Search Grants — Free, No Account Required

NIGMS - National Institute of General Medical Sciences

Project Summary Signals that cells receive over time from a small set of pathways (e.g., BMP, Wnt, and TGFβ) shape their fate and phenotype during development, regeneration, and disease. Despite their central importance, signaling histories of individual cells are often inaccessible to direct observation, hindering quantitative analysis and obscuring their connection to eventual cell fate. This challenge is particularly pronounced in mammalian systems, where limited optical access and the constraints of size and timescale often render live imaging impractical. To address this issue, we have developed an approach to reconstruct the history of signaling activity in single cells based on endpoint fluorescence images. This is achieved by regulating CRISPR base editors to generate mutations in engineered target sites at rates proportional to the signal of interest. These mutations create a heritable record of signaling activity in the genome, which can be read out at a later time, together with the gene expression profile of the cells. Using this approach, we demonstrated that cells retain a memory of their past response level to BMP signaling for up to 18 days, providing a mechanism for long-term interactions between signals that can facilitate coordination of developmental processes over time. In this proposal, we will expand the scope and utility of our signal recording approach by extending its dynamic range to capture the broad spectrum of in vivo signal intensities and enabling simultaneous recording of the sequence and timing of two signaling pathways. We will also engineer mouse embryonic stem cells to record three key developmental pathways: BMP, Wnt, and Nodal. This will allow us to generate stem cell-derived embryo models and chimeric embryos to link cell fate and spatial organization at the onset of organogenesis with signaling activity at different time windows earlier in development. Additionally, we will investigate mechanisms that enable long-term changes in BMP responsiveness following an initial stimulation, without requiring differentiation. We will then test whether similar mechanisms exist in Wnt and Nodal pathways and assess their role in mediating long-term crosstalk between pathways. To achieve these goals, we will take an interdisciplinary approach combining gene editing, quantitative imaging, epigenomic assays, computational analysis, and generation of developmental models. The proposed goals build on my prior publications, recent preliminary data from our lab, and collaborations I have established since launching my lab. This research program will substantially advance the state of the art in molecular recording, transforming it into a technology that can be used in vivo, in mammalian systems to drive biological discovery. Our long term vision is to identify how signaling history controls cellular decision making during development, and how instructions that cells receive are coordinated over time to produce tissues with the correct number, types, and spatial arrangement of cells. Ultimately, this knowledge will inform strategies for tissue engineering, and open new avenues for understanding and treating diseases driven by dysregulated signaling.

NIAID - National Institute of Allergy and Infectious Diseases

PROJECT SUMMARY Aging leads to widespread changes in the immune system, including increased inflammation, diminished vaccine responsiveness, and greater susceptibility to infection and chronic disease. However, older adults vary significantly in their immune trajectories—some maintain robust function, while others experience immune decline. The biological mechanisms driving this heterogeneity remain poorly understood, particularly the role of upstream hematopoietic stem and progenitor cells (HSPCs), which continually regenerate the immune system throughout life. This project tests the central hypothesis that epigenetic remodeling of HSPCs contributes to immune-related hallmarks of aging and variability in these phenotypes, shaping both inflammatory set points and immune responsiveness in older adults. To address this, we leverage a well-characterized, longitudinal cohort of 62 older adults, each of whom received a different influenza vaccine over three consecutive seasons. Using our novel PBMC-PIE platform—developed to enrich and profile circulating HSPCs from cryopreserved PBMCs at single-cell resolution—we can interrogate progenitor cell states and their progeny without requiring bone marrow biopsies. This enables unprecedented, longitudinal insight into how human HSPCs are reprogrammed with age and vaccination. In Aim 1, we will identify molecular and epigenomic features in HSPCs and innate immune cells that distinguish strong from poor vaccine responders. In Aim 2, we will determine how different influenza vaccine platforms (high-dose, adjuvanted, recombinant) shape the chromatin landscape and lineage bias of HSPCs and their progeny over time. In Aim 3, we will test whether maladaptive “aged” HSPC states can be rejuvenated in vitro using candidate immunomodulatory factors identified in Aims 1–2. By linking blood-based epigenetic signatures to real-world immune outcomes, this work will reveal fundamental mechanisms of immune aging, identify biomarkers of immune resilience, and inform the development of targeted strategies to restore immune function in older adults.

NIGMS - National Institute of General Medical Sciences

Project Summary The University of Missouri-Columbia (MU) proposes to direct a Trans-disciplinary Basic Biomedical Science (TDS) T32 training program for predoctoral students in PhD programs. The mission of our training program is to facilitate the transformation of our trainees into independent scientists who are active producers of new biomedical knowledge and are adept at solving the complex problems of disease and illness that adversely affect human health. Our training program accomplishes this mission by bringing together four PhD programs including Biochemistry, Biological Sciences, Biological Engineering, and Translational Biosciences. The latter program includes parallel emphasis areas (tracks) covering specialties including microbiology & virology, gene & stem cell therapy, immunology, physiology, nutrition & exercise science, cancer biology, and epidemiology & precision health. We propose to use science identity as our organizing principle, with its composition of community belonging, self-efficacy, and achieved deliverables. Objective #1: To strengthen the safe, supportive, and nurturing nature of our community by providing training to improve the quality of the peer- and mentor-mentee relationships that are so critical to career success and satisfaction. Objective #2: To expedite fluency in the scientific method, we will institute a Critical Thinking Workshop to be followed by a new course, Biomedical Literature Colloquium, for primary literature critique, presentation, and writing. Objective #3: To promote professional communication skills and increase achieved deliverables in the form of external fellowship wins and peer-reviewed publications. Objective #4: To improve the PhD completion rate of trainees in the participating biomedical doctoral programs from the current level of 74-81% to 85%. Our active program evaluation structure will allow independent, professional assessment of trainees’ sense of science self- efficacy, science identity, perceived support, and satisfaction with mentoring relationships, all of which are predictors of academic satisfaction and persistence outcomes among predoctoral students. The TDS T32 program will implement a wide-ranging set of programmatic activities to develop the technical, operational, and professional skills that will enable our trainees to flourish in their independent scientific careers.

NIGMS - National Institute of General Medical Sciences

Project Summary Double-strand breaks (DSBs) in DNA represent one of the most severe threats to genomic integrity, with improper repair leading to mutations, chromosomal translocations, and oncogenic transformation. Cells have evolved robust pathways, including non-homologous end joining (NHEJ), homologous recombination (HR), and microhomology-mediated end joining (MMEJ), to repair DSBs. While much is known about protein-mediated DSB repair mechanisms, the role of transcript RNA in modulating repair remains poorly understood. Yet, the potential contribution of RNA molecules, particularly transcript RNA with sequence complementarity to DNA ends, remains largely unexplored. Recent discoveries from the PI’s laboratory demonstrate that RNA transcripts can bridge broken DNA ends and facilitate repair via NHEJ and MMEJ in both human and yeast systems, suggesting a conserved and fundamental role for RNA in genome maintenance. This project seeks to systematically uncover the mechanisms, regulatory proteins, and biological consequences of transcript RNA involvement in DSB repair across multiple contexts in human cells. Aim 1 establishes isogenic human embryonic kidney (HEK293T) and diploid retinal pigment epithelial (RPE-1) cell lines expressing spliced or non-spliced RNAs from the same adeno-associated virus integration site 1 (AAVS1) safe harbor locus and investigates how these transcripts modulate end-joining repair (NHEJ and MMEJ) of chromosomal DSBs induced by CRISPR/Cas9, followed by deep sequencing and bioinformatics analysis. Aim 2 identifies the key proteins and molecular processes involved in RNA-mediated end-joining repair in human cells. This goal will be achieved by inhibiting core NHEJ and MMEJ factors using chemical inhibitors and siRNAs, and by selectively reducing nascent spliced and non-spliced RNA transcripts using CRISPRi. These perturbations will define how individual proteins and transcript RNAs contribute to the efficiency, fidelity, and pathway choice of RNA-mediated end joining. Aim 3 examines whether transcript RNA differentially modulates DSB repair in exonic and intronic sequences of endogenous human genes in both HEK293T and RPE-1 cells, including breaks induced by CRISPR/Cas9 and mutagens, to understand RNA’s role in maintaining genome integrity. The significance of this work lies in establishing transcript RNA as a previously unrecognized regulator of DSB repair pathways, revealing new dimensions of genome stability control and potential therapeutic targets. The results could provide insights to improve genome editing technologies, inform strategies to reduce mutagenesis and carcinogenesis, and contribute fundamentally to understanding how RNA functions extend beyond traditional regulatory roles to directly safeguarding the genome. The outcomes will also lay groundwork for exploring RNA-mediated DNA repair in stem cells, non-dividing cells, aging models, and disease states, potentially transforming approaches to cancer prevention and therapy.

