Washington University

NIH Research Projects · FY 2025 · 2018-05

ABSTRACT/SUMMARY The Washington University “Multidisciplinary Training in Alzheimer and Related Dementias” (ADRD-T32) Training Plan is designed to train the next generation of translational researchers in Alzheimer disease and related dementias (ADRD). The program will be led jointly by the Knight ADRC and the Hope Center for Neurological Disorders at Washington University School of Medicine (WUSM). The focus of the program is on multidisciplinary team-based research that accelerates the development of treatments for ADRD and increases diversity in the research workforce by enhancing the clinical and translational basic science experience for young investigators in a highly enriched research environment. The Knight ADRC and the Hope Center support a highly collaborative environment that exemplifies multi-disciplinary team-based research. Thirty-three faculty from twelve Departments at Washington University will mentor trainees in dementia research. The faculty include basic scientists, translational neuroscientists and clinical investigators. WUSM has outstanding resources, including state of the art facilities for translational research, biostatistics and informatics, and an excellent track record in developing new therapies. The predoctoral students will be drawn from 9 different doctoral programs, including those affiliated with specific departments (Biomedical Engineering, Psychological and Brain Sciences, and Social Work) as well as from programs in the Division of Biology and Biomedical Sciences, including Biochemistry, Computational and Systems Biology, Developmental, Regenerative and Stem Cell Biology, Human and Statistical Genetics, Molecular Genetics & Genomics, Neuroscience and from the McKelvey School of Engineering. Postdoctoral trainees will be drawn from Chemistry and Developmental Biology, clinical departments (Neurology, Medicine, Pathology, Psychiatry and Radiology), Biostatistics, Psychological and Brain Sciences and the School of Social Work. The training program will support 3 predoctoral trainees and 3 postdoctoral trainees; each year the program will support 2 medical students from Meharry Medical College School of Medicine for a short-term training program aimed at exposing promising medical students to the field of ADRD early in their medical training. The program will provide training (but no stipend) for two Washington University School of Medicine medical students participating in the Year Long Research Program. The training program includes sessions devoted to clinical aspects of dementia, datasets and resources available at the Knight ADRC, WUSM and through large national efforts, bioinformatics, and responsible conduct of research. The program aims to develop a pipeline of diverse and rigorously trained ADRD researchers through mentorship with leading experts in all facets of ADRD research from a leading center with over 4 decades of success in training AD researchers.

NIH Research Projects · FY 2026 · 2018-05

Project Summary/Abstract Sepsis is defined as a dysregulated host inflammatory response that occurs due to life-threatening infection with the presence of organ dysfunction. Sepsis is the most frequent cause of mortality in most intensive care units and is responsible for over a quarter million deaths in the United States annually. The incidence of sepsis is increasing because of the aging population and the associated weakening of the immune system that occurs in the aged. Until recently, most research on sepsis was focused on blocking the initial hyper-inflammatory cytokine-mediated phase of the disorder. Improved treatment protocols have resulted in most patients surviving this initial hyper-inflammatory phase of sepsis and entering a protracted immunosuppressive phase. The majority of deaths in sepsis occur during this immunosuppressive phase of the disorder. Deaths in this immunosuppressive phase of sepsis are typically due to failure to control the primary infection or a result of acquisition of secondary hospital-acquired infections, often with opportunistic pathogens thereby underscoring the host’s impaired immunity. The reactivation of multiple latent viruses including cytomegalovirus and herpes simplex virus that occurs in patients with protracted sepsis further attests to the profound degree of immunosuppression in these patients. There is a growing body of evidence in animal studies as well as data from small phase II clinical trials indicating that therapies which boost the host immune system can improve morbidity and mortality in sepsis. This immuno-therapeutic based approach to sepsis has been the focus of the principal investigator for over two decades. The current proposal is an extension of these investigations. The overarching goal of this proposal is to identify molecular mechanisms of immunosuppression in sepsis and develop new immuno-adjuvant therapies that restore host immunity, ameliorate organ injury, and improve survival in sepsis. We are focusing on testing immune modulatory agents that have an excellent safety profile and are in current clinical trials. Successful completion of this study would enable rapid translation of newly identified drugs into clinical trials in sepsis and offer a new way forward against this heretofore intractable disease.

