventions (fasting, high fat diet, mitochondria pyruvate shuttle inhibition, and high cholesterol diet) impact ISC fate decisions. Although much focus has been on ISCs, the small intestine is a complex environment that includes a variety of non-epithelial cells including resident immune cells that coordinate ISC function and maintenance. In particular, tissue-resident immune cells produce and secrete the cytokine interleukin-22 (IL-22), which is known to be a critical regulator of epithelial homeostasis. Amino acids constitute many nutrients in various foods. However, little is known about how specific amino acids impact ISC proliferation and intestinal immune-stem cell interactions. My research has uncovered that the amino acid cysteine controls ISC function through two mechanisms: 1) by directly activating PPAR-CPT1A-HMGCS2 mediated ketogenesis in ISCs via mTORC1 suppression, and 2) by indirectly boosting IL-22 production by CD8β+ T cells through activation of epithelial Coenzyme A (CoA) biosynthesis. The aims of my proposal focus on elucidating both stem cell-intrinsic and extrinsic mechanisms by which cysteine enhances ISC-mediated repair after injury. Specifically, I plan to: 1) determine how cysteine regulates ISC self-renewal and differentiation in intestinal homeostasis and injury through the control of ketogenesis; 2) determine the cysteine metabolic pathways that contributes to ISC mediated repair after injury; 3) determine how CD8β+ T cells mediate the cysteine response in ISC-mediated repair after injury. This career development K99/R00 award will be essential to my training and provide significant support as I transition to an independent investigator. It will protect me to receive the comprehensive education and training through the robust MIT/Harvard system, coupled with the exceptional research resources, fruitful partnerships will uniquely position me to embark on an unparalleled journey as a rising independent investigator in stem cell metabolism research.

NIAID - National Institute of Allergy and Infectious Diseases

ABSTRACT Tuberculosis (TB) remains a leading cause of mortality in people living with HIV (PLHIV), with 161,000 deaths in 2023 despite widespread use of combinatorial antiretroviral therapy (cART). Although cART effectively suppresses HIV replication, it does not eliminate the markedly elevated risk of Mycobacterium tuberculosis (Mtb) reactivation in co-infected individuals. This persistent susceptibility is thought to stem from incomplete immune reconstitution, including impaired TH1/TH17 balance, reduced CD4⁺ effector memory T (TEM) cells, and ongoing immune activation in the lung. Our preliminary studies in an established Mtb/SIV rhesus macaque model demonstrate that cART alone fails to restore IL-21 and STAT1 signaling, both critical for immune control of TB and HIV. IL-21, a pleiotropic cytokine produced by CD4⁺ T cells, regulates TH1 and TH17 responses and supports macrophage function. Prior work shows that IL-21-IgFc therapy in SIV-infected macaques is safe and preserves mucosal immunity. We propose that IL-21-IgFc, when administered during early cART, can enhance immune reconstitution, reduce viral persistence, and improve TB control. The central hypothesis of this study is that adjunctive IL-21-IgFc therapy will restore key immune pathways in the lung, limit viral reservoirs, and improve macrophage function—thereby preventing Mtb reactivation in SIV- infected macaques. To test this, we will pursue two specific aims: • Aim 1 will determine how IL-21-IgFc therapy during early cART impacts pulmonary immune reconstitution and Mtb-specific responses. We will assess T cell subsets, STAT1/STAT3 signaling, granuloma integrity, and transcriptional signatures using flow cytometry, scRNAseq, and spatial transcriptomics. Additionally, we will perform cross-species transcriptomic comparisons between NHP and human PBMCs from cART-treated and untreated cohorts to identify conserved immune signatures and pathways associated with protection or disease progression. • Aim 2 will evaluate the effect of IL-21-IgFc therapy on viral persistence and macrophage function. We will measure SIV reservoirs in lung tissue, quantify macrophage proliferation, and assess tissue pathology to determine the impact of IL-21 on HIV-associated innate immune dysfunction. Impact: These studies will elucidate mechanisms by which IL-21 enhances lung immunity and limits pathogen persistence in TB/HIV co-infection. Findings will guide development of IL-21-based immunotherapies, a promising host-directed strategy to improve outcomes in high-burden populations where TB and HIV remain syndemic.