NCI - National Cancer Institute

Abstract PAX3 is an indispensable upstream regulator of normal melanocyte survival, population expansion, and migration, and these functions are subverted in melanoma proliferation, resistance to apoptosis, and metastasis. Despite significant advances in treatment, melanoma is still a deadly cancer with considerable mortality rates, and PAX3 contributes to this by supporting cancer aggressiveness, resistance to therapies, and relapse. While PAX3 is expressed in developing melanoblasts and melanocyte stem cells, downregulation of PAX3 during terminal differentiation is required. In melanoma, PAX3 expression is overexpressed, maintained, and essential for cancer progression. Melanoma cells rely on PAX3 function; indeed, a loss of PAX3 expression leads to catastrophic loss of melanoma cell growth, migration, and viability. Due to the significance of PAX3 in melanoma, it is surprising how much is unknown about PAX3 function. Classically defined as a transcription factor, only a handful of PAX3 downstream target genes in melanoma are known. Our lab and others have uncovered select downstream effector genes that can explain some but not all of the wide roles that PAX3 has on melanoma survival and progression. To uncover clues on other functions of PAX3, our laboratory performed an unbiased immunoprecipitation and mass spectroscopy (IP/MS) screen to identify PAX3 binding partners. Unexpectedly, we discovered that PAX3 interacted with several proteins involved with RNA surveillance, decay, and splicing. Further, our preliminary data support that this role in RNA regulation is independent of the transcriptional role of PAX3. The goal of this proposal is to uncover how PAX3 regulates these cell functions through downstream effector genes, by acting directly on genetic regulation or indirectly through other molecular means. To address the significant gaps in knowledge into PAX3 function, the hypothesis guiding this proposal is that PAX3 can dynamically act on downstream genes through canonical and noncanonical transcription factor functions, with outcomes modified by binding partners, tumor stage, and small molecule inhibitors. The major scientific impact of this proposal is to reveal a PAX3 dependent genomic enhancer map in melanoma connected to regulated genes, discover mechanisms for modulating this signature, and exploit these pathways as targets for therapy. The innovation of this proposal lies in a more detailed focus on PAX3, in terms of what genes are controlled and how. We propose the innovative hypothesis that PAX3 is not purely a canonical transcription factor and can function both transcriptionally and post-transcriptionally. This work is significant since the PAX3 transcriptional signature in melanoma is still undiscovered, and it is not clear if it is static or adaptive at different tumor stages or drug treatments. This work will uncover PAX3-dependent pathways driving melanoma progression and reveal potential molecular weak points that can be targeted by therapeutics. Use of mice in this proposal: The rationale for utilizing mouse models is that normal and pathological cells behave differently in vivo due to a complex microenvironment that cannot be replicated in other model systems.

NIMH - National Institute of Mental Health

Project Summary Mutations that reduce or alter the activity of repressive chromatin modifiers are frequent causes of neurodevelopmental disorders. The inaccessibility of the developing and the dynamic nature of neurogenesis so far have prevented an adequate understanding of how molecular pathologies arise downstream of the loss of specific chromatin regulators. This knowledge gap represents a hurdle for developing therapeutic interventions for neurodevelopmental disease. This proposal aims to combine targeted protein depletion with highly efficient directed differentiation regimens for human pluripotent stem cells to dissect gene regulatory functions of the autism-associated chromatin repressor EHMT1 during human cortical neurogenesis. Based on the extensive characterization of a novel, multipurpose (degradation/immunoprecipitation/visualization) degron allele, we hypothesize that interactions with cell type-specific co-factors allow EHMT1 to control the expression of stage- specific target genes during neurogenesis, resulting in the accumulation of molecular alterations and cortical neuron (CN) dysfunction when EHMT1 is lost from early development onwards. To systematically test this hypothesis, we will first determine whether molecular alterations caused by EHMT1 deficiency from earlier stages of neurogenesis onwards accumulate in CNs and to what degree dysregulated gene loci and CN function remain responsive to restoring physiological EHMT1 levels (Aim 1). We will then combine genomics, proteomics, and genetic approaches to identify how EHMT1, together with candidate recruiters and co-factors, regulates distinct gene loci at specific stages of cortical neurogenesis (Aim 2). Finally, we will expand our degron approach to dissect the functional interplay of different autism-associated H3K9 methylation regulators during human neurogenesis to identify interactions between these proteins that could be clinically exploited (Aim 3). Our experiments will determine currently unknown gene regulatory functions of disease-associated chromatin repressors at critical stages of human cortical neurogenesis. By generating mechanistic insight into how deficiencies of EHMT1 and other H3K9 methylation regulators introduce molecular pathologies in CNs, we anticipate revealing new opportunities for therapeutic interventions with specific neurodevelopmental diseases.

NEI - National Eye Institute

PROJECT SUMMARY/ABSTRACT Vision loss caused by the death of photoreceptors is a leading cause of irreversible blindness worldwide, yet therapeutic options remain limited. For this reason, the NEI’s Retinal Disease Program has identified the development of strategies for the treatment of retinal degenerations as a core program goal. Recently, several laboratories have derived photoreceptors from stem cells, making cell-replacement therapies particularly promising. Additionally, important advances have been made into manipulations that could stimulate retinal regeneration from the retinal Müller glia. The critical barrier for the success of such therapies is to understand the factors required to direct fate decisions in progenitors towards fully differentiated cell subtypes that are also capable of properly rewiring into retinal circuits. Although key transcription factors have been identified as essential for generating retinal cell classes, the target genes required to generate each retinal cell subtype are still undetermined. TBX2 is a central transcriptional regulator of all photoreceptor subtypes and is highly conserved across vertebrates. Our main hypothesis for this proposal is that retinal progenitors express TBX2 to repress the identity of photoreceptor subtypes that are not UV cones through two different mechanisms, and our main goal is to identify these two roles. Three Specific Aims are proposed: Specific Aim 1 will test the hypothesis that tbx2a and tbx2b have subdivided functions and their roles in generating photoreceptor subtypes are different. Specific Aim 2 will identify candidate factors downstream of TBX2 important for the generation of photoreceptor subtypes. Specific aim 3 will test the hypothesis that differences in Tbx2a and Tbx2b function are caused by differences in the repression domain. At the successful completion of this project, I will have identified the subdivided functions of tbx2a and tbx2b and subsequent targets to generate and maintain photoreceptor subtypes. This proposal benefits from the experimental accessibility of the retina and our deep knowledge of retinal cell types and circuits, but our approach has the potential to impact the study of other neuronal degenerative diseases. In addition to my proposed research, I designed a holistic training plan that will help me advance toward my goal of leading an independent research lab. The work planned for the F31 award period will be valuable in terms of both research and training opportunities.