NIH Research Projects · FY 2025 · 2018-05

Project Summary The environment is a primary determinant of bacterial growth and behavior. It impacts not only metabolism and growth rate, but also morphology, development, and antibiotic susceptibility. Our research centers on the essential processes that coordinate cell growth and cell cycle progression with the environment. In bacteria these processes are critical, maximizing proliferative potential across a wide physicochemical landscape. Defects in these processes can be catastrophic, impairing growth, reducing viability, and rendering cells hypersensitive to antimicrobials. The increasing prevalence of antibiotic resistant pathogens provides a powerful motivation for our work. Understanding how bacteria adapt to the physicochemical environment should reveal novel approaches for the treatment of bacterial pathogens and identify targets for the development of next-generation therapeutics. In this this effort we employ evolutionarily distinct organisms including two Gram-negative Gammaproteobacteria: the model organism Escherichia coli and the pathogen Klebsiella pneumoniae. In 2019 K. pneumoniae was the third leading cause of global deaths due to or associated with antibiotic resistance (Lancet, 2022). The cell envelope is vital to bacterial viability, required as both a permeability barrier and a buttress against turgor pressure. Exposed to the environment, essential cell envelope processes must remain functional across a large range of conditions. Understanding the factors that underlie and preserve cell envelope integrity is a primary research focus. Extending our finding that functional redundancy amongst E. coli cell wall enzymes ensures robust growth across pH values, we will use single molecule techniques and biochemistry to illuminate the impact of pH on the activity of individual PBP transpeptidases in situ. These enzymes play a critical role in peptidoglycan cross linking and are the target of beta-lactam antibiotics. We will also identify factors underlying pH-mediated changes in cell wall synthase activity and beta-lactam resistance in K. pneumoniae. As part of a broader effort to understand the mechanisms coordinating cell envelope biogenesis with cell growth and cell cycle progression, we will elucidate the essential role of the outer membrane component lipopolysaccharide (LPS) in E. coli biology. Finally, building on our longstanding interest in the relationship between nutrient availability and cell morphology, we will characterize the mechanisms governing a newly identified starvation-mediated stress response, cytoplasmic condensation, and identify the effector(s) responsible for the positive relationship between the starvation-associated alarmone, ppGpp, and cell division. We are aided in these endeavors by our multidisciplinary approach that employs a diverse array of techniques, as well as an extensive network of close colleagues and collaborators with whom we enjoy a free exchange of reagents and ideas. These advantages allow us to answer previously intractable questions in physiology and homeostatic control that are relevant to organisms throughout the tree of life.

NIH Research Projects · FY 2026 · 2018-04

PROJECT SUMMARY/ABSTRACT Despite increased funding across the NIH enterprise over the past decade, fewer physician trainees are choosing to pursue scientific careers, contributing to a decline in the number of physician-scientists capable of achieving independent research success. To address the workforce needs, the Department of Psychiatry at Washington University launched the Psychiatry Resident Research Education Program (PRREP) in 2016. Funded by NIMH’s R25 program in 2017, PRREP is agnostic of prior research experience, is available to both our general and child and adolescent psychiatry (CAP) training programs, and provides individualized educational planning divided into two phases. Phase 1 aims to increase the number of incoming residents considering a research career by enhancing recruitment and providing early education (career counseling/mentoring, didactics, seminars, on-line coursework) and introductory research experiences. The more research-intensive Phase 2 focuses on gaining experience in key areas, including study design, regulatory considerations, budgeting, data collection/management, and analysis. Total time spent in hands-on research will be approximately 12 months for a resident in the 4-year general adult program and 13 months for a CAP resident who is in training for 5 years. During this first period of funding, we have doubled the number of graduates pursuing research careers (50% with a Ph.D.). Our PRREP trainees have been listed as an author on manuscripts 101 times. In this renewal application we seek on-going support from NIMH’s R25 program to expand on the early successes we have achieved with this critical program, building upon the solid foundation we have established in these first 4 years. In this renewal application, we propose to continue to recruit promising trainees into PRREP, providing early and focused mentorship and hands-on research experiences to promote development of skills and knowledge necessary for successful transition to Phase 2. To build upon the success achieved during the initial funding period, we will focus on the following goals: 1) we will target recruitment of individuals interested in CAP research careers, provide early exposure to CAP faculty and enhance the mentorship of trainees interested in CAP; 2) utilizing institutional training and support resources, we will promote excellence in mentorship by providing mentors feedback throughout the academic year, and provide mentorship training and support for early-career faculty with emerging mentoring skills; and 3) informed by the initial 4 years of funding, we will develop, implement and track educational milestones targeting research skills needed to become a successful principal investigator. Funding for the next 5 years will provide us with the means to build on this solid foundation to continue providing innovative and inclusive research training for the next generation of physician-scientists in psychiatry.