NIBIB - National Institute of Biomedical Imaging and Bioengineering

Abstract Laboratory automation is reshaping scientific discovery by enabling high-throughput, precise, and scalable experimentation. Yet, unlike fields such as manufacturing, transportation, and agriculture—where robotics has driven rapid advances in innovation—laboratory automation has lagged behind. Most commercial lab robots are still built to mimic human pipetting rather than enabling experiments that were previously impossible to execute manually. Closed, proprietary platforms prevent researchers from fully leveraging their computational skills and severely limit training opportunities for students and early-stage investigators. This project addresses those gaps by expanding PyLabRobot (PLR), a free, open-source, Python-based framework for designing, simulating, and executing complex liquid handling protocols across a wide range of robotic systems. PLR is software- and hardware-agnostic, supports complete virtual simulation, and can be run in the cloud, making it accessible to anyone with basic Python skills—no expensive equipment required. By enabling users to develop and debug protocols entirely in simulation, PLR allows students, educators, and researchers to build automation expertise before ever stepping into a physical lab. This significantly lowers the entry barrier and ensures that trainees enter the workforce ready to contribute immediately to automation-driven projects. Python’s ubiquity across STEM and engineering makes PLR a natural training tool for modern science. In a recent graduate-level class piloting PLR in full simulation mode, students with no prior robotics background programmed highly sophisticated workflows running from CRISPRi screening, AlphaLISA and TR-FRET assays, small molecule synthesis, FISH staining, and cell line manufacturing—demonstrating both the accessibility and real-world relevance of the platform. These examples highlight PLR’s potential not just to replicate existing workflows, but to unlock new experimental designs that would be infeasible with manual techniques. This R03 proposal supports three focused aims: (1) Create a web-based repository of validated, sharable protocols and documentation to enable reproducibility, benchmarking, and AI-driven method development; (2) Integrate PLR into cloud platforms like Google Colab and develop a dedicated IDE for real-time visualization, debugging, and seamless protocol execution; (3) Build a digital twin simulation engine that models labware, deck layouts, and liquid handling to support predictive optimization before execution. This project is supported by letters of support from the entire PyLabRobot development team and multiple industry stakeholders, including leading automation companies. By removing the barriers that have long constrained laboratory robotics programming, this work will accelerate discovery, lower costs, and build a highly skilled U.S. workforce ready to lead in bioengineering, automation, and advanced manufacturing.

NICHD - Eunice Kennedy Shriver National Institute of Child Health and Human Development

PROJECT SUMMARY Enteric neuropathies are serious conditions caused by abnormalities in the enteric nervous system (ENS), an extensive neural network that spans the entire gastrointestinal tract and controls gut functions. Reduction in number of neuronal nitric oxide synthesis (nNOS) expressing enteric neurons is a hallmark of many human gastrointestinal diseases, including esophageal achalasia, gastroparesis, and Chagas disease. Despite the prevalence of these conditions and the morbidity they cause, current treatment options are significantly limited and do not directly address the underlying pathophysiology. Cell therapy represents a novel, curative approach to alleviating enteric neuropathies by replacing the absent or injured neurons. Although we have achieved significant successes in our preclinical cell therapy studies using animal models of enteric neuropathy, an enteric neural cell therapy approach specifically tailored for nNOS-deficient diseases would be a significant advance. Recent studies, and our own preliminary experiments, have demonstrated that enteric neural stem cells (ENSCs) generate two neuronal classes that subsequently differentiate into specific phenotypes and that this process includes phenotype switching. This led us to hypothesize that manipulation of this enteric neurochemical plasticity (ENCP) would allow us to optimize ENSC therapy by generating therapeutically relevant, disease- specific neuron subtypes. By utilizing optimized ENSC culture conditions, cell transplantation techniques, transcriptomic analysis, and genetic manipulation, we propose to test our hypothesis that enteric neuronal subtype plasticity occurs postnatally and can be leveraged for cell therapy application. We will also elucidate the role of the transcription factors, Pbx3 and Tbx3 in postnatal ENCP. The proposed studies will advance our understanding of the mechanisms underlying cell commitment in the ENS and how this knowledge could be leveraged to optimize the therapeutic efficacy of cell therapy applications in the future.