U.S. National Science Foundation

TSL combines interests and resources of separate programs in the Division of Research on Learning in Formal and Informal Settings (DRL) to explore the opportunities and challenges implied by innovative visions of the future for STEM learning.?? The TSL program invites interdisciplinary teams of STEM content specialists, experts in relevant technologies, STEM formal and informal education specialists, researchers with expertise in the learning sciences, and specialists in education research and evaluation methods to submit proposals for research projects that (1) Study efficacy of existing prototypes for innovations like virtual schools, special STEM schools, and educational programs that combine opportunities of formal and informal learning resources in their communities; or (2) Design and conduct exploratory development of new potentially transformative models for STEM learning environments.?? The cross-cutting proposals will draw from work in the four primary DRL programs: Discovery Research K-12 (DR K-12), Informal Science Education (ISE), Research and Evaluation on Education in Science and Engineering (REESE), and Innovative Technology Experiences for Students and Teachers (ITEST).?? However, proposals submitted in response to this solicitation must have a scope that extends well beyond any of those programs individually.

U.S. National Science Foundation

The Transforming Undergraduate Education in Science, Technology, Engineering, and Mathematics (TUES) program seeks to improve the quality of science, technology, engineering, and mathematics (STEM) education for all undergraduate students. This solicitation especially encourages projects that have the potential to transform undergraduate STEM education, for example, by bringing about widespread adoption of classroom practices that embody understanding of how students learn most effectively. Thus transferability and dissemination are critical aspects for projects developing instructional materials and methods and should be considered throughout the project's lifetime.?? More advanced projects should involve efforts to facilitate adaptation at other sites.The program supports efforts to create, adapt, and disseminate new learning materials and teaching strategies to reflect advances both in STEM disciplines and in what is known about teaching and learning.?? It funds projects that develop faculty expertise, implement educational innovations, assess learning and evaluate innovations, prepare K-12 teachers, or conduct research on STEM teaching and learning. It also supports projects that further the work of the program itself, for example, synthesis and dissemination of findings across the program. The program supports projects representing different stages of development, ranging from small, exploratory investigations to large, comprehensive projects.

U.S. National Science Foundation

This solicitation addresses issues of translation and diffusion that arise in moving knowledge gained from fundamental learning and education research toward application in PreK-12 STEM classroom practice or leveraging knowledge derived from effective practice toward driving fundamental research. The first goal of this funding opportunity is to encourage the scientific study of theories, frameworks, and models for the translation and diffusion of knowledge, especially between fields and across contexts and levels-of-analysis (e.g., biological to cognitive/socioemotional to behavioral; individual to classroom to broader demographic variables; lab to classroom to school to district). The second goal is to advance or move specific practice, research or scientific discovery in STEM education reciprocally along the research-practice continuum. The Translation and Diffusion (TD) solicitation invites four types of proposals: Research on Translation or Diffusion proposals request funding to conduct scholarship that will advance the sciences of translation or diffusion of fundamental research knowledge toward PreK-12 formal STEM education practice by developing or refining theories, frameworks, or models (or adapting those from other domains) and conducting related research. Such proposals may also address the leveraging of effective classroom practices toward the enrichment of foundational research, constructs and models. We note that bi-directional movement across boundaries is a mutually beneficial reciprocal process. Proof-of-Concept Research proposals request funding to explore the feasibility and viability of particular knowledge or products generated from STEM education research toward advancing practice in formal PreK-12 settings (even if it is still basic or applied research and development rather than implementation). The goals are to facilitate the process by which the promise that the initial insight holds for research and practice can be realized. The outcome of such a project would lay the methodological, theoretical, empirical, design, or social foundation for conducting systematic work at the next stage of development or at the next level of analysis. Empirical and theory-building efforts to adapt initial insights from research or practice across significantly different contexts, populations, domains, and levels-of-analysis are also welcome. Synthesis proposals critically integrate the current state of knowledge on a particular topic relevant to translation and diffusion in formal PreK-12 STEM education. Such proposals should include the state of the knowledge across disciplinary communities and across relevant literatures, identify the lacunae in STEM education knowledge, and, where appropriate, lay out thenext steps for future research and development. Conference / Workshop proposals relevant to the call are also welcome.

NHLBI - National Heart Lung and Blood Institute

Project Summary/Abstract Despite surgical advancement and endovascular device innovations, endothelial injury remains a fundamental barrier in vascular surgeries, contributing to post-operative complications such as vein graft failure, restenosis, and thrombosis. Current surgical approaches offer no regenerative solution to actively protect endothelial health during or after interventions. To address this gap, we have developed a bioengineered mitochondrial therapy platform that leverages lipid-polymer-peptide surface functionalization to enable targeted mitochondria delivery, enhanced cellular uptake, and integration into the host mitochondrial network. Mitochondrial dysfunction is a key driver of impaired endothelial repair following vascular surgeries, yet mitochondrial transplantation has not been adopted clinically due to challenges in storage stability, scalability, and compatibility with surgical workflows. Our preliminary studies demonstrate successful surface engineering of stem cell–derived mitochondria with robust uptake and functional rescue in dysfunctional endothelial cells. In vivo, we show that engineered mitochondria were selectively uptake by target vessels during both intra-operative (ex vivo vein graft storage and arterial intraluminal infusion) and post-operative stages (intravenous injection) upon vascular surgeries. To further establish the translational potential of bioengineered mitochondrial therapy, we propose the following studies. Aim 1 will optimize mitochondrial source selection, storage formulation, and surface modification parameters to ensure translational feasibility. Aim 2 will validate the therapeutic efficacy and biodistribution of the optimized system in two highly clinically relevant in vivo models of open (vein graft bypass) and endovascular surgeries (balloon angioplasty). This interdisciplinary project integrates surgical sciences, bioengineering, translational medicine, and device development to establish a next-generation therapeutic approach for vascular surgeries. By advancing a procedural-compatible, device-enabled delivery system, this work directly addresses a critical need in vascular surgery by developing translatable tools and technologies to improve surgical outcomes in millions of patients with cardiovascular diseases.

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

ABSTRACT Transposable elements (TEs) make up over half of the human genome and are increasingly recognized as key regulatory sequences of gene expression in development and disease. While cellular intrinsic transcriptional and epigenetic mechanisms controlling TE sequences are well-documented, whether and how TEs response to external microenvironmental signals, particularly mechanical forces, remains unexplored. This represents a critical gap in understanding both TE regulation and genome–environment interactions. Our preliminary data reveal that several TE families, including LTR7 from the primate-specific HERV-H family, function as Mechano-Response Enhancer Elements (MREEs) that regulate gene expression and human pluripotent stem cell fate in response to mechanical stimuli. Additional preliminary findings support our central hypothesis that TEs act as MREEs by modulating human genes and cell fate of human pluripotent stem cells through a mechanism at least partially governed by the key mechano-effector YAP, which regulates TEs’ local epigenetic activity and facilitates their long-range chromatin looping with target genes in response to mechanical changes. To test this model, we propose three specific aims: 1) determine how mechanical signals regulate the local chromatin activity of TE MREEs; 2) elucidate how mechanical cues mediate long-range chromatin interactions between TE MREEs and their distal target genes; 3) delineate the mechanism by which TE MREE modulating human embryonic stem cell fate. Our objective is to rigorously establish the novel concept that TEs function as MREEs, uncover their underlying molecular mechanisms, and assess their biological significance in regulating developmental genes and hESC fate. The expected outcome will not only shift the current paradigm of mechanobiology from protein- coding genes to non-coding regulatory elements such as TE but also improve our understanding of TE regulation and genome-environment interactions in health and disease.