NIH Research Projects · FY 2026 · 2018-02

The annual Society for Reproductive Investigation (SRI) meeting brings together both clinical and basic scientists from around the world to discuss research related to women's health and reproductive science. The highly successful annual SRI meeting has had an average attendance of 1062 investigators over the last five years and has brought together established senior and junior investigators to report and discuss their findings in an atmosphere conducive to frank yet amicable exchange. This application seeks funding to cover travel costs to allow seven trainees and new investigators to attend SRI annual meetings. This proposal also seeks funding to support the travel costs of a senior US-based investigator who will present an invited Distinguished Presidential lecture at the meeting. Meetings are scheduled in March of each year in Brisbane, Australia (2023) and Vancouver, British Columbia, Canada (2024). The four-day meeting includes two or three Distinguished Presidential Lectures, oral and poster presentations, 12 mini- symposia, a new investigator plenary, career development forums, and networking events.

NIH Research Projects · FY 2025 · 2018-02

PROJECT SUMMARY Effective treatment remains an unmet and urgent need for patients with pancreatic ductal adenocarcinoma (PDAC). Unlike most cancer types where targeted therapies and immunotherapy have become standard treatments, PDAC is unique among major cancers in still relying almost entirely on chemotherapy for disease control. The administration of multiple cytotoxic agents, while necessary for efficacy, results in significant clinical toxicities, and treatment response rates remain low at 30–40%, with responses often lacking durability. Therefore, achieving therapeutic breakthroughs necessitates novel biological discoveries in pancreatic cancer. Over the past decade, our lab has generated substantial preclinical and clinical evidence showing that innate immune signaling pathways in PDAC cells are critical in establishing an immunosuppressive tumor microenvironment that inhibits anti-tumor T cell activity. We initially focused on IRAK4, a kinase regulating Toll-like receptor and IL-1 receptor signaling and demonstrated that targeting IRAK4 enhances chemotherapy efficacy and may potentiate immunotherapy. This work laid the groundwork for three clinical trials. Building on these insights, we have identified additional signaling nodes related to IRAK4, with MAP3K8 (TPL2) emerging as a particularly promising target. Our preliminary data indicate that TPL2 regulates multiple signaling pathways in PDAC cells, including the processing of micro(mi)-RNA species that may endow these cells with aggressive and immunosuppressive phenotypes. We found that targeting TPL2 enhances the efficacy of FOLFIRINOX chemotherapy and reprograms the tumor microenvironment, potentially enabling checkpoint immunotherapy to be effective in PDAC. Based on these findings, our team proposes three Aims: Aim 1: We will investigate the mechanism by which TPL2 controls miRNA processing, particularly through modulation of AGO2 and AGO3 function. This study will provide one of the few comprehensive analyses of miRNA's role in PDAC biology. To ensure success, we will collaborate with an experienced RNA biologist, Dr. Sergej Djunarovic. Aim 2: We will explore the role of TPL2 in PDAC cells, as well as in myeloid and T cells, using a conditional TPL2 knockout mouse strain provided by Dr. Andrew Greenberg. These experiments will elucidate how TPL2 influences PDAC tumor progression and inform immunotherapeutic strategies tested in Aim 3. Aim 3: We will evaluate the preclinical efficacy of the TPL2 inhibitor tilpisertib combined with FOLFIRINOX in 20 patient-derived xenograft models. We will also identify a rational immunotherapeutic combination to be tested in the immunocompetent genetic (KPPC) mouse model.

