Search Grants — Free, No Account Required

NINDS - National Institute of Neurological Disorders and Stroke

PROJECT SUMMARY CLN3 disease is a rare lysosomal disease affecting approximately 1 in 100,000 live born children. It is caused by recessive mutations in the CLN3 gene, which encodes a transmembrane protein that primarily localizes to the lysosome. Affected children suffer from progressive blindness, seizures, psychosis, and cognitive and motor failure, and the disease is invariably fatal. CLN3 is implicated in various cellular processes including endolysosomal trafficking and lipid metabolism, but the primary function remains incompletely resolved. Interestingly, immune system changes have been described in CLN3 patients and animal models including early neuroinflammation in brain regions that later see the first neuronal cell dropout, suggesting the neuroimmune system plays a role in the neurodegenerative disease process. In a proteomics study of CLN3- deficient microglia, we recently discovered a dramatic elevation in the levels of the Tweety homolog protein, TTYH3, which was over 10-fold elevated in microglia isolated from presymptomatic mice, and over 20-fold elevated in microglia from symptomatic mice, suggesting TTYH3 elevation is a relatively early disease event and that it progresses with disease severity. Indeed, we also identified TTYH3 elevation in a neuronal progenitor cell model of CLN3 disease, indicating TTYH3 levels increase in response to loss of CLN3 function in both neurons and microglia. In this proposal, which is responsive to the NOFO for research projects of understudied proteins linked to rare disease (NOFO PAR-25-122), we aim to develop important tools to study the TTYH3 protein in the context of CLN3 disease. We hypothesize that TTYH3 is a novel lysosomal lipid transporter, and we will test this hypothesis by studying TTYH3 subcellular localization and by establishing Ttyh3/TTYH3 knockout mouse and human induced pluripotent stem (iPS) cell-based models that will be phenotyped to evaluate lysosomal function. Finally, we will evaluate whether modulation of TTYH3 impacts CLN3 disease pathophysiology, setting the stage for future work to fully uncover TTYH3 function and whether targeting TTYH3 in CLN3 disease holds therapeutic promise.

NCI - National Cancer Institute

PROJECT SUMMARY Acute myeloid leukemia (AML) is a genetically and cellularly heterogenous disease characterized by the expansion of hematopoietic cells across a range of cell states from stem-like cells to differentiated myeloid cells. The most mutated genes in AML are DNMT3A, NPM1 and the receptor tyrosine kinase FLT3. Despite early clinical responses, most patients relapse, and FLT3-mutant clones are not always eradicated. Our lab has developed genetically engineered mouse models of acute myeloid leukemia that are capable of activating mutations in Flt3 with Dre-recombinase, and then genetically reverting them with Cre-recombinase. We have used these models to benchmark Flt3 oncogene-addiction against best-in-class small molecule kinase inhibitors of FLT3, observing difference in disease remission and relapse. These studies have refined our interest on identifying which cells along the hematopoietic hierarchy are capable of driving relapse and which molecular pathways underlie their survival following chemical/genetic inhibition of FLT3. The major goal of this proposal is to understand the cellular mechanisms that maintain FLT3-mutant clone persistence during targeted therapy. Our preliminary data indicate that Flt3-inhibtion results in a profound differentiation response and induction of Type I Interferon signaling. We will complete integrative studies with human specimen and our innovative multi- recombinase mouse models of leukemia to derive clinically meaningful insights from mechanistic observations in model systems. In aim 1 we will determine which cells are capable of propagating leukemic disease and resolve cellular reservoirs of leukemic stem cell activity. We will perform these studies using genetically engineered mouse models, serial transplantation of purified cell populations, and functional cell ablation studies. We hypothesize that FLT3-inhibtion induced differentiation generates mature cells that are capable of reacquiring stem-like properties and drive relapse. These studies will resolve which cells are necessary to eliminate to prevent leukemic recurrence and provide a focusing lens for improving targeted therapy and relapse detection. In aim 2 we will determine the role of Type I Interferon signaling in differentiation and relapse using gain/loss of function systems. We will evaluate the therapeutic potential of interferon treatment in conjunction with FLT3 kinase inhibition. Finally, we will assess the clonal diversity of leukemic cell states using lentiviral barcoding and single cell RNA sequencing to evaluate which cells can induce an Interferon response, and what their long-term fate is following treatment. We hypothesize that Interferon signaling is necessary to potentiate FLT3-inhibitor driven differentiation, and that combined treatment will extend survival. We anticipate that these studies will more broadly inform the intersection between inflammation and differentiation in AML therapy.

NIEHS - National Institute of Environmental Health Sciences

Project Summary Autism Spectrum Disorder (ASD) is a complex neurodevelopmental condition affecting 1 in 36 U.S. children annually. While genetic, epigenetic, and environmental factors contribute to ASD, the mechanisms by which environmental exposures disrupt neurodevelopment remain poorly understood. Emerging evidence highlights the role of epitranscriptomic modifications, such as N6-methyladenosine (m6A), in regulating brain development and synaptic plasticity. This study investigates how prenatal environmental exposures (e.g., cadmium, PFAS) and protective factors (e.g., folic acid) disrupt epitranscriptomic crosstalk, contributing to ASD pathogenesis. Using the MARBLES and EARLI cohorts, we will analyze maternal exposure data and biospecimens to identify exposure-specific epitranscriptomic signatures. Brain organoids derived from induced pluripotent stem cells (iPSCs) will model the effects of environmental toxicants and folic acid on neurodevelopment and m6A regulation. Multi-omics approaches, including RNA-Seq, DNA methylation assays, and MeRIP-Seq, will uncover molecular changes linked to ASD. Machine learning algorithms will integrate multi-omics data to develop predictive models for ASD risk and severity. We propose to investigate four aims: Investigate prenatal environmental exposures’ effects on RNA modifications and DNA methylation; Assess sex-specific epitranscriptomic modifications linked to ASD phenotypes; Evaluate exposure impacts on ASD-associated epitranscriptomic changes using brain organoids; Develop predictive models for ASD risk based on disrupted epitranscriptomic crosstalks. This study bridges human epidemiologic data with organoid modeling to explore the etiology of ASD. By focusing on environmental exposures and protective factors, it addresses critical knowledge gaps and provides a foundation for non-invasive screening tools and targeted interventions. The findings will advance understanding of how environmental factors interact with epitranscriptomic regulation to influence neurodevelopment, paving the way for precision medicine strategies to mitigate ASD risk.