National Aeronautics and Space Administration

Awards will be made as cooperative agreements to accredited Minority Serving Institutions (MSIs) partnered with non-profit organizations in the United States that are eligible to apply for this NASA Research Announcement (NRA). The period of performance for an award is up to 2 years. Prospective proposers are requested to submit any questions in writing to NASAMUSIC@nasaprs.com no later than 10 business days before the proposal due date so that NASA will have sufficient time to respond. Proposers to this NRA are required to have the following, no later than the due date: 1) a Data Universal Numbering System (DUNS) number, 2) a valid registration with the System for Award Management (SAM) [formerly known as the Central Contractor Registry (CCR)], 3) a valid Commercial And Government Entity (CAGE) Code, 4) a valid registration with NASA Solicitation and Proposal Integrated Review and Evaluation System (NSPIRES) (this also applies to any entities proposed for sub-awards or subcontracts.) Consult Appendix H Section H.3.1 for more eligibility information. Consult Appendix H Section 2.2 regarding teaming requirements and partnership guidelines. The goal of NASA MUSIC is: to provide strategic effort that will leverage research and contract relationships of MSIs and NASA through relationships developed by non-profit organizations that may include collaboration of subject matter experts and access to NASA research facilities; An effort to improve STEM education and research at MSIs; A funded activity that seeks to build institutional capacity of MSIs; An activity to support long-term sustainability of STEM research at MSIs. MUSIC seeks to address the agency goals and objectives through: Increasing the institutional awareness of NASA competitive resources that can build the capacity of MSIs to offer and conduct STEM undergraduate and graduate research with a focus on NASA opportunities. Assembling MSIs and their stakeholders with common interests, and challenges then provide common tools for MSIs to increase efficiency and optimize resources including opportunities to develop formal and informal partnerships. Connecting MSI administrators and university STEM leaders to cutting-edge initiatives at NASA that can increase interest in securing research and contracting opportunities while supporting NASA s policy to achieve an Agency-wide goal of providing one percent of total contract value of prime and subcontracting awards to MSIs. https://www.hq.nasa.gov/office/procurement/regs/1826.htm To achieve these goals, MUSIC seeks to increase university program capacity about practical uses of research to drive institution sustainability through the following targets: Advance the understanding of MSIs on how to effectively develop institutional administrative support by competing at the university level for funding opportunities, which will result in successful application to, and management of these funding opportunities (including those at NASA). Extend MSI s capabilities by: A. Leveraging the MSIs research capabilities with NASA research to develop Small Business Innovation Research (SBIR) and Small Business Technology Transfer (STTR) projects that develop and demonstrate innovative technologies that fulfill NASA needs and have significant potential for successful commercialization. B. Increasing the preparation of undergraduate and graduate science, technology, engineering, and mathematics faculty with opportunities to participate with NASA researchers and missions through grants and contracts. To achieve these goals and objectives, NASA solicits proposals from MSIs to implement the NASA MUSIC; to engage MSIs in authentic STEM experiences related to NASA missions; and to inspire and captivate learners utilizing NASA s unique assets to develop a keen interest in STEM. Every institution that intends to submit a proposal to this NRA, including the proposed prime award or any partner whether an education institution, other non-profit institutions, and other organizations that will serve as sub-awardees or contractors, must be registered in NSPIRES. Electronic submission of proposals is required by the due date and must be submitted by an authorized official of the proposing organization. Such registration must identify the authorized organizational representative(s) who will submit the electronic proposal. All principal investigators and other participants (e.g. co-investigators) must be registered in NSPIRES regardless of submission system. Potential proposers and proposing organizations are urged to access the system(s) well in advance of the proposal due date(s) of interest to familiarize themselves with its structure and enter the requested information. Electronic proposals may be submitted via the NASA proposal data system NSPIRES or via Grants.gov. Organizations that intend to submit proposals via Grants.gov must be registered 1) with Grants.gov and 2) with NSPIRES. Additional programmatic information for this NRA may develop before the proposal due date. If so, such information will be added as a Frequently Asked Question (FAQ) or formal amendment to this NRA and posted on http://nspires.nasaprs.com. It is the proposer s responsibility to regularly check NSPIRES for updates to this NRA.

NHLBI - National Heart Lung and Blood Institute

PROJECT SUMMARY Lung possesses a remarkable ability to regenerate and restore normal lung function after severe acute injury. However, in the case of chronic injuries, the lung epithelial stem/progenitors fail to restore normal lung function and contribute to a pathologic regeneration, amounting to regenerative failure that typifies chronic restrictive lung diseases like Idiopathic Pulmonary Fibrosis (IPF). Both airway and alveolar epithelial stem cells are capable of differentiating into type 1 and type 2 alveolar epithelial cells (AT1 and AT2, respectively) via a series of intermediate cell states. During normal injury resolution, like in acute injuries in human lungs and murine models, both airway and alveolar epithelial stem cells go through a euplastic intermediate cell (EIC) state to eventually differentiate into alveolar epithelial cells (AT1 and AT2s). However, in chronic lung diseases like IPF or animal models of chronic fibrotic injury, the stem cells progress through EICs and differentiate further into aberrant basal-like dysplastic intermediate cell (DIC) state that result maladaptive regeneration and honeycomb-like cystic structures. Currently, we lack understanding of the signaling mechanisms that control the balance between euplastic and dysplastic regenerative cell states. This is especially relevant in a histologically heterogeneous disease like IPF where the balance between euplastic and dysplastic repair is a key indicator of disease severity. To this end, we discovered that EICs and DICs are spatially distributed in relation to severity of remodeling and state of microenvironment niche signals. EICs are often found in areas of active but mild injury and are associated with mesenchymal cells that secrete pro-regenerative growth factors such as NRG1 that activated ErbB3/STAT3 signaling. On the other hand, DICs are associated with Collagenhigh/CTHRC1high pro-fibrotic mesenchymal cells found in actively remodeling regions. Furthermore, our preliminary data suggests that intermediate filaments such as cytokeratin 17 (KRT17) and Vimentin play an integral role in the switch from EIC to DIC. Therefore, we will test the hypothesis that spatially restricted microenvironment cues activation of distinct signaling pathways in activated epithelial progenitors to determine euplastic versus dysplastic regenerative trajectories in the following three aims: Aim 1: Define the role of NRG1/ErbB3 signaling in euplastic alveolar regeneration. Aim 2: Determine the role of TGFβ1/KRT17 signaling axis in dysplastic alveolar regeneration during chronic injury. Aim 3: Identify spatial cues required for euplastic to dysplastic regenerative switch. We will use newly established animal models of chronic injury, cell-specific in vivo conditional knockout of relevant candidate genes, in vitro co- culture techniques modeling human euplastic and dysplastic repair to test specific signaling components of NRG1/ErbB3 signaling and TGFβ1/KRT17 signaling. Finally, using a novel in silico approach, we will build an integrated mouse PF and human IPF spatiotemporal signaling map of progressive fibrosis. Together, successful completion of this proposal will build a comprehensive model of niche-influenced signaling crosstalk governing epithelial plasticity and reveal novel therapeutic targets for treating fibrotic disorders.