NIH Research Projects · FY 2026 · 2018-01

In the decades ahead, demographic trends suggest that the sheer number of older adults will increase, and they will make up a larger proportion of the overall population. Many of these individuals will experience some type of neurological disease at some point in their life. Consequently, we need an expanded cadre of scientists trained to generate new knowledge about neurological conditions and aging. To address this need, we will conduct an eight-week Summer Undergraduate Research Experience in Aging and Neurologic Diseases at the Harvey A. Friedman Center for Aging at Washington University. The program will 1) provide undergraduate students with a closely mentored research laboratory experience, combined with didactics and professional development sessions, and 2) frame the research training in the context of aging and neurological disorders common in later life. These goals address the need to nurture new researchers to address the workforce shortage of scientists investigating issues of an aging society. Up to 10 undergraduate students each summer will be matched with an individual faculty member whose research focuses on one of three neurological conditions (stroke, Parkinson’s disease, or dementia) or who conducts research on interventions for individuals with these conditions (e.g., home modifications, engagement in valued activities). Students will also gather for weekly seminars that expose them to a multidisciplinary perspective on aging, teach them research skills, and encourage their professional growth and momentum toward further scholarship and training.

NIH Research Projects · FY 2026 · 2017-12

Project Summary/Abstract Myeloproliferative neoplasms (MPNs) are chronic blood disorders that that can cause severe symptoms and early death. New treatments have become available that help ameliorate symptoms, but they do not reliably slow or halt disease progression. We seek to better understand what drives disease development and progression in MPNs, so that we can develop better therapies for patients with these diseases. Our preliminary data indicates that the NFkB pathway is abnormally activated in MPNs. We hypothesize that NFkB pathway signaling contributes to the development of MPNs. Therefore, we have proposed a combination of mouse and human studies to determine how NFkB contributes to MPN pathogenesis, and to evaluate whether inhibition of NFkB pathway signaling may have potential therapeutic benefits for MPN patients.

NIH Research Projects · FY 2025 · 2017-12

PROJECT SUMMARY/ABSTRACT Background − Maintenance of proteostasis is central to cellular fitness and is achieved through sophisticated protein quality control (PQC) pathways that remove dysfunctional and unwanted proteins and protein complexes that become cytotoxic if allowed to accumulate and condense. In fact, protein aggregation encouraged by PQC defects is a hallmark of aging, cancer, and numerous human ‘aggregation- prone’ pathologies, including amyotrophic lateral sclerosis, Alzheimer’s, Parkinson’s and Huntington’s diseases, and related multisystem proteinopathies. Consequently, full understandings of PQC could offer new strategies to mitigate protein aggregation and subsequent proteotoxic stress. Previous Work − We discovered mechanistically conserved PQC routes that direct the autophagic elimination of inactive proteasomes and the CDC48 segregase (p97/VCP in humans), which offer experimentally robust models for describing defective protein clearance. Notably, turnover of both protein complexes shares features with amyloidogenic protein removal, including sequestration, ubiquitylation, and subsequent recognition by dedicated autophagic receptors, which for proteasomes also requires a trio of ubiquitin ligases that likely work in concert to assemble appropriate poly-Ub chain topologies. Project Aims − This project proposes to describe in detail the autophagic clearance of proteasomes and CDC48 in both yeast and Arabidopsis, with the goal of discovering aspects central to autophagic PQC. For proteasomes, we will: (i) examine, using genetics, fluorescence microscopy, interaction studies and ubiquitin linkage mapping, where, when, and how the ligases San1, Rsp5 and Hul5 coordinately contribute to dysfunctional proteasome ubiquitylation; (ii) identify ubiquitylation linkages needed to generate autophagy competent substrates; and (iii) deduce how Hsp42-mediated sequestration into cytoplasmic membrane-less aggresomes, versus condensation into proteasome storage granules, contributes to the process. Likewise, studies on CDC48 turnover will confirm that ubiquitylation is a key signal, followed by the identification of relevant ubiquitin ligases and understanding of how CDC48 sequestration contributes to its turnover. Moreover, we will test our hypothesis that the autophagic routes used to clear dysfunctional proteasomes and CDC48 also eliminate amyloidogenic proteins that are at the heart of numerous aggregation-prone pathologies. Finally, we will further define and expand upon a new class of autophagic receptors/adaptors that use a novel interface to dock with ATG8 (LC3 in humans) lining autophagic vesicles, thus helping to increase the known reach of selective autophagy. Outcomes − Through this cumulative research, we hope to define autophagic routes relevant to aggregation-associated PQC, which will shed light on the roles of ubiquitylation, biomolecular condensation, and autophagy in mitigating proteotoxic stress and ultimately inform upon new therapeutic strategies for various amyloidogenic pathologies.