NCI - National Cancer Institute

SUMMARY ABSTRACT: Cancer stem cells (CSCs) are a small population of cancer cells that are typically quiescent, but capable of self- renewal and tumor initiation. Non-CSCs, which make up the majority of the cancer cell population, are cells that are constantly dividing. Since CSCs are non-dividing, they are resistant to standard chemotherapy and radiation therapy, which only target actively dividing cells. Persistence of CSCs after therapy can thus result in disease relapse. Importantly, our recent work showed that programs of stemness are activated in tumor cells as they disseminate from primary tumors. Therefore, understanding what influences the formation of disseminating CSCs may provide new potential targets for therapeutic interventions of metastasis. Metastasis is a multi-step process and, in each step, disseminating tumor cells (DTCs) encounter different physical constrictions (e.g., physical confinement, solid stress, matrix stiffness, interstitial fluid pressure, and shear stress) that may affect their behavior. Recent reports in the literature indicate that one pathway for induction of programs of stemness is through the YAP/TAZ mechanotransduction signaling pathway, indicating that physical forces may, in part, be responsible for the induction of stemness in DTCs. This project will investigate the role physical constriction plays in stem cell induction by modelling the different physical constrictions that DTCs experience as they disseminate from primary tumors. This project will draw upon engineering and biological sciences to combine a unique, validated, fluorescent biosensor for stemness with advanced microfabricated microfluidic in vitro assays, and state-of-the-art intravital imaging of the live murine lung. We propose to use this combination to study the influence of physical forces on the selection, induction, and/or sustainment of metastasizing cancer stem cells, and evaluate their retention, survival, and extravasation efficiency in the in vivo lung.

NCI - National Cancer Institute

Summary Melanoma cells exhibit remarkable plasticity, transitioning between melanocytic/proliferative and mesenchymal/invasive states. This plasticity drives intra-tumoral heterogeneity, disease progression, and therapeutic resistance. While the Microphthalmia-associated Transcription Factor (MITF) is a well-established driver of the melanocytic/proliferative phenotype, the mesenchymal/invasive phenotype and the mechanisms governing state transitions remain poorly understood. Our published and preliminary data identify the MITF paralog TFE3 and the pioneer factor TFAP2B as critical regulators of the mesenchymal state in melanoma. We identify two TFE3 isoforms, a short length (TFE3sl) and a full-length (TFE3fl), with TFE3fl specifically stabilized in invasive melanoma. Pan-TFE3 knockout reduced invasion and distal lung seeding of MITF-low melanoma cell lines. Moreover, MITF directly suppresses the mesenchymal phenotype by activating FNIP2, a key component of the non-canonical mTORC1 signaling pathway. This pathway facilitates lysosomal degradation of TFE3fl, leading us to hypothesize that TFE3fl drives metastasis. These findings suggest a molecular mechanism in which the balance of transcriptional regulation by MITF and TFE3fl, in combination with TFAP2B, governs melanoma plasticity. We hypothesize that melanoma cell plasticity is regulated by a TFE3fl-TFAP2B axis that promotes mesenchymal phenotypes and metastatic progression. To test this hypothesis, we propose the following aims: (1) identify the TFE3 isoform critical for melanoma plasticity and progression and (2) determine how the TFAP2B/TFE3fl axis establishes mesenchymal states within primary tumors. To achieve these aims, we will use patient-derived melanoma cell lines and complementary in vivo models. Zebrafish are uniquely suited for this work because we have generated a primary melanoma model that enables direct visualization of vertical tumor cell invasion in real time, combined with histological validation across multiple animals, providing high-throughput assessment of invasive behavior in the correct anatomical context. In parallel, mouse models will be essential for assessing cancer stem cell (CSC) dynamics, including tumor cell survival and dissemination within the bloodstream, as well as long-term metastatic colonization and survival outcomes. Together, these approaches will allow us to define how TFE3 and TFAP2B regulate melanoma plasticity in both early invasive transitions and later metastatic progression. This study will advance our understanding of melanoma plasticity and identify mechanisms that facilitate the adaptive nature of melanoma cells. Use of mice and zebrafish in this proposal: The rationale for utilizing mouse and zebrafish models is that normal and pathological cells behave differently in vivo due to a complex microenvironment that cannot be replicated in other model systems. Zebrafish have the added benefit of real time imaging of melanoma onset and progression

NEI - National Eye Institute

Abstract This proposal addresses the lack of human retina cell models for mechanistic and therapeutic studies. Retinal diseases are a significant cause of vision impairment in humans and account for > 54% of blindness in the USA. Despite this, there are limited or no treatment options available for several retinal diseases. Most studies on human retinal diseases have so far used animal models, including the commonly used transgenic mice that are genetically engineered to carry the disease- related gene. Although these mouse models have provided important insights into the plausible disease mechanisms of vision loss in specific retinal diseases, due to the lack of physiological human retina cell models, there has been limited translatability of molecular and therapeutic discoveries identified in the non-primate models of human eye diseases. Furthermore, from the perspective of cell-based therapies of retinal degenerative diseases that predominantly affect the photoreceptor-retinal pigment epithelium (RPE) complex in the retina, a “planar tissue comprising the photoreceptor-RPE layers” that can integrate into the existing retina is an urgent unmet translational need. To complement in vivo animal model studies and develop human retina tissue for molecular and therapeutic applications, in this project, we will utilize a combination of human induced pluripotent stem cells (hiPSCs), engineered extracellular matrix (eECM), microbubble arrays, and microfluidics technology. Aim 1 experiments will develop a microphysiological model of comprehensive hiPSC- retina that will emulate the physiological spherical geometry of the human retina in vivo and thus will be suitable for disease modeling, molecular, and drug testing applications. Aim 2 experiments will develop a planar hiPSC-photoreceptor-RPE tissue for use in cell-based therapy targeting photoreceptor and RPE loss in retinal degenerative diseases. Ultimately, hiPSC- derived comprehensive retina (Aim 1) and planar photoreceptor-RPE (Aim 2) will be a tissue engineering breakthrough for in vitro (e.g., molecular, therapeutic testing) and in vivo (cell-based therapy) applications targeting human retinal diseases.