NIAID - National Institute of Allergy and Infectious Diseases

Project Summary HIV cure has been achieved almost exclusively in individuals undergoing allogeneic hematopoietic stem cell transplantation (HSCT) for hematologic malignancies using homozygous donors for the CCR5D32 deletion. Although these cases demonstrate that installation of an HIV-refractory immune system is feasible, broader clinical translation is limited by viral rebound from residual reservoir cells, outgrowth of viruses using alternative co-receptors, and the substantial morbidity associated with allogeneic transplantation. These limitations highlight the need for approaches that both protect newly generated immune cells from infection and actively eliminate persistent HIV-infected cells. We propose autologous HSCT-based strategies that integrate precise genome editing with targeted cellular immunotherapy. Using base editors, we introduce functionally silent missense mutations in CD4, the primary HIV entry receptor, to generate immune cells broadly resistant to both CCR5- dependent and CCR5-independent HIV strains while preserving CD4-dependent immune function. In parallel, we engineer additional CD4 variants that render base-edited cells resistant to CD4-specific CAR T cell-mediated depletion, thereby enabling CAR T cells to selectively eliminate residual wild-type CD4+ cells, including those harboring HIV reservoirs, following HSCT. These approaches establish a protected immune compartment while permitting targeted eradication of reservoir cells that drive viral rebound after antiretroviral therapy interruption. We will test the central hypothesis that combining autologous HSCT with CD4-targeted base editing will both prevent infection by HIV variants with distinct co-receptor tropisms and enable CAR T cells to eliminate persistent infected cells, thereby sustaining viral control while preserving immune competence Aim 1: Develop a CD4-modified HSCT approach to prevent HIV infection while preserving CD4+ immune reconstitution. Aim 2: Develop a CD4-modified HSCT approach enabling CAR T cell elimination of residual HIV-infected cells while preserving CD4+ immune reconstitution.

U.S. National Science Foundation

In alignment with the CREST Program goals, through this solicitation, the CREST Program seeks to expand its geographic diversity and reach by explicitly calling for proposals led by and for efforts in Established Program to Stimulate Competitive Research (EPSCoR) jurisdictions. The U.S. National Science Foundation's EPSCoR program pursues a mission to enhance the research competitiveness of targeted jurisdictions (state, territory or commonwealth) by strengthening science, technology, engineering and mathematics (STEM) capacity and capability through a diverse portfolio of investments from talent development to local infrastructure. For a list of EPSCoR jurisdictions visit https://new.nsf.gov/funding/initiatives/epscor/epscor-criteria-eligibility. EPSCoR CREST Center awards provide support to enhance the research capabilities of institutions through the establishment of centers that effectively integrate education and researchin EPSCoR jurisdictions. EPSCoR CREST Center awards promote the development of new knowledge, enhancements of the research productivity of individual faculty, and an expanded presence of students fromEPSCoR jurisdictionsin science, technology, engineering, and mathematics (STEM) disciplines. Successful EPSCoR CREST Center proposals will demonstrate a clear vision and integration of STEM research and education and will align with the mission of the Division of Equity for Excellence in STEM (EES) with respect to the development of a STEM workforce. EPSCoR CREST Centers are also expected to provide leadership by meaningfully involving the effortsofall individualsinSTEMat all levels. Centers are required to use evidence-based and innovative strategies to address workforce development issues, such as recruitment, retention, and mentorship of participantsfromEPSCoR jurisdictions. Successful proposals are expected to achieve national research competitiveness, broaden participation in STEM, and generate sustained, non-CREST funding from Federal, state, and/or private-sector sources. PhaseIandPhaseIIEPSCoR CRESTCenterAwards Both Phase I and Phase II EPSCoR CREST Center awards provide multi-year support for institutions that demonstrate a strong research base. Phase I EPSCoR CREST Center awards provide funding for five years of research on a specific NSF-supported topic.Institutions may submit a Phase II EPSCoR CREST Center proposal requesting funding to continue research in the same disciplinary area as the Phase I EPSCoR Center or may submit a Phase I proposalfocusedonadisciplinaryareathatis significantlydifferentfromthoseofthepreviousaward(s). EPSCoR CRESTPartnershipSupplements EPSCoR CREST Partnership Supplement requests are invited from current EPSCoR CREST Center recipients. Supplements support the establishment or strengthening of partnerships and collaborations with active CREST Centers and other nationally or internationally recognized research centers (including NSF-supported research centers), private sector research laboratories, K-12 schools, and/or informal science entities, including museums and science centers, as appropriate. Such partnerships and collaborations should aid EPSCoR CREST Centers quest in advancing knowledge and education on a research theme of national significance.

