University Of Chicago

Fonds de recherche du Québec – Société et culture · FY 2023-2024 · 2023-04

Volet: Bourses postdoctorales; Domaine: Arts, littérature et société; Objet: Arts et traditions culturelles; Objet: Échanges culturels; Application: Structures et relations sociales; Application: Culture; Mots-clés: BRITISH EMPIRE, COLONIALISM, SOUTH ASIAN ART HISTORY, BRITISH ART HISTORY, ART MARKET, MUSEUMS

Fonds de recherche du Québec – Nature et technologies · FY 2023-2024 · 2023-04

Volet: Bourses de maîtrise en recherche; Domaine: Nature et interactions de la matière; Objet: Phénomènes quantiques; Objet: Mathématique informatique; Application: Sciences et technologies; Application: Fondements et avancement des connaissances; Mots-clés: MATIERE CONDENSEE, INFORMATION QUANTIQUE, INTRICATION, ROBUSTESSE, TRANSITIONS DE PHASE QUANTIQUES, NEGATIVITE LOGARITHMIQUE

NIH Research Projects · FY 2026 · 2023-03

The purpose of the NIDDK R25 Research Education Training Program, DULCE (Diabetes InqUiry through a Learning Collaborative Experience) at the University of Chicago Pritzker School of Medicine is to inspire medical students to pursue careers in diabetes mellitus (DM) research through exposure to the full continuum from basic science to translational DM research and the interdisciplinary collaboration needed to care for patients with DM.  Given the increasing number of Americans with diabetes mellitus (DM), it is imperative that our physician-scientist work force is well prepared to address the needs of this growing population and keep pace with DM innovation. The University of Chicago is home to two strong DM research centers whose investigators currently have $31.5 million in grant funding from NIDDK and represent research that spans basic science, clinical translational, community based, and policy research. UC also provides high caliber clinical care through the Kovler Diabetes Center and other highly ranked clinical programs. The Pritzker School of Medicine (PSOM) has the potential to be a leader in training medical students in DM research;;42% of incoming PSOM students express interest in conducting DM research. While the PSOM has a track record in training students in research, we lack a program focused on DM. To address the gaps in the current physician scientist workforce and meet the needs of our current students’ interest in DM research, we will develop, implement, and evaluate DULCE. We propose this novel longitudinal research education program for eight medical students per year during their first and second year of medical school consisting of a short-term basic science, clinical, or community health research experience combined with a structured didactic seminar series and interdisciplinary clinical shadowing experiences. DULCE will provide students with concrete experiences in DM research and clinical shadowing, reflective observation through regular meetings with peers and physician scientists, learning of new theories through participation in multidisciplinary clinical and research seminars, and application of new concepts through an intensive mentored research project. DULCE’s evaluation will be based on assessments of students’ research self-efficacy, their attitudes towards DM research, their exposure to the roles of interdisciplinary health professions in DM care, and their ability to contribute to a research project. We will also assess their interest in pursuing a research career in DM and their productivity in a future career in DM research through tracking of conference presentations and publications. Our goal through DULCE is to provide PSOM students with a better understanding of the spectrum of DM research and interdisciplinary collaboration, inspiring them to pursue careers in DM research.

NIH Research Projects · FY 2025 · 2023-03

PROJECT SUMMARY Adjuvants included as a component of a vaccine have a major impact on vaccine efficacy via modulating and prolonging host immune responses. While vaccines are the most effective way to prevent and control infectious diseases, many pathogens that significantly impact human health remain without an effective vaccine. For example, one-fourth of the world’s population is latently infected with Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB)1, a leading infectious disease killer in the world. The currently licensed TB vaccine, M. bovis BCG (BCG) provides variable protective efficacy (0-80%) across the world2-4. In the absence of clear correlates of protection for TB vaccines, it is imperative that we explore new and effective approaches to target broad T cell responses for vaccine-induced immunity against TB. In the current proposal, we hypothesize that use of new adjuvants that drive Th1 responses (UM-1007, a novel TLR7/8 agonist) and Th17 responses (UM-1098, a novel Mincle agonist) will significantly augment vaccine-induced protection against TB. Thus, during the R61 phase of the proposal, we will optimize the use of these novel Th1- and Th17-inducing adjuvanted vaccine platforms and conduct proof-of-principle studies including detailed assessment of immunogenicity, antigen optimization, identification of prime-boost strategies that enhance lung Th1- and Th17- vaccine responses (Specific Aim 1), and test if these novel Th1- and Th17-inducing TB vaccine candidates will protect in mouse challenge models of TB (Specific Aim 2). During the R33 phase we will test the optimized Th1- and Th17-inducing TB candidate vaccines for immunogenicity, safety and protection in an aerosol challenge model in macaques (Specific Aim 3). We also propose that we will meet the following milestones. R61 phase. Milestone 1 (Month 12): Antigen selection for an optimized IM vaccine producing a Th1 response (UM-1007:TB) and a Th17 response (UM-1098:TB). Milestone 2 (Month 18): Identification of the best prime-boost strategy for a TB vaccine inducing maximal lung Th1 responses (UM-1007:TB) and Th17 responses (UM-1098:TB). Milestone 3 (Month 33): Testing efficacy of optimized UM-1007:TB and UM-1098:TB candidates against drug susceptible and drug resistant Mtb and in genetically diverse Mtb-infected mice. Milestone 4 (Month 36): Submission of R33 transition package. R33 phase. Milestone 5 (Month 60): Immunogenicity, safety and efficacy studies of optimized UM-1007:TB and UM-1098:TB vaccines in macaques and in-depth characterization of vaccine-induced immune responses. Milestone 6 (Month 60): Final selection of newly discovered Mtb vaccine. Thus, the work proposed in this grant will result in a novel, first-of-kind rationally designed Th1- and/or Th17- inducing TB vaccine for use in humans in the near future.

NIH Research Projects · FY 2026 · 2023-03

PROJECT SUMMARY / ABSTRACT Collective migration of epithelial cells plays central roles in morphogenesis, intestinal turnover, wound repair, and metastasis. Epithelial cells use the same migration machinery as individual cells. For an epithelial sheet to migrate, however, this machinery must become globally aligned across the tissue plane. Determining how this tissue-level polarization is achieved is a central goal of the collective migration field. We study a rotational form of epithelial migration that occurs when the tissue is confined to a circular or spherical geometry. Rotational migrations differ from other epithelial migrations in two ways. First, external cues like empty space or chemo- tactic signals are not available to guide tissue polarization. Instead, the cells must rely solely on local cell-cell interactions to achieve this state. How cells self-organize for rotational migration is unknown. Second, there is no net tissue movement, which raises the question of why these migrations occur. Rotation promotes the assembly of the basement membrane extracellular matrix that lines the tissue’s basal surface; it can even create highly structured basement membranes that direct organ morphogenesis. However, how rotation impacts basement membrane assembly is poorly understood. Notably, recent work has shown that epithelial rotation may contribute to human organ development, as the spherical alveoli of mammary organoids rotate as they form despite being connected to a central ductwork. My NIGMS-funded research has two goals: (1) to define the local cell-cell interactions that allow epithelial cells self-organize for rotational migration, and (2) to determine how rotation structures the basement membrane. To this end, we are studying a rotational migration that occurs in the follicular epithelium of the Drosophila. In recent years, we used this model to provide the first insight into the local cell-cell interactions that polarize an epithelium for rotational migration by identifying a novel planar signaling system that mediates this process. We also showed that rotation works with new protein secretion to create fibrils in the basement membrane that control tissue shape. Through the MIRA program, we will dig deeper into both mechanisms. We will determine how the planar signaling system allows the follicle cells to break symmetry and initiate migration and how the signaling works at molecular level - both in terms of how the proteins interact with one another and with the migration machinery. We will also explore two mechanisms by which mechanical forces imparted by rotational migration are likely to influence basement membrane assembly. This work will reveal new guiding principles for how tissue-level order can emerge from local cell-cell interactions. Moreover, because basement membranes are central to most organs and defects in their assembly underly muscular dystrophy, nephropathy, skin blistering, and stroke, anything we learn about this poorly understood process will have broad impact.