NCATS - National Center for Advancing Translational Sciences

Abstract Support is requested for a Keystone Symposia conference entitled “iPSCs: Progress, Opportunities, and Challenges,” organized by Drs. Shinya Yamanaka, Yanhong Shi and Yasushi Kajii, with scientific programming input from Keystone Symposia. The meeting will take place January 26–29, 2026 at the International Conference Center (ICC) Kyoto in Kyoto, Japan. This conference is being held to mark the 20th anniversary of breakthrough discoveries in induced pluripotent stem cell (iPSC) technologies, which have matured into viable platforms for embryology and disease modeling, drug discovery, and cell-based therapy development for a variety of human diseases. Moreover, the combination of iPSC technology with three-dimensional organoids, organ-on-chip and the emerging technologies of AI and machine learning ensures that iPSC-based platforms will yield new applications in biomedical and translational science. These innovative technologies and their applications are the primary focus of this meeting. Therefore, this conference has been designed to gather the leading scientists from academia and industry to push forward basic knowledge and medical application of iPSCs, especially those being tested in clinical trials worldwide, toward new insights and potential drugs. Additionally, this Keystone Symposia conference will provide a unique opportunity for researchers, clinicians, industrial experts and investors to interact, creating unusual collaborative prospects. Moreover, this conference will provide a rare opportunity for attendees to hear from renowned stem cell researcher and Nobel Laureate, Dr. Shinya Yamanaka, who will be giving the Keynote Address and is one of the meeting organizers. The sharing of knowledge at this meeting is expected to be transformative for the field and lead to the development of new cellular platforms and therapeutic products, which will ultimately impact clinical practice favorably.

NHLBI - National Heart Lung and Blood Institute

The ISEH Annual Scientific Meeting serves as a premier international forum for presenting and discussing the latest research in experimental hematology, including the understanding of normal hematopoiesis, non-malignant hematopoietic diseases, as well as developmental and HSC biology. For example, topics discussed at the ISEH meeting include sickle cell disease, hematopoiesis, bone marrow niche, cell cycle, apoptosis, and signaling. The discoveries reported at the ISEH Annual Scientific Meeting are fundamental to understanding and treating blood disorders, including but not limited to anemias, bone marrow failure syndromes, and leukemia. These topics align closely with NHLBI’s mission to advance scientific understanding and develop therapies for blood diseases, including stem cell biology and gene/cell-based treatments to repair and regenerate tissues. The ISEH Annual Scientific Meeting is intentionally designed to foster scientific exchange through a variety of presentation formats, including invited lectures, short talks selected from abstracts, featured poster teasers, and traditional poster sessions. Beyond showcasing the latest research findings, the meeting cultivates an environment where researchers are encouraged to ask challenging questions and build future collaborations that drive scientific discovery forward. ISEH places a strong emphasis on fostering early-career scientists (students, postdoctoral fellows, and junior faculty), as evidenced in meeting attendance. Between 2022 and 2025, PhD students, postdoctoral fellows, and junior faculty made up an average of 61% of attendees at the ISEH Annual Scientific Meeting. Of this young investigator attendee population, an average of 48% of these PhD students, postdoctoral fellows, and junior faculty attending the ISEH Annual Scientific Meeting were based in the United States, demonstrating strong participation of US-based junior investigators. Dedicated programming includes pre-meeting workshops, career development sessions, junior faculty career forums, innovating technology presentations, networking mixers, and featured poster presentations. Additionally, our travel grant program helps defray costs for young investigators presenting abstracts, ensuring broad participation in scientific exchange. In 2026, we are adding a third track of concurrent presentations to provide additional opportunities for trainees and junior faculty to present their work. This R13 grant application requests funds in support of trainees and early career investigators as well as accessibility focused activities at the 2026 scientific meeting totaling $41,064. This includes support for travel grants, awards, closed captioning services, as well as supplies needed for trainee/early-career investigator focused activities such as abstract submission, and poster board rental.

NIDDK - National Institute of Diabetes and Digestive and Kidney Diseases

PROJECT SUMMARY/ABSTRACT Because it is unsafe to access human pancreatic islets from living donors, surrogate experimental systems are needed to answer important questions about the mechanisms through which insulin-producing β cells are destroyed in individuals who develop type 1 diabetes (T1D). Protocols for differentiating induced pluripotent stem cells (iPSCs) into islet-like clusters (SC-islets) provide a replenishable source of beta cells and are a promising alternative means for modelling interactions between human islet endocrine cells and immune cells. However, currently available biomimetic systems are not able to maintain the long-term viability of SC-islets and are not isogenic and therefore, unable to accurately model autoimmune interactions. To meet this need, this project will develop a vascularized 3D biomimetic microphysiological system (MPS) that will allow fully isogenic modelling of interactions between islets and immune cells in prolonged culture. Our basis for this model system is a proven perfusion-capable microfluidic skin-on-chip platform. This plexiglass-based chamber system has an open well on the top, which is readily adaptable to create an ideal system for culturing SC-islets. A microchannel network within the chamber promotes the formation of a vascular network in a supporting matrix. The system has been designed with inlet and outlet ports for perfusing endothelial cells, medium, cytokines, or immune cells. Furthermore, the system is configured to allow live imaging and removal of SC-islets and immune cells from the system for downstream analysis. We predict that this approach will overcome some of the described limitations of existing SC-islet culture systems and will allow mechanistic interrogation of mechanisms that promote sustained autoimmunity and pathologic interactions between SC-islets and autoreactive T cells. We will fully implement this system and demonstrate its suitability for studying interactions between human SC-islets and autoreactive T lymphocytes and then utilize it to ask specific questions about the effects of inflammatory stress on SC-islet phenotype. Importantly, our experiments will utilize T cell lines and T cell receptor sequences obtained from pancreatic organ donors with T1D, as these represent the most relevant T cells for mechanistic studies. Specifically, we will investigate the effects of inflammatory stress on islet phenotype, function and interactions with autoreactive T cells, first using 3D cultures (suitable for modeling short-term inflammatory stress) and then in the islet-on-chip system (suitable for short and long-term inflammatory stress). This will enable us to test the hypothesis that inflammatory stress alters beta cell phenotype and drives increased immune perception during the development of T1D. In addition, we will utilize the islet-on-chip system to investigate the role that the membrane repair pathway plays in dictating the vulnerability of beta cells to immune attack. We anticipate that modeling interactions between SC-islets and cytotoxic T cells will reveal a crucial role of the membrane repair pathway in determining vulnerability to immune attack. These insights are likely to suggest novel pathways that can be leveraged to treat T1D.