U.S. National Science Foundation

The Established Program to Stimulate Competitive Research is designed to fulfill the mandate of the National Science Foundation (NSF) to promote scientific progress nationwide. NSF EPSCoR facilitates the establishment of partnerships among academic institutions, government, industry, and non-profit sectors that are designed to promote sustainable improvements in a jurisdiction's research infrastructure, Research and Development (R&D) capacity, and R&D competitiveness of EPSCoR-eligible jurisdictions (i.e., states, territories, and commonwealths). Eligibility to participate in the EPSCoR funding opportunities, including the EPSCoR RII: EPSCoR Research Fellows program, is described on theNSF EPSCoR website. EPSCoR RII: EPSCoR Research Fellows directly aligns with the NSF EPSCoR strategic goal of establishing sustainable Science, Technology, Engineering, and Mathematics (STEM) professional development pathways that advance workforce development and effects engagement in STEM at national and global levels. EPSCoR RII: EPSCoR Research Fellows provides awards to build researchcapacityin institutions and transform the career trajectories of investigators and further develop their individual research potential through collaborations with investigators from the nation s premier private, governmental, or academic research institutions and/or centers. The fellowship provides opportunities to establish strong collaborations through extended or periodic collaborative visits to a selected host site.Through collaborative research activities with the host site, Fellows will be able to learn new techniques, develop new collaborations, advance existing partnerships, benefit from access to unique equipment and facilities, and/or shift their research toward potentially transformative new directions. The experiences gained through the fellowships are intended to have lasting impacts that will enhance the Fellows research trajectories well beyond the award period. The benefits to the Fellows are also expected to improve the research capacity of their institutions and jurisdictions more broadly. EPSCoR Research Infrastructure Improvement (RII): EPSCoR Research Fellows offers the following two tracks: 1)EPSCoR Research Fellows: NSF; and 2) EPSCoR Research Fellows: @NASA While the two tracks have similar goals, EPSCoR Research Fellows: NSF is open to a broad community and EPSCoR Research Fellows: @NASA supports faculty from eligible institutions (See Section"IV. Eligibility Information" for more details) to collaborate with researchers at the National Aeronautics and Space Administration (NASA) research centers. PIs who are eligible for both tracks may apply for only one track per competition cycle. Proposals from both tracks are submitted to and merit reviewed by NSF. Awards in the EPSCoR Research Fellows: @NASA track are referred to NASA EPSCoR for distribution of additional NASA funds and other needed NASA coordination required for the award. In both tracks, the EPSCoR RII: EPSCoR Research Fellows program provides opportunities for the participation of one trainee, who must be an undergraduate or graduate student enrolled full-time in an accredited degree program, or a postdoctoral researcher from an EPSCoR jurisdiction. Staff members, such as technicians or lab assistants could be considered as trainees when properly justified.

U.S. National Science Foundation

The Established Program to Stimulate Competitive Research (EPSCoR) supports the U.S. National Science Foundation (NSF) mission by promoting nationwide scientific progress. Through this program, NSF fosters partnerships among academic institutions, government entities, industry, and non-profits. These collaborations aim to drive long-term improvements in research infrastructure, enhance R&D capacity, and boost the research competitiveness of eligible EPSCoR jurisdictions, including states, territories, and commonwealths. A jurisdiction s research ecosystem is the interconnected network of organizations, researchers, trainees, community stakeholders, and resources that contribute to the process of research and innovation that advances fundamental knowledge, generates use-inspired products, and ultimately cultivates beneficial impacts for a jurisdiction. E-CORE supports jurisdictions in building significant and sustainable research capacity and research infrastructure for targeted areas of focus, hereinafter referred to as cores, that underlie a jurisdiction's research ecosystem. Based on the evidence-based and self-identified needs of a jurisdiction, the types of cores supported by E-CORE may include (but are not limited to) development, enhancement, and/or ensuring the sustainability of: research administration; research facilities and infrastructure (including cyberinfrastructure); STEM education (K-12) pathways; higher education pathways; early career investigator pathways; broadening participation; workforce development; national and global partnerships; community engagement and outreach; technology transfer; economic development; and use-inspired research pathways. E-CORE projects must be designed to support the sustainability of the research infrastructure cores beyond the award period. Projects will also support the development and growth of new jurisdiction-wide connections, and the leveraging of existing jurisdiction-wide connections, to drive substantive and sustainable impacts.