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

Summary: The negative effects trauma exposure, such as those that promote PTSD, can be passed to offspring. While the environment in which affected parents raise their offspring clearly plays a role, a significant genetic component also exists. However, some of this genetic influence, typically revealed by twin studies, may stem from epigenetic inheritance; as implied by studies in male rodents, where trauma-specific changes in sperm miRNA content can lead to trauma-specific behavioral changes in all offspring. Evidence for epigenetic inheritance in humans is mainly epidemiological, but recent studies have revealed stress-induced sperm miRNA changes consistent with this idea. For example, we found that men with high Adverse Childhood Experiences (ACEs), whose children can be negatively affected, show reduced sperm levels of miR-34/449 family members; mirroring findings in mice exposed to chronic social instability (CSI) stress across generations. In mice, these sperm miRNA changes persist in preimplantation embryos post-fertilization, altering early embryonic gene expression that leads to elevated anxiety and impaired sociability in female offspring as well as reduced levels of sperm miR-34/449 in males. Our new data driving this proposal reveals another example. The degree of men’s exposure to adult trauma, as assessed by the Trauma History Questionnaire (THQ), which measures PTSD risk, correlates with the levels of miRNAs 532, 361, 375, and 491 in their sperm. The highest THQ scores are associated with 4- to 130-fold increases in these levels. In contrast, these miRNA changes do not correlate with ACE scores and miR-34/449 does not correlate with THQ score. Notably, miRNAs-532 and 375 are two of the 9 sperm miRNAs whose enhanced levels in male mice mediate how chronic variable (CV) stress leads to a suppressed HPA axis response in offspring, a trait linked to mental health disorders. This proposal investigates how elevated levels of sperm miRNAs-532,361,375 and 491 in men with high THQ scores might affect their offspring using the mouse model that previously demonstrated how elevated levels of sperm miRNAs transmit the stress-related effects of paternal CV stress across generations. Aim 1 tests how injecting THQ-associated miRNAs, at levels found in sperm of men exposed to high levels of trauma, into mouse zygotes affects the phenotypes of resulting offspring. Aim 2 will begin to reveal how these injected miRNAs lead to phenotypic changes revealed in Aim 1 by i) identifying gene expression changes induced by them in mouse blastocysts, and ii) looking for similarities to those occurring in discarded blastocysts from IVF procedures using men’s sperm with elevated THQ associated miRNAs derived from an ongoing, independently funded project. This study has the potential to: a) provide strong support for using mouse models to understand epigenetic inheritance in humans; and b) drive future research showing that a significant portion of inherited susceptibility to mental health disorders arises from epigenetic inheritance, and how to reverse it before fatherhood.

U.S. National Science Foundation

The Tribal Colleges and Universities Program (TCUP) provides awards to Tribal Colleges and Universities, Alaska Native-serving institutions, and Native Hawaiian-serving institutions to promote high quality science, technology, engineering and mathematics (STEM) education, research, and outreach. TCUP-eligible institutions are predominantly two-year and community colleges. Support is available to TCUP-eligible institutions (see the Additional Eligibility subsection of Section IV of this solicitation) for Planning Grants, Instructional Capacity Excellence in TCUP Institutions (ICE-TI), Broadening Participation Research in STEM Education (BPR) Projects, Targeted STEM Infusion Projects (TSIP), and Catalyzing Opportunities for Research and Education (CORE). Through these mechanisms, along with collaborations with other National Science Foundation (NSF) units and its work with other organizations, TCUP aims to increase Native individuals' participation in STEM careers and the quality of STEM programs at??TCUP-eligible institutions. TCUP strongly encourages the inclusion of activities that will benefit veterans.Planning Grants provide support to undertake self-analysis of the TCUP-eligible institution's undergraduate STEM programs to identify components that need improvement or enhancement in order to ensure a high-quality??undergraduate STEM education.??Instructional Capacity Excellence in TCUP Institutions (ICE-TI) Projects (formerly Initiation Projects) provide support to design, implement??and assess comprehensive institutional efforts to increase the numbers of STEM students and the quality of their preparation by strengthening STEM education and research. ICE-TI Projects create and/or adapt and assess innovative models and materials for teaching and learning in STEM, embody knowledge about how students learn most effectively in STEM teaching and learning activities, and bring STEM disciplinary advances into the undergraduate experience. Broadening Participation Research in STEM Education (BPR) provides support for research projects that seek to create and study new models and innovations in STEM teaching and learning, enhance the understanding of the participation of diverse groups in STEM education and inform education practices and interventions.?? BPR projects add new research-based strategies and models to broadening participation in STEM and increase the capacity of scholars in TCUP-eligible institutions to conduct this type of research. Targeted STEM Infusion Projects (TSIP) provide support toward achieving a short-term, well-defined goal that promises to improve the quality of undergraduate STEM education at an eligible institution.?? Targeted STEM Infusion Projects could, for example, enhance academic infrastructure by systematically adding traditional knowledge to the scope or content of a STEM course, updating curriculum, modernizing laboratory research equipment, or improving the computational network array for research and education. Catalyzing Opportunities for Research and Education (CORE) projects provide support for??faculty members in STEM areas at??TCUP-eligible institutions to pursue research at an NSF-funded Center, at a research-intensive institution, or at a national laboratory. Awards are intended to help further the faculty member's research capability and effectiveness, to improve research and teaching at his or her home institution, and to involve undergraduate students in research experiences. These awards are particularly appropriate??as a means of recruiting and retaining??highly qualified scientists and engineers at TCUP-eligible institutions.

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

PROJECT SUMMARY Placental aging is the programmed process of progressive functional decline and tissue maturation that occurs throughout the course of pregnancy, particularly in the later stages. While this is a normal feature of development, accelerated or pathological placental aging triggered by stress can impair placental function and lead to serious complications, including stillbirth, preeclampsia, fetal growth restriction, and preterm birth. Understanding mechanisms driving premature placental aging is critical for developing strategies to improve pregnancy outcomes. This proposal tests the hypothesis that precocious trophoblast differentiation leads to exhaustion of the trophoblast stem cell (TSC) pool, triggering premature placental aging and disease. TSCs maintain placental homeostasis by regenerating the syncytiotrophoblast – the multinucleated epithelial layer that mediates nutrient and gas exchange at the maternal-fetal interface. Loss of TSC self-renewal or differentiation under stress may compromise placental regenerative capacity and contribute to aging and failure. Our preliminary data identify the transcription factor CEBPB as a key regulator of the stress response and TSC differentiation. Conversely, trophoblast-associated microRNAs, including the murine miR-290~295 cluster and its human ortholog miR- 371~373, help preserve TSC identity by supporting stem cell self-renewal and metabolism. Loss of miR-290 in mice leads to premature depletion of the TSC pool, the accumulation of aging markers in the placenta, and stillbirth – linking early stem cell dysfunction with placental failure. The overall goal of this project is to define the molecular and metabolic pathways that govern TSC differentiation and placental aging. Aim 1 will characterize stress-induced TSC differentiation and senescence, focusing on the regulatory roles of CEBPB and miR- 371~373. Aim 2 will determine how TSC depletion contributes to placental aging and stillbirth in miR-290 knockout mice, using transcriptomic and histologic approaches. Aim 3 will investigate the role of metabolic reprogramming in TSC fate and aging, and test whether metabolic interventions can preserve stemness under stress. By linking TSC biology with placental aging and adverse outcomes, this work provides a conceptual framework that opens avenues for therapeutic innovation. Applying principles from aging biology to the placenta may enable repurposing of existing anti-aging strategies to reduce placental dysfunction and prevent the most devastating consequence—stillbirth. This project directly addresses the goals of NOSI NOT-HD-23-021: The Road to Prevention of Stillbirth.