U.S. National Science Foundation

Revolutionizing Engineering Departments (hereinafter referred to as RED) is designed to build upon previous efforts in engineering education research. Specifically, previous and ongoing evaluations of the NSF Engineering Education and Centers Division program and its predecessors, as well as those related programs in the Directorate for STEM Education, have shown that prior investments have significantly improved the first year of engineering students experiences, incorporating engineering material, active learning approaches, design instruction, and a broad introduction to professional skills and a sense of professional practice giving students an idea of what it means to become an engineer. Similarly, the senior year has seen notable change through capstone design experiences, which ask students to synthesize the technical knowledge, skills, and abilities they have gained with professional capacities, using reflective judgment to make decisions and communicate these effectively. However, this ideal of the senior year has not yet been fully realized, because many of the competencies required in capstone design, or required of professional engineers, are only partially introduced in the first year and not carried forward with significant emphasis through the sophomore and junior years. The Directorates for Engineering (ENG) and STEM Education (EDU) are funding projects as part of the RED program, in alignment with the Improving Undergraduate STEM Education (IUSE) framework and Professional Formation of Engineers (PFE) initiative. These projects are designing revolutionary new approaches to engineering education, ranging from changing the canon of engineering to fundamentally altering the way courses are structured to creating new departmental structures and educational collaborations with industry. A common thread across these projects is a focus on organizational and cultural change within the departments, involving students, faculty, staff, and industry in rethinking what it means to provide an engineering program. In order to continue to catalyze revolutionary approaches, while expanding the reach of those that have proved efficacious in particular contexts, the RED program supports four tracks: RED Planning (Track 1), RED Adaptation and Implementation (Track 2), RED Innovation (Track 3), and RED Innovation Partnerships (Track 4). Two- and four-year institutions are encouraged to submit to any track as appropriate for their goals and context. RED Planning (Track 1) projects will support capacity-building activities at institutions of special interest to NSF s mission, including two-year engineering-centered programs building transfer partnerships, two-year or four-year institutions in EPSCoR jurisdictions, Primarily Undergraduate Institutions (PUIs), and Institutions of Higher Education (IHEs)in order to create more opportunities in engineering for students in every part of the country. Planning projects should provide the support for such institutions to explore the development of a RED Projects in Tracks 2, 3, & 4. RED Adaptation and Implementation (Track 2) projects will adapt and implement evidence-based organizational change strategies and actions to the local context, which helps propagate this transformation of undergraduate engineering education. RED Innovation (Track 3) projects will develop new, revolutionary approaches and change strategies that enable the transformation of undergraduate engineering education. RED Innovation Partnerships (Track 4) projects will achieve the same goals as Track 3 projects across multiple institutions. Of particular interest to this track are projects partnering two-year institutions with other eligible institutions. Projects in tracks 2, 3, & 4 will include consideration of the cultural, organizational, structural, and pedagogical changes needed to transform one or more departments to ones in which students are engaged, develop their technical and professional skills, and establish identities as professional engineers or technologists. The focus of projects in these tracks should be on the department s disciplinary courses and program. RED project initiatives are expected to be institutionalized at the end of the funding period. Proposals are especially encouraged that address areas of increased national interest including but not limited to advanced manufacturing, advanced wireless, artificial intelligence, biotechnology, microelectronics and semiconductors, net zero technologies, sustainability, systems engineering, and quantum engineering.

U.S. Mission to Myanmar

1. Project Background, Goals and Objectives Jefferson Center Mandalay Small Grants projects must clearly advance America First foreign policy principles by demonstrating how the proposed activities make the United States safer, stronger, and more prosperous, while celebrating Freedom250 and American excellence. Priority will be given to projects that deliver tangible, measurable benefits to U.S. interests; elevate U.S. leadership and credibility; promote reciprocal and mutually beneficial engagement with Burma and focus on one of the priority areas outlined below. AMERICA FIRST: MAKING AMERICA SAFER Programs that directly advance the security and resilience of the United States by promoting American democratic governance and interests in Burma. This includes initiatives that reinforce the rule of law, counter trafficking, digital freedom, anti-scam awareness and corruption that threaten U.S. interests, uphold American-defined human rights, and empower civil society to create an environment that aligns with U.S. peace and security priorities in the region. AMERICA FIRST: MAKING AMERICA STRONGER Programs that advance America First priorities by equipping Burma s students and young adults with skill based and vocational training that strengthens U.S. relevant economic competitiveness. Proposals should promote accurate understanding of U.S. education, institutions, and culture; prepare potential qualified candidates for lawful study and exchange opportunities related to the United States; and build durable linkages with American academic, vocational, and cultural institutions. These may also include activities that strengthen the United States global leadership by promoting American values, and civic engagement rooted in U.S. principles. These programs should deepen U.S. influence in Burma, reinforce American cultural and educational standards, and build enduring partnerships that serve American diplomatic and strategic interests. AMERICA FIRST: MAKING AMERICA MORE PROSPEROUS Projects that advance U.S. Burma economic ties and U.S. prosperity by strengthening entrepreneurs and businesses that align with U.S. commercial and strategic interests. Activities that expand economic opportunities for the United States by fostering innovation, entrepreneurship, and workforce development that benefit American businesses and industries. Priority will be given to programs that promote U.S.-led STEM education, vocational training aligned with American economic priorities, or trade capacity building that directly supports U.S. economic growth and reduces reliance on adversarial actors. AMERICA FIRST: AMERICAN EXCELLENCE Projects that showcase the superiority of American leadership, innovation, arts, and community service. These initiatives should highlight U.S. achievements and role models, inspire admiration for American values, and promote collaboration that advances U.S. interests in technology, entrepreneurship, education, and the arts, ensuring America s continued prominence on the global stage. Activities may include programs that feature U.S. experts or content on Indo Pacific strategy, maritime and economic security, or resilient supply chains, and that clearly communicate U.S. strengths, values, and strategic objectives to Burma based audiences. ENGLISH LANGUAGE LEARNING Programs that advance America First priorities by providing Business English and English for Entrepreneurship essential to U.S. linked trade, investment, and regional stability. Proposals should build high level English skills needed to work with American companies, navigate U.S. standards and contracts, participate in regional supply chains, and engage in lawful, rules based economic activity. Activities may include targeted English training for professionals, entrepreneurs, and future business leaders that uses U.S. materials, terminology, and case studies and clearly supports U.S. economic and strategic interests in the Indo Pacific. Project Audiences: Youth and Young Adults: Including students, recent graduates, and emerging professionals in both urban centers and rural regions of Burma, with a focus on those from underserved or marginalized communities in Mandalay. Civil Society Organizations: Local NGOs, advocacy groups, and community-based organizations in Mandalay, engaged in promoting democratic governance, human rights, anti-corruption, and rule of law reforms. Commercial Advancement: Individuals and entities involved in innovation, trade, and workforce development that align with U.S.-Burma economic ties and regional supply chains. Educational and Cultural Institutions: Schools, vocational training centers, universities, and cultural organizations that facilitate bilateral exchanges and promote American cultural and educational standards. Project Goals: Advance democratic governance and the rule of law in Burma by empowering civil society organizations to promote human rights, anti-corruption, and digital freedoms aligned with U.S. peace and security priorities. Enhance the skills and employability of Burmese youth and young adults through vocational training and English language programs that prepare them for lawful educational exchanges and economic participation linked to the United States. Strengthen U.S.-Burma economic ties by supporting entrepreneurship, innovation, and workforce development programs that foster trade capacity building and reduce reliance on adversarial actors. Promote American cultural excellence and leadership by facilitating educational, artistic, and community service initiatives that highlight U.S. values and strategic objectives in the Indo-Pacific region. Support sustainable, measurable outcomes by encouraging projects that incorporate robust monitoring, evaluation, and reporting mechanisms to demonstrate progress toward U.S. foreign policy goals. Project Objectives: Objective 1: Promote American Democratic Governance Enhance the institutional and operational capacity of at least 10 civil society organizations over a 12-month period to effectively advocate for rule of law, anti-trafficking measures, digital freedoms, and anti-corruption reforms. Success will be measured by the adoption and implementation of key organizational policies (e.g., financial management, transparency protocols), increased advocacy activities, and demonstrable influence on public policy aligned with U.S. peace and security priorities. Objective 2: Empower Youth through Vocational and English Language Training Provide vocational skills development and high-level English language instruction to a minimum of 1,500 youth and young adults, improving their readiness for lawful educational exchanges, workforce participation, and engagement with U.S.-linked economic opportunities. Progress will be assessed via standardized skill assessments, participant retention rates, and post-training employment or education placement data. Objective 3: Commercial Advancement and Economic Competitiveness Support at least 200 entrepreneurs and small businesses through training, mentorship, and capacity-building activities that promote innovation, STEM education, and trade capacity building. Programs should align with American economic priorities by enhancing workforce skills, facilitating access to U.S. markets, and reducing reliance on adversarial actors. Indicators of success include business growth metrics, increased participation in regional supply chains, and measurable expansion of U.S.-Burma commercial ties. Objective 4: Showcase American Excellence and Values Develop and implement cultural, educational, and leadership programs that highlight American innovation, arts, community service, and strategic expertise in areas such as Indo-Pacific security, maritime resilience, and economic policy. These initiatives should engage local audiences, promote admiration for American values, and strengthen bilateral cultural ties. Success will be measured by audience reach, participant feedback, and enhanced understanding of U.S. strategic objectives. Note: Please see detail information by clicking Related Document tab.