U.S. National Science Foundation

The Established Program to Stimulate Competitive Research (EPSCoR) supports the mission of the U.S. National Science Foundation (NSF) by promoting nationwide scientific progress. Through this program, NSF fosters partnerships among academic institutions, government entities, industry, and non-profits. These collaborations aim to drive long-term improvements in research infrastructure, enhance R&D capacity, and boost the research competitiveness of eligible EPSCoR jurisdictions, including states, territories, and commonwealths. A jurisdiction s research ecosystem is the interconnected network of institutions, organizations, researchers, trainees, community stakeholders, and resources that contribute to the process of research and innovation that advances fundamental knowledge, generates use-inspired products, and ultimately cultivates beneficial societal impacts for a jurisdiction. E-RISE supports hypothesis-driven or problem-driven research and fosters the development of research teams and products in a scientific topical area that aligns with a jurisdiction s research ecosystem and priorities, as detailed in the jurisdiction s Science and Technology (S&T) Plan or drawn from other jurisdiction plans, reports, or publications prepared by appropriate authorities or bodies.E-RISE invitesinnovativeproposalswithin the chosen research area thatwillleadto development and implementation of sustainable broad networks of individuals, institutions, and organizations, and that will transform the science, technology, engineering and mathematics (STEM) research capacity and competitiveness in a jurisdiction. E-RISE is particularly interested in proposals that justify exploring emerging or interdisciplinary research areas with high potential impact. E-RISE projects must have a clearly articulated research goal that will lead to new knowledge by addressing a clear hypothesis or problem. The E-RISE projectshould promote (i) areas of research capacity-building within a chosen research topic; (ii) development of a skilled workforce that is relevant to the research topic, as well as the project and its outcomes; (iii) a culture of collaboration and engagement across different types of academic institutions and organizations, as well as non-academic sectors (e.g., industry and government); (iv) integration of the research with societal impacts; and (v) a clear sustainability plan to preserve the resulting research incubator's team and products beyond E-RISE funding.

U.S. National Science Foundation

The Established Program to Stimulate Competitive Research (EPSCoR) is designed to fulfill the mandate of the National Science Foundation (NSF) to promote scientific progress nationwide. EPSCoR eligibility status is yearly updated and reported in the EPSCoR website (see EPSCoR eligibility). Through this program, NSF establishes partnerships with government, higher education, and industry that are designed to affect sustainable improvements in a jurisdiction's research infrastructure, Research and Development (R&D) capacity, and hence, its R&D competitiveness. The FEC program (formerly known as EPSCoR Track-2 program ) builds interjurisdictional collaborative teams of EPSCoR investigators in Science, Technology, Engineering, and Mathematics (STEM) focus areas consistent with the currentNational Science Foundation Strategic Plan. Projects are investigator-driven and must include researchers from at least two EPSCoR eligible jurisdictions with complementary expertise and resources necessary to address challenges, which neither party could address as well or as rapidly independently. FEC projects have a comprehensive and integrated vision to drive discovery and build sustainable STEM capacity that exemplifies institutional, geographic, and disciplinary diversity. The projects STEM research and education activities seek to broaden participation through the strategic inclusion and integration of all individuals, institutions, and sectors. Additionally, EPSCoR recognizes that the development of early-career faculty is critical to sustaining and advancing research capacity.

U.S. National Science Foundation

The Experimental Program to Stimulate Competitive Research (EPSCoR) is a program designed to fulfill the National Science Foundation's (NSF) mandate to promote scientific progress nationwide. The EPSCoR program is directed at jurisdictions that have historically received lesser amounts of NSF Research and Development (R&D) funding. Thirty-one jurisdictions including twenty-eight states, the Commonwealth of Puerto Rico, the U. S. Virgin Islands, and Guam currently are eligible to participate. Through this program, NSF establishes partnerships with government, higher education, and industry that are designed to effect lasting improvements in a state's or region's research infrastructure, R&D capacity and hence, its national R&D competitiveness.Research Infrastructure Improvement Program: Track-2 (RII Track-2) awards provide funds in the range of $1.5 to 2.0 million per year for up to 3 years to consortia of EPSCoR jurisdictions. The awards promote opportunities for collaborations among EPSCoR jurisdictions in all areas of science, engineering, and education supported by the National Science Foundation (NSF). RII Track-2 proposals must describe a clear, comprehensive, and integrated vision to drive discovery, and train a skilled workforce capable of solving science and engineering challenges of regional, thematic, and national relevance. Proposals should also include a strong rationale for the establishment of the consortium and clearly demonstrate that the consortium is well-positioned to produce results that cannot be obtained by any single partner working independently. The Science, Technology, Engineering, and Mathematics (STEM) research and education activities should broaden participation by different types of institutions, individuals, and sectors in the project.