NIH Research Projects · FY 2025 · 2023-03

PROJECT ABSTRACT Chronic Obstructive Pulmonary Disease (COPD) affects more than 16 million US adults, many of whom experience high rates of acute care revisits (emergency department and hospital) after initial COPD hospitalization. These frequent exacerbations, often due to transition of care (TOC) failures, lead to lung function decline and earlier mortality. Large multi-site studies are needed to address ongoing gaps in fundamental effectiveness and implementation data, before widescale implementation of effective COPD TOC programs and successful reduction of revisits. My current NHLBI R01 aims to address these gaps in effectiveness and implementation by testing COPD TOC programs across 20 diverse US hospitals to improve health care delivery and outcomes for patients with COPD. My R01 simultaneously uses implementation science (IS) and human-centered design (HCD) methods. While these methods are rigorous and complimentary, integrating these qualitative approaches in a prospective manner could optimize their potential to impact the design and implementation of interventions in order to obtain data and ultimately inform care transformation. Further, standardizing metrics for evaluating and reporting COPD TOC program and revisit data could significantly improve outcome comparisons between COPD TOC programs and studies. The K24 will allow me to address these methodology gaps by building upon my R01 research, mentoring program, and infrastructure to develop, test, and disseminate integrated IS and HCD qualitative and quantitative methods for use in Patient Oriented Research. I will collaborate with my mentees and R01 research team experts in IS and HCD methods to develop an integrated approach named "CHIMERA"; additionally, I will collaboratively develop harmonized reporting metrics called "SMART COPD". I will complete these aims by working with my mentees to develop, test, and disseminate the "CHIMERA" and "SMART COPD" tools. CHIMERA will use a step-wise approach to identify HCD and IS domains and methods, followed by the use of logic models and concept mapping of stakeholder assessments from my R01 Aim 1 to develop a CHIMERA “cross walk” methodology that integrates both methods. CHIMERA will be tested in conjunction with my R01 Aims 2 and 3, implementation and post-implementation. The SMART COPD metrics will be developed using systematic literature review and a modified Delphi approach, in which our R01 team will identify and prioritize COPD TOC Program metrics to standardize reporting. This K24 will allow me to provide mentoring opportunities to my mentees through development of novel research tools. The enriched research program of my K24 entitled: "SMART POR: Supporting and Mentoring Across Respiratory Topics in Patient Oriented Research" will also provide ample opportunities for me to support current and future research mentees to implement research and drive investigator-based careers.

NIH Research Projects · FY 2026 · 2023-03

PROJECT SUMMARY Until recently, it was thought that our memories occur as a reactivation of perception – reconjuring the same visual information and neural patterns (dubbed the sensory reinstatement hypothesis; e.g., Buckner & Wheeler, 2001). However, in recent work, we observed surprising differences between perception and memory, with separate brain voxels and patterns involved in each process (Bainbridge et al., 2021b). We observed that memory representations in the brain lose rich high-level visual information, and that the neurons sensitive to memory are anterior to those sensitive to perception. Here, we hypothesize that memories become more semanticized representations of perception, resulting in an anterior neural shift towards more semantic regions (Popham et al., 2021); we dub this the semantic transformation hypothesis. This hypothesis runs counter to a recent proposal that memory and perception are supported by two separate yet adjacent networks (the separate networks hypothesis, Steel et al., 2021). By testing these three hypotheses in conjunction, we will be able to identify the specific neural mechanisms that support visual memory and its relation to visual perception. Crucially, this project has critical importance for understanding memory disorders such as Alzheimer’s Disease, for which recent work has shown that visual and semantic information in an image can predict its eventual memory fate in a patient (Bainbridge et al., 2019a; Grande et al., 2021). Towards this question, we utilize innovative methods pioneered by our laboratory, including a direct read-out of a participant’s perception and memory through drawings that are quantified through online crowd- sourcing (Bainbridge et al., 2019a; Bainbridge, 2021). In Aim 1, we use drawings to identify the direct influences of visual and semantic information on visual memory representations. In Aim 2, we use representational similarity analyses (Bainbridge et al., 2021b) combined with functional magnetic resonance imaging to test the roles of visual and semantic information on memory patterns in the brain. Importantly, in addition to testing typical individuals, we also study the newly characterized phenomenon of congenital aphantasia (2-5% prevalence), which describes individuals with no visual memory yet intact visual perception and semantic memory (Zeman et al., 2015; Keogh & Pearson, 2018). Aphantasia serves as natural “knock- out” case to observe behavioral and neural differences between perception and memory, and our laboratory is the only laboratory in the U.S. to have studied the condition thus far (Bainbridge et al., 2021a). Armed with this unique population, in Aim 3, we will identify the neural substrates that underlie their dissociation of visual memory and vision, and test the role of semantic vs. visual content on memory. In sum, this study promises to resolve our understanding of how what we see becomes what we remember, by identifying the causes of these striking differences observed in the brain between perception and memory.

NIH Research Projects · FY 2026 · 2023-03

Over the past 50 years, the field of biophysical chemistry has learned an enormous amount about the relationships between the function, dynamics and folding of soluble proteins. The next stage in biological science involves addressing more complex problems on more challenging systems using increasingly sophisticated approaches augmented by computational methods. The Sosnick lab has followed this path. We are conducting studies of membrane proteins, condensates and disordered proteins. Our ability to address these topics arises in part from our expertise in the folding of soluble and more recently, membrane proteins. We build on this experience and knowledge to advance new or improve existing methods. The proposed research relies heavily on hydrogen exchange (HX), a method developed to study folding and dynamics yet possessing broad transferability to many methods. We plan to continue this approach and study the metabolite transport across a bilayer, stress-induced phase separation, and properties of disordered proteins in addition to membrane protein folding. Many of the projects are collaborative, leveraging our skills and interests with those of other labs, which further supports the value of our studies.