NIAMS - National Institute of Arthritis and Musculoskeletal and Skin Diseases

Project Summary/Abstract Muscle stem cells (MuSCs) provide myofibers with a robust tool for regeneration after injury. The efficiency of this muscle regenerating capacity depends upon the ability of MuSCs to integrate signals emanating from complementary cell types within the damaged muscle. Unfortunately, integration of these signals is often disrupted in disease, which results in a functional depletion of the MuSC population. MuSCs are still present, but they are not able to respond to regenerative cues from the environment. Although substantial evidence supports a role for epigenetic enzymes in orchestrating the transcriptional programs necessary for MuSCs to respond to niche-derived cues, how these enzymes enable MuSCs to actively shape the regenerative environment— particularly under inflammatory conditions—remains poorly understood. Our preliminary studies indicate that the H3K27 demethylase JMJD3 is rapidly induced in MuSCs after injury and regulates key genes that facilitate communication with immune cells, allowing MuSCs to exit quiescence and support tissue repair. Loss of JMJD3 in MuSCs leads to aberrant cytokine expression and excessive accumulation of inflammatory macrophages, suggesting that JMJD3 enables MuSCs not only to respond to signals but also to broadcast critical cues that help calibrate the immune response. Despite this emerging evidence, the underlying mechanisms through which JMJD3 governs MuSC–immune cell communication remain unknown. The overall objective of this project is to define the JMJD3-dependent transcriptional and epigenetic programs that allow MuSCs to modulate the inflammatory niche and initiate regeneration. We hypothesize that JMJD3 integrates signals from the regenerative environment by removing repressive H3K27me3 marks at immunomodulatory genes, permitting their expression to shape immune cell behavior and ensure efficient repair. We will address this through two aims: Aim 1: Use TEA-seq, a trimodal single-cell approach, to uncover the signaling pathways by which MuSCs regulate the magnitude and duration of the immune response to muscle injury. Aim 2: Determine how JMJD3-mediated H3K27 demethylation integrates niche-derived signals to control inflammatory resolution, testing whether modulation of H3K27me3 levels governs expression of MuSC immunomodulatory genes. Successful completion of these studies will elucidate how JMJD3 enables MuSCs to coordinate the inflammatory landscape necessary for regeneration, fundamentally advancing our understanding of how stem cells navigate and shape complex tissue environments. By revealing the extent to which epigenetic control of MuSC–immune communication influences regeneration, this work will provide a critical foundation for future efforts to fine-tune inflammation in muscle-wasting diseases.

Deere Foundation

Invests in workforce development, STEM education, and community prosperity in agricultural and rural regions.

U.S. National Science Foundation

The Launching of Early-Career Academic Pathways in the Mathematical and Physical Sciences (LEAPS-MPS) supports the launch of the careers of pre-tenure faculty whose research is in Mathematical and Physical Sciences (MPS) fields at institutions that do not traditionally receive significant amounts of MPS funding, such as Carnegie Research 2 (R2) universities, minority-serving institutions (MSIs), predominantly undergraduate institutions (PUIs). The LEAPS awards enable PIs from these institutions to initiate productive research programs and generate results useful for preparing subsequent competitive proposals to traditional NSF funding opportunities, such as a core program or a CAREER solicitation. A critical goal of the LEAPS-MPS Program is to develop the 21st-century STEM workforce representative of society s full spectrum of talent by increasing the participation in STEM research of members of communities underrepresented and/or underserved in STEM and the number of members of these communities who can serve as role models. Awards are for 24 months with budgets of up to $250,000 total costs (direct plus indirect). Proposals in response to this solicitation must be submitted for consideration tothe appropriate program in one of the five MPS Divisions.