U.S. National Science Foundation

Ethical and Responsible Research (ER2) research projects use fundamental research to produce knowledge about what constitutes or promotes responsible or irresponsible conduct of research and why, as well as how to best instill responsible conduct of research into researchers, practitioners, and educators at all career stages. In some cases, projects will include the development of interventions or applications to ensure ethical and responsible research conduct. The program funds research projects that identify: (1) factors that are effective in the formation of ethical science, technology, engineering, and mathematics (STEM) researchers; (2) approaches to developing those factors in all STEM fields that NSF supports; and (3) why and how those factors and approaches increase responsibly conducted research. Proposals from or involving substantial collaboration with minority-serving institutions, women's colleges, or organizations primarily serving persons with disabilities are strongly encouraged. Proposals that include international collaborations are encouraged if the unique resources, expertise, facilities, or locations of international partners enhance the merit of the proposed work. International partners are required to find non-NSF funding. Please see NSF s PAPPG for further guidance on international collaborations.

NIAID - National Institute of Allergy and Infectious Diseases

Enterovirus D68 (EV-D68) was described in 1962 as a respiratory illness but has recently had increased circulation and disease severity. EV-D68 is associated with acute flaccid myelitis (AFM), paralysis in children characterized by spinal cord lesions. There are no specific treatments for AFM and most children have long-lasting neurological deficits. Neuropathogenesis of EV-D68 is likely a combination of virus- and immune- mediated cytotoxicity, though relevant models to investigate this are limited. We recently showed that neonatal mice paralyzed by EV-D68 have abundant T cell recruitment to the spinal cord. When T cells were depleted, mice were protected from paralysis, suggesting a role for T cells in AFM. The role of T cells in human AFM remains unknown. The absence of human models to study T cell interactions in the central nervous system (CNS) hinders progress in identifying viral targets, mechanisms of neural injury, and immune contribution to pathogenesis. Major histocompatibility complexes (MHC) present antigens to T cells to induce the cytotoxic response and effector and memory T cells. Cell surface MHC Class I (MHC-I) is upregulated on neurons and glial cells after CNS injury. However, MHC-I modulation during viral infection of the CNS has not been studied in a multicellular human model. While viral infections are canonically expected to increase surface MHC-I, many viruses downregulate MHC-I as an immune evasion strategy. Our data suggests that EV-D68 infection downregulates, but does not eliminate, MHC- I in infected hSCO. We developed a human spinal cord organoid (hSCO) model for EV-D68 infection from induced pluripotent stem cells (iPSC). hSCO differentiate into multiple cell types of the spinal cord, including neurons and glial cells, and can be infected by EV-D68. Importantly, hSCO are in suspension without Matrigel, a substance that has hindered incorporation of T cells into CNS organoids due to effects on T cell activation and migration. We will utilize our human organoids to understand MHC-I modulation during EV-D68 infection and to investigate interactions between T cells and EV-D68 infected cells of the human CNS. Our overarching hypothesis is that EV-D68 neuropathogenesis is mediated by effects of CD8+ T cells, which we will test by 1) defining the mechanism of EV-D68 downregulation of MHC-I and 2) examining interactions between T cells and EV-D68 infected hSCO. Results will define mechanisms of EV- D68 neurovirulence in a novel human model by elucidating interactions between infected hSCO and the adaptive immune system. Findings will increase understanding of EV-D68 pathogenesis and potentially identify new virus-specific or immune-specific therapeutic targets for AFM.

NINDS - National Institute of Neurological Disorders and Stroke

Project Abstract: Mitchell Syndrome is a progressive childhood-onset neurodegenerative disorder characterized by sensory ataxia, hearing loss, skin changes, and eventual paralysis and encephalopathy, typically leading to death within the second decade of life. The first patient with Mitchell Syndrome was treated at Washington University in St. Louis (WashU), which makes WashU uniquely situated for researching this rare disease. WashU sees around 30% of known patients, has characterized the disease's natural history, and has extensive resources such as post-mortem tissues and a biofluid biobank. Our long-term goal is to develop effective treatments for Mitchell Syndrome. The disease is caused by an autosomal dominant variant in ACOX1, leading to a gain-of-function in the acyl-CoA oxidase 1 (ACOX1) protein. Our preliminary studies indicate that the variant impacts both sensory neurons and oligodendrocytes. Patient-derived induced pluripotent stem cells (iPSCs) offer a scalable, homogeneous platform for modeling rare diseases and evaluating precision-medicine therapeutics. We aim to develop iPSC-derived models of oligodendrocytes and sensory neurons to recapitulate Mitchell Syndrome as a tool to evaluate potential treatments. Preliminary data from patient-derived iPSC lines show transcriptional and metabolic abnormalities linked to the disease variant. We propose two specific aims: Aim 1: Evaluate ACOX1 gain-of-function in iPSC-derived oligodendrocytes. Aim 2: Investigate ACOX1 gain-of-function in iPSC-derived sensory neurons. Mitchell Syndrome intertwines lipid metabolism, oxidative stress, and neuronal/glial degeneration. This project aims to provide essential models for therapeutic evaluation, leveraging WashU's unique expertise and resources. At the culmination of this project we will have two scalable, disease-relevant, human models of Mitchell Syndrome that can be used for mechanistic studies, therapeutic development, and biomarker exploration – critical steps on the way to treat this lethal and tragic disease.