NCI - National Cancer Institute

PROJECT SUMMARY Metastasis is the leading cause of mortality among patients diagnosed with solid tumors. In this regard, identifying common mechanisms within primary, premetastatic and metastatic niches that contribute to progression of solid tumors represents a significant need. The Lipocalin-2 gene (LCN2, neutrophil gelatinase- associated lipocalin or NGAL) encodes a cytosolic and secreted protein (Lcn2) that regulates receptor trafficking, innate immune responses, inflammation, microbiome dynamics and iron homeostasis. Previous work from our group and others has shown that Lcn2 can promote stemness and tumorigenesis in models of solid tumor progression. However, uncertainty persists about the stage or stages at which LCN2 exerts tumorigenic effects and whether the mechanisms of LCN2 action vary based upon stage or tumor location. In this regard, we have recently performed unbiased single-cell spatial proteomic and transcriptomic screens of breast and pancreatic cancer patient samples leading us to define tumor cell autonomous and non-atonomous roles for Lcn2/LCN2 in promoting solid tumor progression. Based upon these findings, the central hypothesis of this proposal is that Lcn2 promotes FGFR2 signaling and adaptive anti-inflammatory immunity to support solid tumor progression. The overall objective of this proposal is to determine the spatiotemporal dynamics of Lcn2- governed tumor cell autonomous and non-autonomous mechanisms driving metastatic progression of solid tumors. Our rationale for pursing this work is that understanding these mechanisms will position Lcn2 as a biomarker for immune therapy resistance and target for improving immune therapy success in immunologically cold tumors. To test the central hypothesis, we propose to specific aims that will (I) identify the tumor cell autonomous signaling mechanisms governing Lcn2-dependent FGF2-induced tumor cell invasion and (II) define the intercellular communication mechanisms by which Lcn2 reduces inflammation and enriches regulatory T cells within the metastatic niche. The basis of this project is conceptually innovative and employs technically innovative transgenic, pharmacology, cell cycle reporter, protein reporter tagging, single-cell spatial omics, spinning disc time-lapse confocal and CRISPR methods in combination with preclinical allo- graft/xenograft models of solid tumors and hetero-multicellular cancer spheroid cultures. This work is expected to (1) provide exceptional research opportunities for undergraduates while enhancing the research environ- ment at Baylor University and (2) elucidate targetable mechanisms that govern solid tumor progres- sion/metastasis. As such, the proposed studies will have a positive impact on Baylor student success and identify novel treatment strategies to improve cancer outcomes.

NCI - National Cancer Institute

PROJECT SUMMARY Acute myeloid leukemia (AML) is an aggressive, devastating cancer with limited treatment options. AML progresses rapidly and presents significant treatment challenges due to its immunosuppressive tumor microenvironment , which impairs immune cell function. Group 1 innate lymphoid cells (ILCs), including natural killer (NK) cells and ILC1s, play key roles in immunity. ILC1s reside in tissues and were initially believed to function primarily by secreting cytokines such as IFN-γ, TNF-α, and GM-CSF. However, their anti-tumor activity has been largely unknown. In 2022, we addressed this gap and published our discovery in Nature Immunology as a cover story. We found that ILC1s isolated from AML patients are functionally impaired, whereas ILC1s from healthy mice are significantly more potent. Healthy ILC1s induce the death of leukemia stem cells (LSCs), block LSC differentiation into leukemia progenitor cells, and promote the transition of LSCs into non-leukemic lymphoid progenitors. Mechanistically, normal ILC1s target LSCs by secreting IFN-γ and engaging receptor-ligand interactions (e.g., DNAM-1–CD155 and IL-7 receptor–IL-7). Despite identifying key features of ILC1s and their role in inhibiting LSCs, important questions remain unanswered. It is still unclear how ILC1s develop in vivo under normal or AML conditions, and the mechanisms through which ILC1s induce LSC death and differentiation in humans are largely unexplored. Moreover, the therapeutic potential of ILC1s remains unknown. We hypothesize that ILC1s possess strong anti-LSC activity and unique developmental pathways, offering a novel approach to control or treat AML and potentially prevent its relapse. The goals of this project are to elucidate the mechanisms of ILC1 anti-tumor activity, characterize their developmental pathways, and explore their therapeutic applications. In Aim 1, we will dissect the mechanisms by which ILC1s induce LSC death (e.g., via pyroptosis) and drive M1 polarization of LSC-differentiated myeloid cells in humans. In Aim 2, we will characterize ILC1 developmental pathways in both normal and AML conditions. Leveraging our expertise in developing adoptive cellular therapies, including chimeric antigen receptor (CAR) NK cells for AML, in Aim 3, we will study novel FLT3-targeting CAR ILC1s that we generated. FLT3 is highly and selectively expressed on AML blasts and LSCs, making it an ideal target. We will generate allogeneic, off-the-shelf, ready-to-use FLT3- CAR ILC1s from umbilical cord blood CD34⁺ cells or by converting NK cells into ILC1s, which we demonstrated. These CAR ILC1s will be tested for their anti-AML efficacy in preclinical models and compared to unmodified ILC1s. Additionally, we will combine ILC1s or FLT3-CAR ILC1s with NK cells and an FDA-approved tyrosine kinase inhibitor, which upregulates FLT3 expression on AML cells. Finally, we propose to reprogram endogenous ILC1s by treating them with IL-7 to enhance their activity. A deeper understanding of ILC1 development and function, anticipated through the completion of this study, holds significant promise. The knowledge gained could lay the groundwork for diverse therapeutic strategies that have the potential to reduce mortality in AML patients.

U.S. Mission to Kyrgyzstan

The U.S. Department of State's Embassy Bishkek Public Diplomacy Section (PDS) announces an open competition to implement a program supporting the American Corners network in the Kyrgyz Republic the U.S. government's primary information and cultural centers. They showcase American innovation to foreign audiences and promote demand for U.S. products and services, particularly as the United States celebrates the 250th anniversary of its founding. The program will provide strategic guidance, administrative oversight, and resource support to the network of six American Corners located in Bishkek, Kant, Naryn, Karakol, Manas, and Osh serving approximately 200,000 visitors annually. Activities will enhance English language capacity through conversation clubs and educational resources; promote STEM excellence through hands-on workshops and mobile makerspaces; foster cultural exchange through speaker programs and events celebrating the 250th anniversary of U.S. independence; and maintain free, open, and reliable internet connectivity and upgraded facilities to serve as premier platforms for Embassy-driven campaigns. Target participants include emerging voices aged 14-30 (university students, young professionals, entrepreneurs, and educators), students aged 12-18 in rural areas with limited access to educational resources, and established opinion leaders and teachers who can multiply impact. Kamkor Center initiatives demonstrate American leadership in accessibility through the the Americans with Disabilities Act and promote increased access to information and public spaces by providing tailored resources and programs about the United States to the Kyrgyz public, including people with disabilities. By focusing on these priorities, this program will create long-term partnership opportunities with educational institutions and foster deeper U.S.-Kyrgyz ties in high-growth sectors. The program will increase engagement with world-leading American educational resources and technological platforms while promoting American values of free speech and innovation. By promoting increased knowledge of American society, culture, and educational and professional opportunities, the program will ensure the United States remains the partner of choice for educational exchange, cultural engagement, and technological innovation in the Kyrgyz Republic. Please follow all instructions below.