U.S. National Science Foundation

The Law &amp; Science Program considers proposals that address social scientific studies of law and law-like systems of rules, as wellas studies of how science and technology are applied in legal contexts.The Program is inherently interdisciplinary and multi-methodological.Successful proposals describe research that advances scientific theory and understanding of the connections between human behavior and law, legal institutions, or legal processes; or the interactions of law and basic sciences, including biology, computer and information sciences, STEM education, engineering, geosciences, and math and physical sciences.Scientific studies of law often approach law as dynamic, interacting with multiple arenas, and with the participation of multiple actors.Fields of study include many disciplines, and often address problems including, though not limited, to: <ul type="disc"> <li>Crime, Violence, and Policing</li> <li>Cyberspace</li> <li>Economic Issues</li> <li>Environmental Science</li> <li>Evidentiary Issues</li> <li>Forensic Science</li> <li>Governance and Courts</li> <li>Human Rights and Comparative Law</li> <li>Information Technology</li> <li>Legal and Ethical Issues related to Science</li> <li>Legal Decision Making</li> <li>Legal Mobilization and Conceptions of Justice</li> <li>Litigation and the Legal Profession</li> <li>Punishment and Corrections</li> <li>Regulation and Facilitation of Biotechnology (e.g., Gene Editing, Gene Testing, Synthetic Biology) and Other Emerging Sciences and Technologies</li> <li>Use of Science in the Legal Processes</li> </ul> LS supports the following types of proposals: <ul type="disc"> <li>Standard Research Grants and Grants for Collaborative Research</li> <li>Conference Awards</li> </ul> LS also participates in a number of specialized funding opportunities through NSF&rsquo;s cross-cutting and cross-directorate activities, including, for example: <ul type="disc"> <li>Faculty Early Career Development (CAREER) Program</li> <li>Research Experiences for Undergraduates (REU)</li> <li>Research at Undergraduate Institutions (RUI)</li> <li>Grants for Rapid Response Research (RAPID)</li> <li>Early-concept Grants for Exploratory Research (EAGER)</li> </ul> For information about these and other programs, please visit the <a href="http://www.nsf.gov/funding/pgm_list.jsp?type=xcut">Cross-cutting and NSF-wide Active Funding Opportunities</a> homepage.

U.S. National Science Foundation

The Law &amp; Science Program considers proposals that address social scientific studies of law and law-like systems of rules, as wellas studies of how science and technology are applied in legal contexts.The Program is inherently interdisciplinary and multi-methodological.Successful proposals describe research that advances scientific theory and understanding of the connections between human behavior and law, legal institutions, or legal processes; or the interactions of law and basic sciences, including biology, computer and information sciences, STEM education, engineering, geosciences, and math and physical sciences.Scientific studies of law often approach law as dynamic, interacting with multiple arenas, and with the participation of multiple actors.Fields of study include many disciplines, and often address problems including, though not limited, to: Crime, Violence, and Policing Cyberspace Economic Issues Environmental Science Evidentiary Issues Forensic Science Governance and Courts Human Rights and Comparative Law Information Technology Legal and Ethical Issues related to Science Legal Decision Making Legal Mobilization and Conceptions of Justice Litigation and the Legal Profession Punishment and Corrections Regulation and Facilitation of Biotechnology (e.g., Gene Editing, Gene Testing, Synthetic Biology) and Other Emerging Sciences and Technologies Use of Science in the Legal Processes LS supports the following types of proposals: Standard Research Grants and Grants for Collaborative Research Conference Awards LS also participates in a number of specialized funding opportunities through NSF s cross-cutting and cross-directorate activities, including, for example: Faculty Early Career Development (CAREER) Program Research Experiences for Undergraduates (REU) Research at Undergraduate Institutions (RUI) Grants for Rapid Response Research (RAPID) Early-concept Grants for Exploratory Research (EAGER) For information about these and other programs, please visit the Cross-cutting and NSF-wide Active Funding Opportunities homepage.

Lemelson Foundation

Supports invention-based education, STEM learning, and inventor support programs. Focus on inspiring young inventors and commercializing socially beneficial inventions.

NEI - National Eye Institute

PROJECT SUMMARY IMPG2 is a crucial extracellular matrix protein for maintaining photoreceptor structure and function. Mutations in IMPG2 are linked to two forms of visual impairment: juvenile-onset rod-cone dystrophy and adult-onset vitelliform macular dystrophy. While no treatment currently exists, packaging IMPG2 amino acid sequences into adeno-associated viruses (AAVs) offers promise for a sight-preserving therapy. We recently generated human retinal organoid (RO) and mouse models to enable us to rapidly engineer a gene augmentation therapy for IMPG2-associated retinal dystrophy (RD). The retinal organoids (ROs), grown from either patient-derived (human) induced pluripotent stem cells (hiPSCs) or gene-edited embryonic stem cells (hESCs), recapitulate the lack of photoreceptor outer segments observed in advanced IMPG2-RD. This fully penetrant phenotype provides a biomarker for assessing functional IMPG2 expression after AAV-mediated gene transfer. Although patient-derived ROs are tractable in vitro models of clinical relevance, their use in assessing viral vector designs for gene therapy development is best complemented by in vivo assessment of safety and efficacy in animal models. Accordingly, we will assess therapeutic safety and efficacy using the Impg2-knockout (KO) model mice, as these mice exhibit gliosis, subretinal deposits, photoreceptor degeneration, retinal detachment, and reduced electroretinogram (ERG) responses that are similar to the human condition. Here, we will accelerate a preclinical program to test our central hypothesis that gene augmentation can prevent retinal pathology in an IMPG2-RD mouse model and patient-derived ROs. To lay the groundwork for a clinical IMPG2 gene therapy, we will complete the following Aims: (1) Use Impg2-KO mice to define endpoints for preclinical trials, (2) optimize a gene therapy viral vector design using Impg2-KO mice and IMPG2 patient-derived and gene-edited ROs, and (3) establish preclinical gene therapy safety and efficacy in Impg2-KO mice. Synergistically employing mouse models and human ROs will accelerate the development of a gene therapy that will meaningfully improve the lives of individuals with IMPG2-RD. More broadly, this work will demonstrate the power of a dual-model platform to advance safe and effective therapeutics with high predictive value for treating inherited retinal disorders.

NIA - National Institute on Aging

The epigenome controls cell type-specific gene expression, establishing the diversity of cell types in the human body. However, over time, the epigenome becomes dysregulated, which promotes aging. Despite the tight link between epigenetics and aging, the mechanisms that preserve the epigenome in young cells and why these mechanisms degrade over time remain poorly understood. Polycomb-mediated gene repression maintains cell identity by silencing the genes that specify other cell types. As facultative heterochromatin, Polycomb is highly dynamic during development, enabling differentiating stem cells to rapidly alter gene expression programs. However, Polycomb switches from being flexible during development to becoming a stable mechanism of repression throughout adulthood. Understanding Polycomb regulation is key to advancing our knowledge of aging, as disrupting Polycomb components alters lifespan across various organisms. How Polycomb repression is maintained in terminally differentiated cells remain unknown. However, studies in embryonic stem cells indicate that spatial organization of repressed sites is crucial, with Polycomb-repressed regions forming ultra-long-range loops to sustain silencing. While these loops were thought to be solely mediated by Polycomb complexes, preliminary work from the applicant shows that cohesin and CTCF (which facilitate long-range enhancer-promoter loops) also mediate repressive loops in embryonic stem cells. In the F99 phase of this proposal, performed at MIT, the applicant will use computational methods developed by the Mirny and Dekker labs to determine whether cohesin and CTCF-dependent looping is a broad regulatory mechanism of Polycomb repression. Aim 1.1 will identify Polycomb targets in embryonic stem cells that derepress when cohesin or CTCF is lost and Aim 1.2 will use mechanistic polymer modeling to link cohesin and CTCF’s roles in 3D looping activity to Polycomb repression. In Aim 1.3, machine learning and polymer modeling will predict how gene expression in different cell types, particularly mature hepatocytes, respond to site-directed CTCF perturbations. These insights will propel the applicant’s transition to aging research, where she will test whether enhancing cohesin activity can protect Polycomb repression in aging mouse livers (Aim 2). The K00 phase will also use cutting-edge and single-cell experimental techniques to measure genome re-organization as Polycomb becomes dysregulated during the normal aging process. In addition to training in machine learning and hepatic chromatin, the applicant will gain expertise in aging research during the F99 stage through lab visits, conference attendance, and a course on aging and its diseases. This study will advance our understanding of aging by comprehensively investigating a new mechanism of Polycomb regulation. Rejuvenating the epigenome is a promising strategy for reversing cellular aging, and this work will determine if targeting the 3D genome offers a new approach.