NIH Research Projects · FY 2026 · 2023-03

Project Summary: Colorectal cancer (CRC) is the second leading cause of cancer-related deaths in the U.S. The majority of CRC patients have distant or regional metastasis and a poor prognosis. Although immune checkpoint blockade (ICB) has demonstrated favorable responses and survival benefits for mCRC patients with mismatch repair (MMR)-deficient or microsatellite instability (MSI)-high tumors, it does not benefit approximately 95% of mCRC patients who have MMR-proficient (pMMR) or MSI-low lesions. There is an urgent need for methods that can sensitize pMMR/MSI-low CRC, improve recognition and presentation of tumor-associated antigens, and activate T-cell proliferation and responses for synergistic combination with ICB to overcome current limitations in clinical care for mCRC patients. We have pioneered the development of nanoscale coordination polymers (NCPs) for cancer therapy. Formed by coordination polymerization between metal ions and polydentate ligands, NCPs preferentially accumulate in tumor tissues by taking advantage of the enhanced permeability and retention effect and possess several advantages over existing nanotherapeutics. The long-term goal of our research is to establish a new treatment paradigm for metastatic colorectal cancer through the development and characterization of NCPs that can be delivered systemically. We have developed OX/SN38 NCP with a hydrophilic oxaliplatin (OX) prodrug in the core and a hydrophobic SN38 prodrug on the shell. Tumor-targeted and -activated OX/SN38 demonstrated potent anticancer effect and synergized with an anti-PD-L1 antibody (αPD-L1) for strong chemo-immunotherapy in CRC models. We have also developed a robust NCP for the co-delivery of OX and 2’,3’-cyclic GMP–AMP (cGAMP) agonist of stimulator of interferon genes (STING) to tumors. OX/cGAMP significantly prolonged the half-life of cGAMP in circulation and disrupted tumor vasculatures to enhance tumor accumulation. The overall goal of the proposed studies is to develop a tumor-targeted core-shell NCP, OX/CDN/Chol-D, through the optimization of CDN and cholesterol-conjugated drug (Chol-D) separately, for the co-delivery of OX and Chol-D to cause tumor immunogenic cell death (ICD) and the release of CDN for STING activation in the tumor microenvironment. We will elucidate the mechanisms of enhanced drug delivery to tumors via LDLR- mediated endocytosis and tumor vasculature disruption by OX/CDN/Chol-D and evaluate its anticancer efficacy alone and in combination with αPD-L1 in multiple CRC models. By creating an immunogenic tumor microenvironment, activating STING, and eliciting T-cell mediated cytotoxicity, OX/CDN/Chol-D promises to turn immunologically “cold” CRC tumors “hot” for synergistic combination with ICB to improve immunotherapy of mCRC. Our close collaborations on this multidisciplinary project promise to identify a novel tri-modality nanomedicine for clinical translation to treat mCRC patients with a poor prognosis.

NIH Research Projects · FY 2026 · 2023-03

The overarching goal of our laboratory is to understand how cells process signals and communicate robustly in complex environments. We combine experiments, modeling, and development of technologies for high- throughput single-cell analysis. Here, we build upon two grants that proposed fundamental (R01GM128042) and technological (R01GM127527) studies of immune signaling. Our goals are integrated around two key questions: How do cells encode information into signaling molecules? Environmental inputs are detected by sentinel cells, which in response produce cytokines. Recent studies led to the hypothesis that cytokine dynamics encode information from the environment. Determining how pathogen and stress inputs are encoded is crucial for understanding infection, autoimmunity, sepsis and cancer. We will use single-cell protein production/secretion assays and live-cell analysis to answer key questions including: How do cells exposed to multiple sequential stimuli encode the memory of prior stimuli? How does cellular density and coordination influence cytokine responses? What are the mechanisms of cytokine production variability by myeloid or epithelial cells, and what causes excessive cytokine production/secretion? Technologies to analyze single-cell protein production/secretion: We will realize a method for simultaneous measurement of single-cell expressed proteins, protein complexes and mRNA using single-cell sequencing readout. High-throughput microfluidic technologies for time-dependent measurement of proteins secreted by single live-cells will be developed. We will also develop microfluidic co-culture systems for creating controlled dynamic microenvironments. How do cells process combinatorial/dynamic signals? Signals generated by sentinel cells are processed by transcriptional pathways. NF-κB, an inflammatory pathway that controls responses to many signals, is a prime example of a system that creates fine-tuned responses. We will study NF-κB as a model system to answer key questions in signal processing in single-cells: How does NF-κB process combinatorial signals? During infection, immunity and stress, cells are exposed to combinations and temporal sequences of multiple cytokines. How NF- κB processes such inputs is not understood. How does NF-κB dynamics regulate gene expression in space and time? While much attention has been given to temporal characteristics of signaling, little is known about how NF- κB decodes signals propagating over different spatial scales. How do single cells/populations respond to dynamic (increasing, decreasing, oscillating) inputs? High-throughput live-cell analysis technologies: To better answer these and other general questions in signaling, we will develop broadly applicable live-cell analysis technologies: We will develop microfluidic systems that culture, track and analyze single-cells and populations under predetermined dynamic/combinatorial signals. We will develop microfluidic technologies for spatial analysis of live cells and cell signaling events. We will develop computational/statistical methods for automated analysis for image segmentation, cell/organoid tracking, and prediction of cellular outcomes.

NIH Research Projects · FY 2026 · 2023-02

Accurate and multiplexed characterization of proteins is essential to basic and clinical studies in immunity, infection, development, and cancer. Many processes in immune development, signal activation, and drug resistance are driven by a small subset of cells and variable activation of signaling pathways, necessitating single-cell measurements. Currently, there is high precision and throughput in measuring DNA/RNA in single cells, however a major technological gap exists in the measurement of proteins and especially their complexes in individual cells. High-throughput methods combining simultaneous measurement of proteins, complexes and mRNA are needed to better understand and model individual cellular responses, and to discover new cell states and functions. Our proposal has two, equally important, and synergistic goals: a) optimize/adapt a broadly applicable and practical technology that simultaneously measures proteins, protein-complexes and mRNA in thousands of individual cells (Aim 1), and b) study several key hypotheses on the function and evolution of signaling networks during immune development (Aims 2 and 3). Our technology, called Intracellular Proximity- Sequencing (iProx-seq), uses DNA barcoded proximity probes and single-cell sequencing for multiplexed measurement of proteins and their complexes. The number of protein complexes measured by iProx-seq scales quadratically: Targeting 100 proteins will enable the measurement of 5500 potential protein complexes in each cell. Protein quantification by sequencing has the additional benefit of transcriptome-wide mRNA measurements in the same cell, all using a robust and widely used sequencing pipeline. Extensive preliminary data we present demonstrated the feasibility of our entire technical approach and mechanistic studies. We will combine iProx-seq, live cell imaging and mathematical modeling and study key hypotheses in the differentiation of hematopoietic stem cells (HSCs) and B cells in the germinal center (see Aims 2 and 3). We will measure signaling receptors, adaptor proteins, transcription factors, cytokines, kinases, and protein modifications, and comprehensively characterize immune signaling networks NF-κB, MAPK, PI3K and IRF3 in single human and mouse HSCs, monocyte derived macrophages, granulocyte-monocyte progenitors, and germinal center B cells. Specific questions we will answer include: How do changes in receptor levels, receptor- coreceptor complexes, and intracellular complex formation explain single macrophage sensitivity to inflammatory TLR signals? How does the developmental remodeling of protein networks NF-κB, MAPK, PI3K and IRF3 regulate signal specificity across the hematopoietic lineage? What are the distinct proteomic and signaling states in the germinal center, and how do protein networks regulate the differentiation of B cells? Our proposal will result in a powerful and practical single-cell analysis technology and improved insight on the function and evolution of protein networks in immunity. Our results will make significant impact into the understanding of immune development, immune activation, and emergence of drug resistance.