NCI - National Cancer Institute

PROJECT ABSTRACT The Philadelphia-chromosome negative myeloproliferative neoplasms (MPNs) are clonal hematopoietic stem cell disorders, which include polycythemia vera (PV), essential thrombocytosis (ET), and primary myelofibrosis (PMF). MPNs carry an inherent risk of progression to advanced MPN, consisting of accelerated- phase disease (AP; 10-19% blasts in the peripheral blood or bone marrow), as well as blast phase disease (BP; ≥ 20% blasts in the peripheral blood or bone marrow). The prognosis of patients with advanced MPN remains quite poor, with median survival of 2.6 months. Importantly, chemotherapy regimens used to treat Acute Myeloid Leukemia (AML) such as standard induction chemotherapy (which are often used in advanced MPN) appear to have limited efficacy in this setting. Thus, the treatment of advanced MPN is a major unmet clinical need. We recently carried out a phase I/II study to test the safety and efficacy of combination therapy with the JAK1/2 inhibitor Ruxolitinib and the hypomethylating agent Decitabine in patients with advanced MPN (MPD-RC 109 study; NCT02076191). This combination (RUX-DAC) was based on data demonstrating synergy between these drugs in in vitro preclinical studies. 46 patients were accrued to the phase I and II studies. 37 patients were response evaluable. Complete response (CR) occurred in 10%, Complete Response with incomplete count recovery (CRi) in 24%, Partial Response (PR) in 24%. 42% of patients had no response to therapy. Using samples available from the MPD-RC 109 study, as well as samples from a contemporaneous clinical trial of 28 patients with advanced MPN treated with the RUX-DAC regimen carried out at the MD Anderson Cancer Center (NCT02257138), and samples collected from advanced MPN patients treated with the RUX-DAC regimen as a standard of care at Memorial Sloan Kettering Cancer Center, we seek to assess and validate genetic and epigenetic determinants of response to RUX-DAC in this cohort of homogenously treated advanced MPN patients. Specifically, we seek to assess whether the mutational profile of advanced MPN patients explains and predicts response to therapy. We further seek to assess whether alterations in genomic architecture in advanced MPN occur in patients who respond to therapy. Finally, we seek to determine if the baseline global methylation profile correlates with response to therapy, as has been demonstrated for other myeloid malignancies. Data resulting from these studies could be used to guide therapeutic decisions and identify patients for whom combination RUX-DAC therapy has the highest likelihood of procuring a response, as well as to open new lines of biologic and therapeutic inquiry into this disease.

NHLBI - National Heart Lung and Blood Institute

PROJECT SUMMARY Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) hold promise for cardiac disease modeling, drug testing, medical device testing, and regenerative medicine. However, their limited functional maturity in vitro remains a major barrier to widespread application. We recognize the crucial role of distinct and specific mechanical loads in cardiac morphogenesis and seek to co-opt signals downstream of mechanical engagement to drive maturation of hPSC-CMs. In particular, we seek to establish a causal link between mechanical stimulation and hPSC-CM maturation by focusing on the contribution of extracellular matrix (ECM) proteins, the primary family of proteins that mediate and confer mechanical force to the cell. Our lab has developed a novel human, chambered cardiac muscle pump model (hChaMP) capable of simulating both stretch and shear forces akin to a native cardiac cycle. We have begun to incorporate epicardial-derived cells (namely cardiac fibroblasts, CF) into the hChaMP (termed epi-ChaMP via a previously funded R01), as CFs remodel the ECM in response to mechanical stimulation. We also have expertise in cutting-edge computational modeling approaches to refine mechanical stimulation parameters and in fully characterizing the composition of the ECM following mechanical stimulation. Given our unique ecosystem, we can test the hypothesis that CM maturation is augmented in the epi-hChaMP via potent signaling of an evolving ECM deposited by CF in response to dynamic volumetric pressure. We will do so by developing and validating a computational fluid-structure interaction model that accurately replicates native cardiac pressure profiles in the epi-hChaMP (Aim 1), testing the mechanistic role of ECM deposition and remodeling by FBs in driving cardiomyocyte and tissue-scale maturation under physiologic loading (Aim 2), and by applying a statistical optimization framework to define dynamic volumetric loading regimes that maximize functional maturation of epi-hChaMP tissues. The proposal directly addresses reviewer feedback through clearer model differentiation, enhanced methodological descriptions, and inclusion of a non- cardiac fibroblast control. Completion of this project will reveal unappreciated contributions of ECM to CM maturation (Basic Science Innovation) and will yield a robust in vitro human muscle pump with unprecedented physiological relevance (Applied Science Innovation).