U.S. Mission to United Arab Emirates

The Public Diplomacy Section (PDS) at U.S. Mission UAE invites results-oriented proposals for programs that foster economic opportunities for U.S. businesses, investors, and innovators, and showcase American leadership and excellence in science, technology, culture, arts, sports, culinary diplomacy, artificial intelligence (AI), and health. The purpose is to strengthen ties between the United States and the United Arab Emirates in ways that make America safer, stronger, and more prosperous and advance the interests of the American people as we commemorate the 250th anniversary of U.S. independence (2026) and celebrate the enduring U.S.-UAE partnership. See Section C, Program Description for more information. This APS outlines the funding priorities and strategic themes we will focus on in FY2026, and the procedures for submitting requests for funding. Please carefully follow all instructions below. Proposals that fail to conform to the requirements outlined in this APS will not be considered. Goals and Objectives Proposals must focus on one of the seven priority areas outlined below. All proposed programs must clearly advance American strength, safety, and/or prosperity, to advance Goal 2 of the Bureau of Near Eastern Affairs: Secure opportunities to advance U.S. commercial and strategic interests. Specifically, all proposals must include an American element or demonstrate meaningful engagement with American experts, organizations, or institutions in ways that advance U.S. interests and promote understanding of American policies, perspectives, society, culture, and values. Proposals that include programming across multiple emirates are encouraged. Possible PD grant proposals include, but are not limited to: Programs that generate tangible opportunities for U.S. companies and American experts and institutions by connecting them with Emirati partners, suppliers, procurement channels, investors, government stakeholders, and/or decision makers; supporting U.S. market entry and visibility in the UAE; and facilitating engagements designed to produce measurable outcomes, including deals, partnerships, exports, investment, licensing, training agreements, or expanded U.S. market share across strategic sectors. Proposals may include activities such as events, collaborative projects, workshops, conferences, speaker series, alumni engagement initiatives, performances, trade and expo engagement, business networking activities, or programs conducted in connection with appropriate UAE trade shows, festivals, or public events. Examples of programs include, but are not limited to: Programs that connect U.S. companies and experts with Emirati partners, suppliers, procurement channels, investors, and decisionmakers, facilitating business-to-business networking, investment matchmaking, and engagements designed to produce commercial partnerships, deals, exports, or expanded market access for American firms across strategic sectors. Subject-matter expert, academic, and professional lectures, seminars, workshops, and speaker programs featuring American experts. University-industry partnerships, including with research or technology parks, that generate commercialization projects and innovation initiatives, creating opportunities for U.S. companies in STEM fields and emerging technologies to expand partnerships and commercial engagement in the UAE. Programs in partnership with UAE institutions and American companies that promote U.S. technologies, products, and services through workshops, trade show engagement, expos/showcases, and business networking in priority sectors such as AI, healthcare, energy, or the creative industries. Hackathons, innovation competitions, maker spaces, or other hands-on activities that showcase American leadership in AI, space, cybersecurity, and other emerging technologies. Professional and academic exchanges, training programs, and collaborative projects between U.S. and UAE institutions. (Note: This funding cannot be used to support construction-related activities. End note.) Priority Program Areas and Strategic Themes: Proposals must focus on one of the seven priority areas outlined below: 1. CELEBRATING AMERICA'S 250th ANNIVERSARY (FREEDOM 250): Initiatives that celebrate America s 250th birthday by showcasing American excellence, innovation, technology, dynamism, culture, and strength of the U.S.-UAE partnership. 2. CELEBRATING AMERICAN SPORTS EXCELLENCE DURING THE AMERICAN DECADE OF SPORTS: Initiatives that leverage major U.S.-hosted sporting events, such as the 2028 Summer Olympics and Paralympics and the 2034 Winter Olympics and Paralympics, to showcase American excellence in sports, sports science, athlete development, event management, tourism, and entertainment while strengthening U.S.-UAE engagement and creating opportunities for American sports institutions, companies, and experts to expand partnerships and commercial engagement in the UAE. 3. ARTIFICIAL INTELLIGENCE (AI) ADOPTION AND INNOVATION: Initiatives that promote the adoption of American AI technologies and strengthen U.S.-UAE collaboration in AI, innovation, and applied STEM fields, including cybersecurity, space, energy, water security, and smart agriculture, while creating opportunities for U.S. companies, universities, and research institutions to expand partnerships and commercial engagement in the UAE. 4. CULINARY DIPLOMACY:. Initiatives that use food and culinary arts as a bridge to strengthen U.S.-UAE ties while promoting American agricultural products, food systems, hospitality expertise, and culinary innovation. Programs should create opportunities for U.S. food producers, culinary professionals, hospitality companies, and agricultural exporters to expand partnerships and commercial engagement in the UAE. 5. CULTURAL HERITAGE PRESERVATION AND DIGITAL HERITAGE: Initiatives that showcase American leadership in cultural heritage preservation, artifact and antiquities protection, site conservation, and digital heritage technologies, including VR/AR platforms, 3D scanning, and digital archiving, while promoting U.S. expertise and expanding opportunities for American technology providers, educational institutions, and cultural organizations to strengthen partnerships and commercial engagement in the UAE. 6. HEALTH AND LIFE SCIENCES: Initiatives that strengthen U.S.-UAE collaboration in health and life sciences by promoting American expertise, technologies, research, and innovation while expanding partnerships and commercial opportunities for U.S. companies, healthcare institutions, and academic organizations in the UAE. 7. CREATIVE INDUSTRIES AND DIGITAL INNOVATION: Initiatives that strengthen U.S.-UAE cooperation and commercial engagement in the creative industries, digital media, entertainment, gaming and game development, esports, immersive technologies, and AI-enabled creative sectors by showcasing American platforms, technologies, platforms, storytelling, and innovation models while creating opportunities for U.S. companies, creators, developers, institutions, and experts to expand partnerships, collaboration, and market engagement in the UAE. Programs may also highlight the importance of intellectual property protections, creator rights, licensing frameworks, and responsible innovation in supporting growth across the digital and creative economy. In addition to the specific requirements listed above, all proposals must: Clearly indicate the primary grant priority area the program is focused on. Clearly indicate the key public diplomacy audience(s) that will be targeted by the program and the key activities to be delivered through the program. Identify the emirate(s)/city(cities) in which activities will take place. Identify specific outcomes to be achieved by the end of the grant period. Clearly delineate how elements of the proposed program will have a multiplier effect and be sustainable beyond the life of the grant. Provide a traditional and/or social media plan for marketing program activities and outcome, if applicable. Identify any tools (surveys, beneficiary interviews, focus groups, etc.) that will be developed and used for Monitoring and Evaluation purposes. Participants and Audiences: All proposals must focus on audiences in the United Arab Emirates and in the United States. If appropriate and feasible, proposals may have a regional scope and include participants from other countries, with a view to strengthening broader U.S. engagement in the region. Proposals should describe both the primary and secondary audiences for the program, including age, sex, geographic location, and anticipated reach. Primary audiences are those that will participate directly in the program, and secondary audiences include those reached indirectly, for example, via traditional or social media. Specific audiences that are considered a priority include: Students, emerging leaders, and young professionals (ages 14-45), particularly in STEM, entrepreneurship, sports, media, technology, culinary arts, and the creative industries; Entrepreneurs, innovators, researchers, educators, artists, chefs, content creators, startup founders, and other professionals engaged in technology, business, culture, and innovation sectors; Mid-career and senior-level professionals, decision-makers, institutional leaders, investors, and industry experts across sectors including artificial intelligence, healthcare, education, aerospace, sports, media, cultural heritage preservation, entertainment, and emerging technologies; Alumni of U.S. government-funded programs; and Representatives of academic institutions, research organizations, technology parks, innovation hubs, cultural institutions, and business associations involved in strengthening U.S.-UAE collaboration and commercial engagement. Mandatory application forms SF-424 (Application for Federal Assistance organizations) or SF424-I (Application for Federal Assistance --individuals) at grants.gov. SF-424A (Budget Information for Non-Construction programs) at grants.gov. SF-424B (Assurances for Non-Construction programs) at grants.gov or the Mission's website (Note: The SF-424B is only required for individuals, organizations exempt from registration, and for organizations not required to fully register in SAM.gov.) Budget Project Narrative Template: (detailed budget categories) at grants.gov. Summary Page (optional) Cover sheet stating the applicant's name and organization, proposal date, program title, program period proposed start and end date, and brief purpose of the program. Proposal (5 pages maximum) The proposal should contain sufficient information such that anyone not familiar with it would understand exactly what the applicant wants to do. You may use the Mission's recommended proposal template (APS Application Form) included with our APS package on Grants.gov or your own proposal format, but the proposal must include all the items below. Proposal Summary: Short narrative that outlines the proposed project, including project objectives and anticipated impact. Introduction to the Organization or Individual applying: A description of past and present operations, showing ability to carry out the program, including information on all previous grants from the U.S. Mission UAE and/or U.S. government agencies. Problem Statement: Clear, concise, and well-supported statement of the problem to be addressed and why the proposed program is needed Program Goals and Objectives: The "goals" describe what the program is intended to achieve. The "objectives" refer to the intermediate accomplishments on the way to the goals. These should be achievable and measurable. Project Activities: Describe the program activities and how they will help achieve the objectives. Project Methods and Design: A description of how the program is expected to work to solve the stated problem and achieve the goal. Include a logic model as appropriate. Proposed Project Schedule and Timeline: The proposed timeline for the program activities. Include the dates, times, and locations of planned activities and events. Key Personnel: Names, titles, roles and experience/qualifications of key personnel involved in the program. What proportion of their time will be used in support of this program? Project Partners: List the names and type of involvement of key partner organizations and sub-awardees. (NOTE: If your proposal requires permission, collaboration, cooperation, and/or some form of approval from or partnership with an Emirati governmental office to effectuate the project, you should obtain that permission and support prior to submitting your proposal so you can include the required letters of permission or agreement with your proposal. Project Monitoring and Evaluation Plan: Throughout the timeframe of the grant, how will the activities be monitored to ensure they are happening in a timely manner, and how will the program be evaluated to make sure it is meeting the goals of the grant? Media Outreach Strategy: Applicant's plan for using traditional and/or social media to raise awareness of the program and U.S. funding of it, recruit participants, and highlight program impact and benefits for Emiratis. Budget Justification Narrative After filling out the SF-424A Budget (above), use a separate file to describe each of the budget expenses in detail. See section I. Other Information: Guidelines for Budget Submissions below for further information. Proposal applications may be submitted at any time before the closing date of July 15, 2026, 5:00PM UAE Time at 5.00 p.m. UAE time. Applications received after the deadline will not be considered.