NIH Research Projects · FY 2026 · 2023-02

Project Abstract Cellular communication is essential for the development of all multicellular organisms, and is a key phenomenon that is disrupted in many human diseases. Cell-surface receptors mediate cellular communication and are the targets for 50% of FDA-approved drugs, highlighting their disease relevance and druggability. However, many cell-surface receptors remain understudied and undrugged. The research proposed in this application is focused on two families of such receptors, adhesion G Protein-Coupled Receptors (aGPCRs), and teneurins. The structures, mechanisms of action and disease relevance of these two receptor families remain largely unknown; as their large size and complex biochemical nature has made them difficult to study. Recent genetic studies revealed that aGPCRs and teneurins have essential roles in development of the nervous system, skeletal system, and heart. These receptors are linked to diseases including cancer, developmental disorders, and brain malformations, raising an urgent need for mechanistic studies on aGPCRs and teneurins. My research program aims to elucidate the molecular mechanisms by which these receptors are activated, and to develop new tools to modulate their activity against relevant diseases. The research proposed in this application involves an interdisciplinary approach, integrating structural studies of these receptors and their ligands, biochemical and biophysical assays, protein engineering approaches, and functional assays. This research will build on our previous successes using this approach, which has yielded many three-dimensional structures of aGPCRs and teneurins, has revealed crucial mechanistic concepts in the field, and has allowed a better understanding of the mechanisms of action of these receptors. A major revelation from our work is that the large extracellular regions of aGPCRs and teneurins are directly involved in regulating receptor function. Despite these advances, fundamental questions remain unanswered. The ultimate goals of this proposal are to reveal how signal transduction is mediated within the domains of these large receptors, what exactly activates them under physiological conditions, how their evolution from early organisms to higher eukaryotes have changed and diversified their critical functions, and finally how, at the molecular level, we can inhibit or activate these receptors using synthetic ligands. Our structural studies will be complemented with mutagenesis, signaling assays, the use of synthetic binders to understand how the different components of these receptors control receptor function, and physiological analyses performed by collaborators that will test the relevance of the structural and functional studies. We expect that this research will provide critical insights into the mechanistic details of aGPCR and teneurin function that will be highly informative for the development of future therapeutics; we will also produce potent and selective synthetic ligands which can serve as tools for the scientific community to study aGPCRs and teneurins.

NIH Research Projects · FY 2025 · 2023-02

PROJECT SUMMARY/ABSTRACT Approximately one-fourth of the world's population is latently infected (LTBI) with Mycobacterium tuberculosis (Mtb) and those infected have a 10% lifetime risk of developing clinical pulmonary TB (PTB) disease over their lifetime. Global efforts to combat TB are hampered by the emergence of extensively drug-resistant and multidrug- resistant strains and the variable efficacy of the currently available vaccine, M. bovis Bacille Calmette Guerin. Unfortunately, the immune mechanisms that govern progression from LTBI to PTB are poorly defined, preventing rational design of treatments or vaccines that stimulate immune control of TB. Thus, there is an urgent need to better understand the immune parameters that contribute to Mtb control. The tubercle granuloma is a hallmark of TB and is important for the immune control of Mtb infection. However, not all granulomas effectively control TB, as they are seen both during LTBI and during TB reactivation (TB-R) and PTB. During the previous R01 funding period, we used human TB samples and non-human primate (NHP) and mouse models of TB to demonstrate an unequivocal protective role for inducible bronchus-associated lymphoid tissue (iBALT) containing clearly defined B-cell follicles during both primary and vaccine-induced responses to Mtb infection. Additionally, we determined that a dominant feature of granulomas during PTB is the accumulation of neutrophils that produce proinflammatory molecules such as S100A8/A9 proteins and limit iBALT formation. Our long-term goal is to develop clinical strategies to restrain Mtb growth and prevent progression from LTBI to TB disease. Based on work from this funded R01, the objective of this R01 renewal is to identify the roles of granuloma- associated B cells and neutrophils in TB infection and disease progression and to determine if iBALT is an effective target for preventing TB progression. Towards this overall goal, we propose to use both small animal and large animal models to address the following specific aims: In Specific Aim 1 we will investigate the role of iBALT-residing B cells in protecting against TB; In Specific Aim 2, we will evaluate the role of neutrophilic proteins such as S100A8/A9 in limiting iBALT during PTB, and to determine if targeting S100A8/A9 pathways will reduce TB disease and improve Mtb control; Finally, in Specific Aim 3, we will determine if iBALT structures can be targeted to prevent TB progression in Mtb-infected hosts. The knowledge to be gained from these studies is significant, as it will directly advance the development of new therapeutics and vaccine strategies for limiting TB-R and progression to TB.

NIH Research Projects · FY 2026 · 2023-02

The Mid-America CTSA Consortium (MACC) submits this renewal application to continue as a Regional Clinical Center (RCC) for the “Strategies to Innovate EmeRgENcy Care Clinical Trials” (SIREN) Network (RFA-NS-22-015). Because Dr. Aufderheide (previous Contact PI at the Medical College of Wisconsin [MCW] Hub) has announced his retirement on January 1, 2024, and because his replacement is unknown at the time of grant submission, we propose to transfer the MACC Hub from MCW to the University of Chicago Medicine (UCM) and Contact PI status to David Beiser, MD, MS (UCM) proactively. As recommended by the NIH Project Officer, we are submitting this application as a Renewal, rather than a New application, providing justification for this transfer. The structure of MACC otherwise remains unchanged. Methods: MACC has a proven track record of exemplary patient accrual performance. Despite not having a site within its catchment capable of participating in the HOBIT Trial, MACC is currently tied for 5th highest enroller among 11 Hubs, enrolling 116 subjects (10.0% of total) including 35 subjects (3rd highest) in BOOST-3, 38 subjects (5th highest) in ICECAP, and 43 subjects (6th highest) in C3PO. Patient retention was 98.3% (114/116; 4th highest). This renewal application adds a strategic focus on even greater patient accrual by: 1) investing $10,000/year infrastructure funding in research coordinator support at all 6 MACC sites, 2) a Patient Accrual Liaison (PAL) to proactively identify accrual barriers, 3) rapidly resolving barriers by leveraging CTSA Collaborative Council support, 4) incorporating a Clinical Trial Practicum (entering SIREN patients) into the Clinical Trials Fellowship (CTF) to augment patient accrual, 5) adding 7 new ED expansion sites and 196,455 potential patients/year, and 6) securing an additional $1.71 Million of institutional funds for SIREN. The UCM research team, experienced in supervising Spokes, and the Center for Health and the Social Sciences, experienced in subcontract management, are strong assets. The CTF provides targeted curriculum, over 20% protected time, a Clinical Trial Practicum, K Award submission post completion, and an additional $50,000 institutional funding to train 3, 2- year fellows. All MACC sites agree to continue Reliance and Master Contract Agreements. With a Hub, 5 Spokes, 3 EMS, 4 helicopter systems, catchment of 9,931,200 people, 110,598 potential patients/year, 190 multi- disciplinary collaborators, expansion capacity of 23 adult, 5 pediatric hospitals, and 2 EMS systems, 46 research personnel, 24/7 coverage at all sites, a strategic patient accrual plan, and proven performance, MACC is well- positioned to achieve SIREN goals. Specific Aims. MACC aims to: 1) Meet/exceed recruitment goals of at least 100 subjects per year, including diverse/underserved populations, 2) Achieve all study milestones and exemplary clinical trial execution, 3) Engage a wide spectrum of investigators, 4) Mentor 3 junior faculty investigators to successful academic careers in emergency clinical trial research, 5) Contribute significantly to SIREN governance, and 6) Promote dissemination of SIREN study findings and stimulate SIREN grant applications.