NIGMS - National Institute of General Medical Sciences

PROJECT SUMMARY Human mesenchymal stem cells (hMSCs) can be isolated from various adult tissues including bone marrow, fat tissues, dental pulp and synovium, and exhibit therapeutic promise for tissue repair, regeneration, and treatment of chronic immune and degenerative diseases due to their multipotent differentiation ability, trophic factor secretion, and immunomodulatory functions. However, their clinical efficacy remains limited by two barriers: (i) immune-mediated clearance of transplanted cells and (ii) poor retention and engraftment within injured tissues. Even autologous or well-matched allogeneic hMSCs are frequently recognized as “non-self” after their transplantation, leading to rapid cell loss before therapeutic benefits are realized. Overcoming immune rejection and microenvironmental insufficiency is therefore critical to advance hMSC-based therapies. Systemic immunomodulators can partially mitigate host rejection but often cause off-target effects, while conventional scaffolding like hydrogels provides structural support yet rarely influences immune responses. To address these limitations, we propose a light-activated hydrogel platform that integrates local immunomodulation with in situ scaffolding under a single, spatiotemporally defined stimulus. To achieve this objective, we will pursue two aims: 1) design, synthesis and characterization of BEPA-photocaged hydrogel complex for controlled immunomodulator and hydrogel formation, and 2) preclinical evaluation of BEPA- photocaged hydrogel complex for immune-permissive and ECM-mimetic stem cell engraftment. The proposed platform is expected to establish a “regenerative niche” that is both immune-compatible and mechanically supportive, thereby improving hMSC survival, retention, and integration. The approach is innovative in three ways: (i) a light-activated photocage enabling precise, on-demand immunomodulation, (ii) an injectable, light- triggering hydrogelator that surpasses conventional scaffolds, and (iii) a unified near-infrared (NIR) control axis that simultaneously regulates immune modulation and scaffold formation. By converting engraftment into a programmable event, this strategy provides effectively spatiotemporal control over stem cell transplantation. The anticipated outcome is a safer and more effective platform for regenerative medicine with broad translational potential.

NIDDK - National Institute of Diabetes and Digestive and Kidney Diseases

PROJECT SUMMARY Barrier epithelia face continual damage from environmental insults, and successful injury repair is crucial for organismal health. A prime case study is the one-cell-thick intestinal epithelium, which forms a leakproof bar- rier between the gut lumen and the body cavity. To replace damaged cells, the intestine mobilizes stem cells to divide rapidly; to restore intestinal form and function, these new daughter cells must also differentiate rapidly. Indeed, injury-born intestinal cells acquire their mature identity twice as fast as their normal counterparts. Using the Drosophila adult intestine, we recently discovered this injury-accelerated differentiation arises through disruption of Notch-Delta lateral inhibition circuitry that normally specifies stem versus terminal fate. During injury, many newly born cells exhibit >10x faster Notch signaling speed, which propels faster intestinal differentiation to restore the breached epithelial barrier. Yet this strategy comes with risks for long-term tissue health: For stem cells, loss of Notch-Delta feedback during injury skews daughter fates toward dead-end, ter- minal:terminal outcomes, which depletes the organ’s stem cells and culminates in stem cell exhaustion. For terminal progeny, accelerated differentiation yields provisional ‘stopgap’ cells—mature cells with digestive and barrier-forming functions but altered morphology and a supercompetitor-like transcriptomic profile. Here, we will investigate how the organ copes with these two tradeoffs. We combine physiological injury of the fly gut with in vivo live imaging and cutting-edge cell lineage tracing to elucidate how these ‘side effects’ of accelerated differentiation are managed at the organ-scale for post-injury tissue homeostasis. The fly gut com- bines conserved intestinal cell lineages, fate signals, and digestive physiology with supreme experimental trac- tability: A single Notch receptor and Delta ligand, an unparalleled wealth of genetic tools, and long-term in vivo live imaging—pioneered by our lab—provide the technical bases for deep mechanistic investigation. Leveraging these strengths, in Aim 1 we will define how some, ‘escaper’ stem cells persist after injury, de- spite disrupted Notch-Delta feedback that should force all cells to differentiate. We will test if escaper stem cells inherit an intracellular Notch inhibitor, autonomously override how Notch and Delta interact, or lose con- tact with their signaling partners via injury-induced epithelial fluidization. In Aim 2, we will ascertain the time- evolution and function of stopgap cells during and after injury. We will determine their ultimate fates in the tis- sue during recovery, e.g., they may evolve into normal cells, arrest in an abnormal state, or simply be shed. We will parse these scenarios using longitudinal live imaging, whole-population analyses, and single-cell tran- scriptomics. Finally, we will examine how stopgap cells shape the tissue post-injury by ‘purging’ unfit, toxin- exposed cells during injury or by exerting selective pressure on new cells during post-injury recovery. By probing these lineage tradeoffs of accelerated cell differentiation during injury, our work will suggest new strategies to promote intestinal regeneration and combat chronic intestinal disease.