U.S. National Science Foundation

The NSF Research Traineeship (NRT) program seeks proposals that explore ways for graduate students in research-based master s and doctoral degree programs to develop the skills, knowledge, and competencies needed to pursue a range of STEM careers. The program is dedicated to effective training of STEM graduate students in high priority interdisciplinary or convergent research areas, through a comprehensive traineeship model that is innovative, evidence-based, and aligned with changing workforce and research needs. Proposals are requested that address any interdisciplinary or convergent research theme of national priority, as described in section II.D below. The NRT program addresses workforce development, emphasizing broad participation, and institutional capacity building needs in graduate education. The program encourages proposals that involve strategic collaborations with the private sector, non-governmental organizations (NGOs), government agencies, national laboratories, field stations, teaching and learning centers, informal science centers, and academic partners. NRT especially welcomes proposals that reflect collaborations between NRT proposals and existing NSF Eddie Bernice Johnson Inclusion across the Nation of Communities of Learners of Underrepresented Discoverers in Engineering and Science (INCLUDES) Initiative, Research Experiences for Undergraduates (REU), Louis Stokes Alliances for Minority Participation (LSAMP), NSF Scholarships in Science, Technology, Engineering, and Mathematics (S-STEM), and NSF STEM Ed Organizational Postdoctoral Fellowship program (STEM Ed OPRF) projects, provided the collaboration will strengthen both projects. Researchers at minority serving institutions and emerging research institutions are strongly encouraged to submit proposals. Collaborations between NRT proposals and existing NSF INCLUDES projects should strengthen both NRT and INCLUDES projects.

NIAMS - National Institute of Arthritis and Musculoskeletal and Skin Diseases

ABSTRACT Epidermis of the skin undergoes continuous self-renewal through a tightly regulated balance of keratinocyte proliferation and terminal differentiation. Disruption of this balance is characteristic of inflammatory skin disorders such a psoriasis, atopic dermatitis, and neutrophilic dermatoses. The etiologies of these skin disorders are complex and heterogeneous, as are the needs for treatments. Our long-term goal is to elucidate how dysregulation of K63-Ub-mediated signal transduction pathways in keratinocytes contribute to skin inflammation. Towards this end, our recent studies have focused on UBE2N, a ubiquitin conjugase that forms heterodimers with an essential noncatalytic partner, UBE2V1 or UBE2V2, to specifically catalyze K63-Ub of target proteins. We demonstrate that conditional knockout of Ube2n in mouse keratinocytes induces psoriasis- like inflammatory skin lesions with a raised and scaly appearance. Transcriptomic and histological analyses identified a diminished epidermal stem cell compartment, a thickened epidermal spinous layer, and an increased infiltration of myeloid-skewed immune cells. This is correlated with increased expressions of myeloid cell chemokines such as CXCL1 and CXCL2 and IL1 family cytokines in keratinocytes and infiltrating myeloid cells. Oral delivery of the small molecule inhibitor of IRAK1/4, common mediators of the IL1R and TLR signaling pathways, alleviated immune infiltration and epidermal defects of the mutant skin. These data highlight a key role for UBE2N in regulation of epidermal and cutaneous immune homeostasis. In line with these animal data, recent GWS studies demonstrate a causal association between UBE2V1 polymorphism and psoriasis. Together, these data support the hypothesis that UBE2N partners with UBE2V1 to restrain keratinocyte recruitment of myeloid cells through suppression of IL1 and CXCL1/2-mediated inflammatory crosstalk between keratinocytes and myeloid cells. We propose 3 specific aims to: 1) validate the importance of UBE2N catalytic function and the role of UBE2V1 in epidermal homeostasis and cutaneous immune homeostasis, 2) determine the contribution of the IL1 signaling pathway in UBE2N-null skin inflammation, and 3) assess the utility of CXCL1/2 receptor antagonists in mitigating neutrophilic dermatosis. We will utilize conditional genetic animal models along with the cutting-edge techniques of single cell transcriptomics and global proteomics to comprehensively analyze mechanistic aspects of UBE2N/UBE2V1-mediated K63-Ub in cutaneous inflammation and therapeutic targeting. Results of these studies will reveal novel mechanisms of epidermal and cutaneous immune homeostasis, as well as insights for therapeutic development.