NIH Research Projects · FY 2025 · 2023-01

PROJECT SUMMARY/ABSTRACT The ability of fibroblasts to synthesize extracellular matrix (ECM) proteins is critical for wound healing; however, this program is coopted by cancer cells in multiple solid tumors, resulting in the formation of cancer-associated fibroblasts (CAFs) that drive a desmoplastic response which contributes to cancer progression and promotes therapy resistance. Collagen is the most abundant protein in the ECM and has a unique amino acid composition such that up to 25% of its sequence is represented by proline. Proline is a limiting metabolite for collagen synthesis in CAFs, and CAFs promote proline biosynthesis by increasing mitochondrial glutaminolysis and directing the resulting glutamate into the proline biosynthetic pathway instead of its utilization as an anaplerotic substrate for the TCA cycle. However, the mechanism directing the differential utilization of glutamine for proline or the TCA cycle is not understood. Besides, glutamine is depleted in many solid tumors such as pancreatic ductal adenocarcinoma (PDAC), but paradoxically, collagen is highly abundant in PDAC. The objective of the present proposal is to gain a better understanding of the unique metabolic requirements for collagen production in CAFs and thereby identify novel targets to reduce tumor desmoplasia. The proposed studies are aimed at determining how CAFs redirect glutamine into proline biosynthesis (Aim 1) and identifying the adaptive mechanisms by which CAFs sustain collagen production when extracellular glutamine is limited (Aim 2). Finally, I will investigate whether targeting these adaptive mechanisms is a strategy to reduce desmoplasia in PDAC without affecting matrix production in healthy tissues (Aim 3). The scientific knowledge gained from these studies as well as my research training plan will help me to develop a unique research program and facilitate transition into to independence, with the long-term goal to study the mechanistic basis of tumor-stroma crosstalk and to apply this knowledge to develop therapeutic strategies that can improve outcomes of cancer patients. In addition to my research objectives, I have outlined a detailed career development plan that will help me obtain important skills for leading an independent research laboratory, including teaching and mentoring skills, scientific communications, grant writing and laboratory management. I will work towards my goals under the mentorship of Dr. Craig Thompson, a leader in cancer metabolism with a stellar track record of trainees that went on to faculty positions. In addition, I have assembled an Advisory Committee that will collaborate and meet with me regularly to help me perform the proposed experiments and develop into an independent researcher. This team includes Dr. Justin Cross with expertise in metabolomics, Dr. Scott Lowe with expertise in mouse models of cancer, Dr. Joshua Rabinowitz with expertise in studying metabolic flux in vivo, and Dr. Gina Sizemore with expertise in studying tumor-stroma interactions. My research and career development plan, together with my mentor, advisors and the exceptional academic environment at Memorial Sloan Kettering Cancer Center will provide a solid ground on which I can build a career as an independent investigator in cancer metabolism.

NIH Research Projects · FY 2026 · 2023-01

Project Summary Asthma is a chronic inflammatory lung disease that currently affects an estimated 8.4% of children in the U.S., resulting in over 600,000 emergency department visits and over 75,000 hospitalizations each year (Moorman et al., 2012). For decades the burden of asthma in the U.S. has fallen disproportionately on Black children living in low- resourced communities, for whom the prevalence, morbidity and mortality rates far exceed those for White children. Black children have 2-3 times higher rates of hospitalization and emergency department visits compared with White children, and a nearly 5-fold increase in asthma-related mortality (Volerman, et al., 2019). In some neighborhoods on the South Side of Chicago the rates of asthma among young Black children are over 40% (Gupta et al., 2008). The high rate of asthma and allergy in Black children appears to be in part influenced by prenatal stress exposure. Thus, an intervention designed to improve prenatal stress regulation in Black women may also reduce the risk of asthma in their children. The Nutrition and Pregnancy Study (NAPS) (NCT02647723) is a nearly completed double-blind, randomized controlled study (RCT) of prenatal fatty acid supplementation aiming to improve birth outcomes and infant neurodevelopment via improved prenatal stress regulation. Docosahexaenoic acid (DHA), an omega-3 fatty acid, has been shown to protect against the development of asthma and atopy in controlled animal studies and in observational studies of humans. Results from two small RCTs of infant formula supplementation provide further support for the protective effects of dietary fatty acids against asthma (Birch et al., 2010; Foiles et al., 2016). The NAPS study provides an unprecedented opportunity to test the effects of prenatal DHA levels on childhood asthma and atopic disease, and to further examine how patterns of prenatal stress regulation impact that effect. We propose to conduct clinical assessments of asthma and other atopic outcomes in 130 children at 5-6 years of age whose mothers participated in the NAPS study, and to collect data from medical records and maternal report to test two aims. Aim 1: Test whether prenatal fatty acid levels are associated with asthma and atopic disease in early childhood. We hypothesize that higher prenatal blood levels of DHA will be associated with lower levels of asthma and atopy in the offspring as measured via laboratory assessments of asthma and allergic sensitization at ages 5-6 years, as well as medical record abstraction and maternal report. Aim 2: Test whether prenatal stress regulation impacts the association between prenatal fatty acid levels and asthma and atopic disease in early childhood. Our primary hypothesis is that cortisol and cytokine levels, heart rate variability, and perceived stress during pregnancy will partially mediate the association between prenatal DHA levels and childhood asthma and atopic disease. We will also test whether the association between prenatal DHA levels and early childhood asthma and atopic disease is moderated by prenatal stress regulation at baseline and by changes in stress regulation during pregnancy. To our knowledge, this will be the first test of prenatal fatty acid levels on asthma and allergy in Black children living in low-resourced environments in the U.S.