NCI - National Cancer Institute

Abstract Breast cancer immunotherapy efficacy is still very limited. Thus, there is a pressing need to understand molecular determinants enabling clinically distinct breast cancers to suppress antitumor immune response and harness these mechanisms for novel treatments. Emerging evidence suggests that oncogenic mutations can directly adversely affect tumor immune responses. However, such functions for vast majority of breast cancer mutations have not been explored. In this proposal, we focus on aggressive breast tumors carrying combinations of most frequent breast cancer driver mutations, as a proof of principle of a new and customizable platform of clinically relevant cancer models to understand immunosuppressive mechanisms. MLL3 (also known as KMT2C), encoding a histone methyltransferase, is a novel tumor suppressor in various human cancers. MLL3 is frequently inactivated by gene deletions or truncating point mutations, with especially high rate in breast cancers (up to 24%). Additionally, MLL3 mutant cancers have significantly worse outcomes compared to MLL3 wild type cancers. We have developed a novel method for rapidly generating genetically engineered mouse models (GEMMs) by efficiently expanding and “custom genome editing” mouse mammary stem cells (MaSCs) in culture and then using these MaSCs to regenerate genetically engineered mammary glands in syngeneic immunocompetent mice. Using this model of MLL3 deletion in conjunction with constitutive activation of PI3-kinase (PI3KCA, ~60% of MLL3 mutant tumors in patients are also PI3KCA) and inactivation of p53 (these three mutations altogether account for the most frequent combinations of cancer driver mutations in human breast cancers), we found that the loss of Mll3 promotes early infiltration of Foxp3+ regulatory T (Treg) cells and their further expansion and differentiation to a highly suppressive phenotype, leading to faster tumor immune escape in primary tumors and at metastatic sites. Monoclonal antibody targeting of specific immune receptors highly expressed on tumor-infiltrating Treg cells show remarkable efficacy in inhibiting tumor initiation and growth. Based on these findings, we next propose to investigate how MLL3 loss mechanistically activates HIF1 and harness the understanding to develop therapeutic interventions applicable to aggressive human breast cancers. We will explore underlying mechanisms of Treg cell differentiation into highly suppressive effector Treg cells in the tumors, mediated by both extracellular cues and cell-intrinsic regulators. Thus, in addition to uncovering new mechanisms by which major breast cancer drivers favor early immune escape, the power of our approach can be easily extended to test the tumor promoting effect of any breast cancer mutations, singly or in combination. The mechanistic investigations of these complex tumor-immune system interactions in vivo require the use of similarly complex in vivo models that closely recapitulate tumor development in an immunocompetent host environment in which tumors arise. To our knowledge, mice represent the most cost effective and best tractable mammalian models available to us.

NIGMS - National Institute of General Medical Sciences

PROJECT SUMMARY/ABSTRACT Adult stem cells are progenitor cells capable of tissue regeneration during life through the ability to both self- renewal and to produce specialized cells upon division. Stem cells reside in microenvironments called “niches” that integrate systemic cues and provide signals for stem cell maintenance. Over the years, the use of the stem cell systems present in Drosophila melanogaster has revealed mechanisms that control stem cell niches in homeostasis and pathology. Recently, a model has emerged pointing to a strong conserved correlation between lipid accumulation and stem cell loss. Given the power of Drosophila genetics, the readily accessible molecular tools, the well-characterized stem and niche cell populations, and the high degree of evolutionary conservation in metabolic genes, the fly testis niche is an ideal model for the intersectional study of metabolism and stem cell biology in physiological and pathophysiological conditions. Our long-term goal is to understand how changes in lipid metabolism affect stem cell niche homeostasis. The PI’s published works build a model where the ectopic accumulation of lipids in the fly testis niche is detrimental to stem cell function. Excess lipid accumulation in stem cells led to their loss through differentiation. Accordingly, lipid accumulation has been shown to be detrimental to stem cell maintenance across species. The overall objective of this proposal is to understand mechanistically how the stem cell niche is affected by conditions that trigger ectopic accumulation of lipids. Preliminary data in this proposal show that niche (hub) cells are also sensitive to lipid accumulation, and that multiple mechanisms are likely at play to control lipid levels in the testis stem cell niche. Of note, preliminary data in this proposal show for the first time that lipid metabolism controls somatic cell fate in the testis by inducing conversion between niche and somatic stem cells. Hence, our central hypothesis is that lipid accumulation promotes loss of stem cell niche homeostasis. We will test this hypothesis through three specific aims: 1) determining how microenvironmental stiffness impacts lipid anabolism and niche homeostasis; 2) characterizing the role of apolipoproteins in fat-transporting and stem cell maintenance; and 3) investigating the role of lactate transport (a precursor in lipogenesis) in niche and stem cells. The merit of this study relies on its novelty – showing that changes in lipid metabolism can promote the conversion between a niche and a stem cell – and on the generation of a useful paradigm for testing how pathophysiological changes in lipid metabolism yield in loss of stem cell niches. Given the high incidence of metabolic disorders in the population, understanding how lipid accumulation affects stem cell niches is pivotal for the development of novel stem cell-based therapies, especially those targeting metabolic disorders. The proposed studies will also strengthen the research environment at the University of Louisville, providing opportunities for the training of postdoctoral fellows, graduate and undergraduate research assistants in the laboratory.

NIGMS - National Institute of General Medical Sciences

Abstract The overall theme of my research program is to understand how enhancers and transcription machineries organize in the 3D genome to regulate the target gene expression. We plan to address this problem from two aspects: 1) at the single-cell level, we will characterize the function and dynamics of transcriptional condensates in mouse embryonic stem cells and investigate the specificity of the condensate, the formation and dissolution mechanisms, and their activity regulated by genome architecture; 2) at the developmental level, we will study the mechanism of how transcriptional condensates achieve differential regulation of target genes during development using the beta-globin regulation in mouse erythroleukemia cells as a model. In the past years, we have made significant contributions in the first direction, and we excited to extend our research to a new direction. New methods were developed and insights generated from these studies form the foundation of this proposal. Theme1: In eukaryotes, RNA Pol II, Mediator and transcription activators form transcription-dependent condensates in live mouse embryonic stem cells and enhance super-enhancer controlled gene bursting in a proximity-dependent manner. However, molecular mechanisms of how condensates function on gene bursting within the 3D genome is not fully resolved. The long-term goal of this theme is to characterize the condensate specificity, to investigate their mechanisms of formation and regulation by enhancers/enhancer RNAs, and to develop a full mechanistic understanding of how they interact with their target regulatory elements. In the next five years, we plan to 1) test whether transcriptional condensate can co-activate a reporter gene with different promoter specificity; 2) develop a labeling strategy to image and study the roles of enhancer RNAs in condensate function; 3) selectively manipulate chromosomal interactions and test how condensate-gene interactions and gene bursting change. Theme 2: The dynamics and functions of transcriptional condensate have not been explored extensively due to their close-to-diffraction-limit size. The long-term goal of this theme is to characterize the composition, dynamics and functions of transcriptional condensates and their effect on beta-globin gene switching during erythroid differentiation, aiming to test the function of condensates across different systems. In the next five years, we plan to: 1) test whether and how transcriptional condensate dynamically controls beta-globin gene bursting; 2) develop a method to probe the condensate composition and its changes during erythroid differentiation. We envision that our study will provide significant insights and open new research directions on gene regulation.