NIDA - National Institute on Drug Abuse

PROJECT SUMMARY Cocaine use disorder (CUD) is highly comorbid in people with HIV (PWH) and can accelerate infection, alter neuropathology, and exacerbate cognitive decline despite antiretroviral therapy (ART). Many of these effects are due to the infection and dysregulation of CNS-associated myeloid cells, especially microglia, which comprise a significant reservoir in this compartment. However, the precise mechanisms by which cocaine (Coc) dysregulates microglia to enhance HIV infection are unclear, partly due to the lack of translationally relevant human microglial models suitable for mechanistic evaluation of Coc-mediated changes in viral dynamics. Classically, Coc has been thought to act by blocking dopamine transporter (DAT) activity, exposing microglia to aberrantly high dopamine concentrations. Our data show that dopamine can increase HIV infection and inflammation in microglia and other myeloid cells. However, recent data show that Coc has other mechanisms of action beyond the modulation of dopaminergic tone, involving the ER protein sigma1 (σ1), which has diverse cellular functions including the modulation of cellular stress pathways such as the unfolded protein response (UPR). Viruses, including HIV, can exploit the UPR to amplify stress-induced protein production in the host cell, enhancing viral replication. Our preliminary studies indicate that Coc’s effects on σ1 may drive a Coc-mediated increase in HIV infection in microglia, potentially through increased stress response and independent of dopamine’s effects. My preliminary data show that both Coc and σ1 agonists increase HIV replication in human inducible pluripotent stem cell (iPSC)-derived microglia (iMg). These effects are blocked by σ1 antagonism but not by inhibition of DAT or dopamine receptors. We also show increased σ1 protein expression and recruitment to the ER/nuclear envelope space in HIV-infected iMg treated with Coc, and preliminary single-cell RNAseq data suggest changes in the UPR. Therefore, we hypothesize that Coc-mediated activation of σ1 increases HIV infection of microglia via activation of the UPR. In Aim 1, we will test the involvement of σ1 in driving Coc-mediated changes in HIV infection of iMg using pharmacological and genetic modulation, and we will also confirm the absence of dopaminergic involvement. We will assess changes in viral dynamics using AlphaLISA and immunofluorescence (IF) high-content imaging. In Aim 2, we will test the hypothesis that Coc induces greater σ1 activity in the presence of HIV infection utilizing confocal and high-content IF imaging of σ1 subcellular localization in cellular compartments like the nuclear envelope, ER, and mitochondria-associated ER membrane. Movement of σ1 to these compartments is a feature of σ1 activation. In Aim 3, we will use single-cell RNAseq to test the hypothesis that Coc-induced σ1 activity drives increased HIV infection in iMg via upregulation of UPR genes. The results from these experiments will not only define novel interactions between HIV and σ1 that could reveal new antiretroviral targets but will also broadly inform on the role of σ1 in microglia and potentially identify biomarkers for prevention strategies against CUD and its associated comorbid diseases.

NHLBI - National Heart Lung and Blood Institute

PROJECT SUMMARY/ABSTRACT Mutations in telomerase cause bone marrow failure in patients suffering with dyskeratosis congenita and other associated telomere biology disorders. While mutations in these patients are found in different components of telomerase, mutations in genes that regulate the processing of the RNA component of telomerase, TERC, are the most prevalent. Due to a lack of adequate models and intrinsic difficulties in studying human telomerase in physiologically relevant cells, the molecular pathways that control TERC biogenesis and decay during hematopoiesis remain largely unknown. Progress in the field has been hampered by species and even cell-type specific differences in telomerase biology that limit our understanding of the molecular mechanisms leading to the disproportionate role of TERC in hematopoietic failure when compared to other components of telomerase. A better understanding of the molecular regulation of TERC biogenesis and function in hematopoietic cells is essential for development of novel alternatives for patients, which remain without a cure. The focus of this proposal is to use different in vitro and in vivo approaches to decipher molecular pathways controlling TERC biogenesis and decay in blood cells, as well as the function of TERC during erythroid, myeloid and lymphoid development. We have developed unique models, including targeted hematopoietic differentiation of human pluripotent stem cells, transplantation of primary CD34+ human stem cells into sub-lethally irradiated mice, and studies in primary patient samples, that will allow a complete analysis of the pathways regulating TERC decay and function during hematopoietic development. For that, two specific aims are proposed that will both identify novel regulators of TERC decay in blood cells, as well as specific functions of TERC in the hematopoietic system. Aim 1 will determine the role of novel, recently identified 3'- end RNA deadenylases to TERC processing in the blood, and to which extent different RNA deadenylases prevent TERC degradation by the exosome. We will complement these experiments with the identification of the molecular effectors of a novel route for TERC decay, triggered by differential TERC capping on its 5'- end, and mediated by trafficking to the cytoplasm. We will investigate if modulation of these different pathways can rescue hematopoietic development in telomerase mutants. Aim 2 will investigate novel functions of TERC outside telomerase that can explain the disproportionate role that mutations that affect TERC levels show in bone marrow failure. We have created unique cellular systems where we can uncouple TERC expression from telomere length, and will utilize them during hematopoietic differentiation to study direct functions of TERC on DNA damage and regulation of hematopoietic gene expression programs. These studies will determine the molecular mechanisms controlling TERC decay and function in hematopoietic cells. Our unique cellular tools, combined with our expertise in telomerase, RNA decay, and stem cell biology puts us in an ideal position to make a significant impact in this field.

NIA - National Institute on Aging

PROJECT SUMMARY This NIH F30 grant proposal investigates how somatic mutations in hematopoietic stem cells (HSCs) contribute to aging-related diseases through clonal expansion of mutated blood cells. This phenomenon, called clonal hematopoiesis, becomes more common as individuals age, affecting over 10% of in people over 60 years old. This study focuses on mosaic chromosomal alterations (mCAs), large-scale mutations that are under-studied compared to smaller mutations like clonal hematopoiesis of indeterminate potential (CHIP). mCAs are linked to lymphoid malignancies and infection susceptibility due to lymphoid-biased differentiation, while CHIP correlates with cardiovascular diseases and myeloid malignancies through myeloid-biased differentiation. I hypothesize that mCA expansion is determined by individual factors rather than mCA genetic change and that clones with greater expansion rates will have increased disease risk. Aim 1 examines the influence of mCA characteristics and environmental factors on clonal expansion rates using longitudinal blood samples from 30,000 individuals in Vanderbilt’s BioVU genomic and clinical biobank. Using longitudinal mCA trajectories, I will quantify the contribution of the mCA mutation and individual characteristics (e.g., age, sex, BMI, smoking, type 2 diabetes, lipoprotein levels) to clonal expansion rate and build a predictive model for mCA clonal expansion. My working hypothesis for Aim 1 is that mCA expansion varies widely among individuals with the same mCA and thus modifiable lifestyle exposures are major contributors to clonal expansion rate. The longitudinal samples in BioVU will not be sufficient to test genetic and phenotypic associations with clonal expansion rate. Therefore, Aim 2 expands the study to detect mCAs in > 1 million individuals across various genomic biobanks with single blood draws (i.e., NHLBI TOPMed, NIH All of Us, UK Biobank, and BioVU). To determine mCA clonal expansion rate from a single timepoint, I will apply Passenger-Approximated Clonal Expansion Rate (PACER), which estimates mCA expansion rate from a single blood draw to build upon my measured mCA analysis by two orders of magnitude. A genome-wide association study and a phenome-wide association study will be conducted to identify germline variants and phenotypic correlations related to mCA clonal expansion rates. My working hypothesis for Aim 2 is that 1) certain germline variants predispose individuals to faster mCA growth and 2) specific disease phenotypes, including chronic lymphocytic leukemia and infection susceptibility, are associated with faster mCA clonal expansion rate. This research aims to significantly enhance our understanding of mCA clonal expansion, addressing a fundamental biological mechanism of aging to prevent multiple diseases. Collectively, these insights will contribute to mCA risk prediction models and highlight potential biological pathways or lifestyle strategies to slow mCA expansion.