NIH Research Projects · FY 2026 · 2023-01

PROJECT SUMMARY Germinal centers (GCs) drive adaptive humoral immunity by selecting for B cells with high affinity antibodies and producing memory B cells and plasma cells. We previously demonstrated that GC B cells can be subdivided into three subpopulations that are functionally, spatially, transcriptionally, and epigenetically distinct. Thus, we asked whether these unique epigenetic states are important for GC B cell function and differentiation. Our lab has also previously characterized how the epigenetic reader BRWD1 regulates epigenetic states and enhancer accessibility during the transition from large pre-B cells to small pre-B cells during B cell lymphopoiesis. Because BRWD1 is highly expressed in GC B cells, we hypothesized that BRWD1 regulates chromatin accessibility in GC B cells and is important for peripheral B cell differentiation. To study this, we generated Brwd1floxed mice to delete Brwd1 in different B cell populations. First, deletion of Brwd1 in follicular B cells inhibited GC responses with fewer GC B cells and smaller GCs observed by microscopy. Second, deletion of Brwd1 in GC B cells promoted proliferation of GC B cells and blocked differentiation of Brwd1-/- GC B cells into memory B cells without disrupting GC architecture. Furthermore, loss of Brwd1 caused an epigenetic collapse whereby differential chromatin accessibility between GC B cell subpopulations was lost. From this preliminary data, we propose a model where BRWD1 regulates chromatin accessibility at enhancers and transcription factor binding motifs in a manner critical for the cell fate decisions of peripheral B cells. To test this model, first we will further study how BRWD1 is important for B cell function and differentiation. In follicular B cells, we will characterize whether BRWD1 is necessary for either the initial differentiation or early expansion and proliferation of GC B cells (Aim 1). In GC B cells, we will study whether BRWD1 is important for somatic hypermutation and affinity maturation (Aim 2A). Furthermore, we will determine whether BRWD1 is necessary for development of pre-memory B cells or memory B cell subsets (Aim 2B). Finally, we will characterize how BRWD1 establishes epigenetic states between GC B cell subpopulations (Aim 3). We will characterize how BRWD1 binds at different sites and how the histone marks that recruit BRWD1 change between GC zones. We will identify active and repressed enhancers alongside our chromatin accessibility data to study how BRWD1 coordinates changes in enhancer accessibility between GC B cell subpopulations. Completion of this proposed project will provide insight into fundamental mechanisms central to the humoral immune response, autoimmunity, and lymphoma development within the GC.

NIH Research Projects · FY 2025 · 2022-12

PROJECT SUMMARY/ABSTRACT RNA processing is an essential cellular process that when dysregulated underlies the development of neurological diseases. Several mutations in nuclease containing complexes cause pontocerebellar hypoplasia (PCH), a severe neurological disorder that often leads to prenatal death. Most cases of PCH are linked to mutations in the tRNA Splicing Endonuclease (TSEN) Complex, which is responsible for the cleavage of tRNA introns prior to tRNA maturation, and its accessory protein, CLP1, which is a critical negative regulator of tRNA splicing. Genetic deletion of any single TSEN protein in yeast, engineered to have tRNAs without introns, was shown to be lethal, suggesting that the TSEN complex likely has substrates beyond the tRNAs, which may underlie the development of PCH. Likewise, mutations in another nuclease, target of Egr 1 (TOE1), a deadenylase and 3’-exonuclase, are also are linked to PCH. TOE1 is the only deadenylase believed to mature snRNAs, but it also moonlights in other cellular pathways, highlighting how much is yet to understand about its role in protein complexes. To determine how mutations in TOE1, CLP1, and TSEN proteins lead to PCH, there remains a critical need to understand how these complexes assemble, recognize and process RNAs, and how their enzymatic activity is regulated. Characterizing healthy cellular roles of these proteins is essential to determining how their dysfunction causes PCH. We aim to address these critical questions through the following proposed Aims. In Aim 1, Structural and molecular techniques will be used to identify how the TSEN complex recognizes and processes tRNAs and other RNA substrates. In Aim 2, we will determine how the CLP1/TSEN complex are regulated at the molecular and cellular level and how PCH mutations disrupt their regulation. Further, in Aim 3, we will identify how PCH mutations alter TOE1 function and regulation, using proteomics and molecular biology approaches. The proposed work is significant because it will provide a structural description for how known PCH mutations may interfere with complex formation, stability, or function for a range of PCH-linked proteins. This work will further provide insight into shared mechanisms by which these protein complexes cause PCH. Furthermore, the work here will characterize new RNA processing roles for these nucleases.

NIH Research Projects · FY 2026 · 2022-12

Project Summary/Abstract: Despite extensive studies on relationships between diets and cancer risk, or many “balanced” nutrition therapies with hope to keep cancer patients healthy and strong for treatment and recovery, little is known about how dietary substances influence cancer. Our recent work supports a novel concept that acetoacetate, a diet-derived, circulating ketone body, and chondroitin sulfate, a dietary supplement, function as signaling molecules and selectively promote BRAF V600E-expressing tumor growth. This lays the foundation for our central question that is: which circulating diet-derived substances - defined as “blood chemicals”, commonly containing diet- derived nutrients including inorganics, organic metabolites, lipids, dietary supplements and proteins - potentiate or attenuate cancer initiation, progression or responses to anti-cancer therapies, and how? We thus constructed a “blood chemical (BC)” compound library and performed two preliminary screens to identify BCs that influence responses to immune checkpoint inhibitors (ICIs). We identified trans-vaccenic acid (TVA; a.k.a. (11E)-octadec- 11-enoic acid) as an “overlapping” top candidate from both screens, which not only enhances activation of T cells but also “rescues” PD-L1/PD-1-dependent exhaustion of T cells. TVA is the predominant form of trans- fatty acids enriched in human milk, while cis-vaccenic acid (CVA), a stereoisomer of TVA, is found in Sea Buckthorn oil. TVA is also commonly found in dairy products including milk and butter. TVA is relatively stable, and naturally only ~19% or 12% of dietary TVA is converted to rumenic acid in human or mice, respectively. Using diverse immunogenic and immunodeficient mouse models, we found that TVA, but not CVA, enhances anti-tumor immunity via CD8+ T cells. Mechanistically, TVA exhibits extracellular signaling function and enhances CD8+ T cell activation through a G-protein-coupled receptor (GPCR)-cAMP-responsive element binding protein (CREB) pathway. Moreover, we identified immunosuppressive GPR43, a short chain fatty acid (SCFA)-binding GPCR, as a target of TVA. Taken together, we hypothesize that dietary TVA functions as a signaling molecule to potentiate activation of CD8+ T cells by attenuating GPR43, leading to enhanced anti-tumor immunity. Thus, TVA’s effects on T cells are independent of the PD-L1/PD1 axis, providing a perfect rationale to evaluate potentially synergistic efficacy of TVA in combination with immune checkpoint therapy for an improved immunotherapy. Three specific aims include: (1) To test the hypothesis that dietary TVA enhances CD8+ T cell activity and consequent anti-tumor immunity as a single agent, and has synergistic effects in combination with ICIs; (2) To test the hypothesis that dietary TVA exhibits extracellular signaling function through a GPCR-CREB axis for CD8+ T cell activation, and explore the underlying signaling and epigenetic mechanisms by temporal, integrated mechanistic studies; and (3) To test the hypothesis that TVA attenuates GPR43 by competing with its SCFA agonists, and perform structure-activity research (SAR) to design TVA-derivatives with improved efficacy to target GPR43 and consequently activate CD8+ T cells.

NIH Research Projects · FY 2026 · 2022-12

Project Summary Adipose tissue is a central player in energy balance and glucose homeostasis, it is able to expand in the face of caloric overload in order to store energy safely, but it can become overloaded and dysfunctional, leading to systemic metabolic compromise in the form of insulin resistance and Type 2 diabetes. To investigate differences in individual cell types in lean and obese individuals, I have performed single nucleus RNA sequencing (sNuc- seq) on human subcutaneous and visceral white adipose tissue and created an atlas of cell types present in white adipose tissue. A major finding from this work was the identification of distinct subpopulations of adipocytes, some of which are associated with body mass index (BMI). By associating our data with genome- wide association studies (GWAS) for metabolic traits such as T2D, we additionally predict that some adipocyte subpopulations are associated with metabolic disease. The objective of this project is to identify factors that predispose one subpopulation over another, both externally as well as transcriptionally. To do this I will perform sNuc-seq on adipose tissue collected from subjects during and post-bariatric surgery in order to characterize the change in adipocyte subpopulation during weight loss. I will next interrogate the chromatin state of adipocyte subpoulations by performing the Assay for Transposase Accessible Chromatin on single nuclei from adipose tissue of lean and obese individuals. Finally, I will directly test potential signaling and transcriptional regulators of subpopulation identity by performing a screen of potential signaling regulators as well as a CRISPRa screen of potential transcriptional regulators to try to recapitulate distinct subpopulations in vitro. The experience that I have in characterizing adipose tissue at single cell resolution, as well as the experience of my mentor and co- mentor in studying transcriptional and genetic regulation of adipocytes make me uniquely positioned to answer these questions. Taken together, these studies will enhance our knowledge of human white adipocyte diversity and will set the stage for downstream studies in my own independent lab and in the community at large.

NIH Research Projects · FY 2026 · 2022-12

PROJECT SUMMARY A longstanding question in developmental biology is how stem cells commit to a certain lineage. While cell surface markers correlate with hematopoietic stem cell (HSC) functions, less is known regarding transcriptional regulators that drive stem cell behavior. CUX1 encodes a highly conserved homeodomain-containing transcription factor. Deletions and inactivating mutations of CUX1 are recurrent in clonal hematopoiesis of indeterminate potential and myelodysplastic syndrome, and CUX1 loss is associated with cytopenias, lineage skewing, and a poor-prognosis in disease states. Typically, only one copy of CUX1 is inactivated, indicating that loss of CUX1 impacts hematopoietic development in a haploinsufficient manner. In line with this, we previously reported a dosage-dependent role for CUX1 in hematopoiesis. How CUX1 levels regulate hematopoietic stem and progenitor cell (HSPC) fate, with respect to proliferation, self-renewal, and lineage choice remains a major gap in knowledge. The overall objective of the current proposal is to determine the molecular mechanism by which CUX1 dosage regulates HSPC fate. To address this, we have now generated a novel CUX1-reporter mouse to measure CUX1 protein levels at the single-cell level during development. In preliminary data, we demonstrate that CUX1 is expressed highly in HSPCs while exhibiting heterogeneity within stem and progenitor compartments. We show that CUX1 levels correlate with hematopoietic stem cell (HSC) activity, with CUX1dim HSCs demonstrating higher multilineage, long-term hematopoiesis compared to CUX1bright HSCs. We show that CUX1 interacts with the SWI/SNF chromatin remodeling complex to regulate chromatin accessibility in primary human HSPCs. Thus, our central hypothesis is that CUX1 regulates HSC fate via dosage-dependent opening of lineage-specific enhancers through recruitment of SWI/SNF. We test this hypothesis with two Specific Aims. Aim 1: Overall hypothesis – CUX1 upregulation is necessary for HSC differentiation. In a series of in vivo experiments in mice and complementary assays with primary human cells, we will correlate CUX1 protein levels with HSC functions, including proliferation, self-renewal, and differentiation. We will perturb CUX1 levels to assess how CUX1 dosage impacts these functions. Aim 2: Overall hypothesis – CUX1 upregulation drives enhancer priming for differentiation. To determine how CUX1 impacts chromatin remodeling and enhancer activation, we will leverage cutting-edge functional genomics approaches and single-cell methodologies in HSCs using our CUX1-reporter and CUX1-knockdown mice and primary human HSPCs. Accomplishing the proposed studies will elucidate the critical epigenetic role for dosage-sensitive CUX1 transcriptional regulation in hematopoietic development. This work is critical to achieve our long-term goal of identifying therapeutic interventions for patients with CUX1 haploinsufficiency.

NIH Research Projects · FY 2025 · 2022-11

Project Summary Alterations in protein-protein interactions (PPI) can result in dysregulated biological signaling. PPIs are critical drivers of a plethora of disease states and are increasingly recognized as important therapeutic targets. However, PPIs have been difficult to target with traditional therapeutics, and are often considered “undruggable”, because traditional small molecule discovery strategies are not well suited to identify molecules with suitable properties to disrupt PPIs. When a disease-associated PPI target is identified, translating that information into a validated molecular probe can take years, development of a clinically useful therapeutic can take decades, and even worse, in most cases these efforts fail entirely. The costs associated with developing therapeutic leads limits pursuits towards only thoroughly validated targets. A faster, less expensive, and more efficacious pipeline for PPI inhibitor discovery would allow researchers to quickly identify probes for directly perturbing PPIs in relevant model systems to assess the validity of the PPI as a therapeutic target and to generate candidate inhibitors for clinical development. While almost all areas of basic and translational biology research have benefitted from 21st century technological advances in genetic sequencing, mass spectrometry, and other diagnostics, drug discovery still largely relies on 20th century methods, which have proven to be frustratingly slow and unsuccessful in critical ways—our proposed technology aims to solve these problems. We hypothesize that continuous evolution techniques will allow us to rapidly evolve PPI inhibitors for a wide range of cytosolic protein targets. Specifically, we will pursue two key advancements to realize this broad goal. In Aim 1, we will demonstrate that non-continuous selection followed by phage-assisted continuous evolution (PACE) allows us to rapidly evolve protein binders for diverse proteins using a library-of-libraries approach. This approach will eliminate a crucial bottleneck in PACE, optimization of initial selection stringency, and bring PACE much closer to automated plug-and-play allowing for broader adoption of this technique. In Aim 2, we will construct and validate a PACE compatible biosensor linking disruption of a specific PPI to phage fitness allowing for us to directly select for PPI inhibitor function. Our goal is to demonstrate generation and validation of high potency PPI inhibitors for a given target in under 1 month. We will validate these evolution platforms using three well-studied oncogenic protein-protein interactions that have been the focus of small molecule drug development for decades: MDM2-p53, KRAS-RAF, and Myc-MAX. While this is a lofty goal, the power of evolution has long been recognized as a promising solution to this problem; we are hopeful that by merging innovative biosensor designs and continuous evolution, we can unlock the full potential of laboratory evolution. If successful, these platforms have the potential to revolutionize the drug discovery paradigm and accelerate the discovery of novel PPI inhibitors.

NIH Research Projects · FY 2025 · 2022-09

Project Summary/Abstract Hematopoietic stem cells (HSCs) exhibit heterogeneity with respect to repopulating capacity, lineage bias, and cell cycle participation. Teleologically, heterogeneity is the result of stem cells making fundamental decisions regarding the population of the tissue. Differences in stem cell behavior have been linked to the expression of cell surface and cytoplasmic markers without mechanistic explanation. HSC heterogeneity has not previously been attributed to a transcription factor. We propose the observed heterogeneity in HSCs can be explained by the actions of a dose-dependent, homeodomain-containing transcription factor, CUX1. Three lines of evidence suggest CUX1 to be a putative orchestrator of HSC heterogeneity: it recurrently acts in a dose-dependent manner across developmental systems, its recurrent loss in high-risk hematopoietic disease, and its role in chromatin regulation. I report the generation of a CUX1mCherry reporter mouse to study the role of CUX1 in hematopoiesis. The Cux1mCherry mouse is the result of an in-frame, C-terminal mCherry tag at the endogenous CUX1 locus. The addition of the tag creates no aberrant hematopoietic phenotype, suggesting this is a suitable model for the proposed studies. The immunophenotypic long-term HSC (LT-HSC) compartment has among the greatest variances in CUX1 expression. Across CUX1 protein levels in the LT-HSC compartment, we report several correlations to strongly suggest that the observed variation in CUX1 protein results in meaningful differences in stem cell behavior. For example, CUX1Bright HSCs are more likely to be cycling than CUX1Int and CUX1Dim HSCs at steady state. Thus, our studies suggest that CUX1 is playing a dose-dependent role in murine hematopoietic stem and progenitor cells (HSPCs). This proposal aims to (i) establish the role of CUX1 in lineage bias and repopulating capacity and (ii) determine the mechanism by which CUX1 exerts a dose- dependent role. Accomplishing the proposed studies will illuminate an important paradigm in developmental biology: how a small pool of stem cells balance self-renewal and differentiation to give rise to all the mature cells in a tissue. An etiological understanding of stem cell behavior will provide new insights into the development of new therapies for the many diseases that arise in HSCs. The project I propose here is accompanied by a training plan developed by me and my mentors that delineates four goals I will need to accomplish to propel me towards becoming a successful independent physician-scientist. These four goals include gaining expertise in hematopoiesis, gaining expertise in bioinformatics, developing proficiency in scientific communication, and integrating the scientific and clinical aspects of my training. Realizing these goals will give me the skills that I need to be a physician-scientist well- equipped to address meaningful biological questions related to the catastrophic illnesses of childhood.

NIH Research Projects · FY 2024 · 2022-09

ABSTRACT Maternal mortality and severe maternal morbidity (SMM) continue to rise in the United States, and women from low-income communities, minoritized racial/ethnic groups, and those with Medicaid or who are uninsured face increased risk. Hospital care at the time of delivery, and prenatal care during pregnancy, both occur late in the progression of conditions that are important to maternal outcomes. Therefore, expanding access to preconception care is a promising approach to improving maternal health and outcomes. Evidence-based preconception interventions include counseling patients on healthy behaviors (e.g. taking folic acid), managing chronic health conditions, and providing contraceptive care to plan timing of pregnancy. For many low-income women, a major barrier to receiving preconception care has been lack of health insurance coverage. The Affordable Care Act (ACA) reduced the uninsured rate among reproductive age women, giving women greater access to preconception care. Some states began implementing the ACA’s Medicaid coverage expansion in 2014, and women’s report of receiving counseling on preconception health increased significantly in Medicaid expansion states compared to non-expansion states. Prior research has identified an association between preconception care and decreased risk of SMM, particularly among women with a chronic disease. However, further research is needed to establish a causal effect. This study will harness the study team’s access to nationwide Medicaid claims data and leverage Medicaid expansion as a natural experiment to estimate the causal effect of preconception care on SMM for women insured by Medicaid. Analyzing existing Medicaid claims data will allow us to describe change in preconception care and SMM rates in expansion and non- expansion states (Aim 1). We will then use quasi-experimental design enabled by variation in state policy (i.e. timing of adopting Medicaid expansion) to estimate the causal effect of preconception care on risk of SMM, both in the Medicaid population overall and in sub-groups at highest risk based on medical history (e.g. chronic disease or history of prior SMM) or sociodemographic characteristics (Aim 2). We will also gain the nuanced perspectives of patients at high risk for SMM and clinicians who care for them, using qualitative focus groups and key informant interviews (Aim 3). By combining quantitative Medicaid claims and qualitative analyses, we will identify preconception interventions and approaches with high likelihood of reducing SMM, particularly for women at highest risk. We will translate these findings to develop a novel preconception care model that will be tested in future work.

NIH Research Projects · FY 2025 · 2022-09

Over the past two decades, rural counties in the U.S. have seen increasing rates of overdose death along with rising hepatitis C incidence and outbreaks of HIV linked to injection drug use. The rural opioid crisis is co- occurring and intertwined with evolving methamphetamine use and polysubstance use more broadly. The burden of morbidity and mortality related to substance use in the rural setting is exacerbated by the scarcity of resources such as addiction treatment, overdose education and naloxone distribution, and other prevention services. Given the lack of biomedical treatment for methamphetamine use disorder, efforts to improve health outcomes for people who use methamphetamine hinge largely on such strategies. In the face of the pandemic grassroots HRS organizations have been challenged to adapt service delivery amidst sparse resources and shifting drug use behavior. The proposed study aims to understand evolving drug use and prevention behavior in rural settings, including those where polysubstance use with methamphetamine and fentanyl are prevalent, in order to inform the expansion of effective services within these communities. We will engage community providers and participants in a process of tailoring strategies for remote, contactless delivery of services, and evaluate their implementation. This study will explore individual, interpersonal, and community influences on engagement in prevention behavior according to the socio-ecological model. In Aim 1, we will assess drug use, risk behavior, and decision-making processes regarding prevention service engagement and use and distribution of supplies among people who use drugs (PWUD). We will collect data through multiple methods including baseline surveys, two weeks of daily assessments using mobile phones, and semi-structured interviews with PWUD. In Aim 2, using social network survey methods as well as in depth qualitative interviews, we seek to understand the dynamics of secondary distribution by identifying the personal and social network characteristics of participant champions who may implement network interventions. In Aim 3, we will develop and evaluate the implementation of remote service delivery including digital lock boxes tailored to address structural barriers in rural settings. This process will include needs assessment through key informant interviews, guided discussion groups forming a learning community co-led by a community provider, and implementation and evaluation informed by the EPIS implementation framework. This work will provide foundational knowledge to develop practical service delivery strategies for rural areas facing disparate challenges in pandemic-era service provision as well as inform future network interventions for rural drug use characterized by methamphetamine and the ubiquitous presence of synthetic opioids.