NIH Research Projects · FY 2026 · 2017-09

A. Project Summary/Abstract The goal of this renewal R01 application is to identify an 18F-labeled radiotracer to quantitatively measure sphingosine-1-phosphate receptor 1 (S1PR1) expression in multiple sclerosis (MS) patients. MS is associated with a lymphocyte-mediated autoimmune response that ultimately leads to repeating cycles of demyelination and repair. S1PR1 is extensively expressed on lymphocytes and endothelial cells and participates in this autoimmune inflammatory process by regulating immune cell trafficking in the brain. Currently, four S1P modulators fingolimod, siponimod, ozanimod, and ponesimod have been approved by FDA for treating MS, but the pathophysiologic mechanism of S1PR1 in MS is still not well understood. Positron emission tomography (PET) investigating S1PR1 function in MS and other diseases is limited by the lack of a clinically suitable radiotracer. To address this urgent clinical need, our team successfully transferred [11C]CS1P1, a C-11 S1PR1 radiotracer into clinical investigation. Our human study data confirm that liver is the critical organ for radiation dose absorption and that [11C]CS1P1 is safe for patient who receives a dose of up to 740 MBq (20 mCi). Our proof of concept brain studies showed higher uptake of [11C]CS1P1 in gray matter than in white matter, consistent with S1PR1 mRNA expression in the normal human brain. In MS patients, we observed increased [11C]CS1P1 uptake within both brain lesions and normal appearing white matter (WM). These results are consistent with extant evidence of inflammatory infiltrate in both the brain lesions and normal appearing WM in MS. Together, our data suggest that PET measurement of S1PR1 protein expression could provide a unique method to detect neuroinflammation and response in MS patients. Our initial radiometabolite analysis of human plasma discovered a lipophilic radiometabolite post injection of [11C]CS1P1 and led to initial concerns. However, we recently confirmed the molecular structure of the radiometabolite, and radiometabolite analysis of the rat brain and plasma post-injection of the radiotracer and PET brain studies of the F-18 labeled radiometaobolite demonstrated the radiometabolite does not enter into the brain. Additionally, our team has synthesized ~120 new structurally diverse S1PR1 ligands, and ~30 of them are highly potent (IC50 < 20 nM) and selective for S1PR1. To date, our preliminary data indicate that three F-18 radiotracers are promise for S1PR1 imaging, two of them are F-18 labeled using different procedures and are anticipated to have different pathway compared to [11C]CS1P1. Therefore, in this renewal phase, we will leverage our recent experience with [11C]CS1P1 and develop and transfer an 18F-labeled S1PR1 tracer for clinical investigation. We propose two specific aims: 1) Identify and transfer the most promising 18F-labeled S1PR1 tracer into human investigations. 2) Implement PET imaging human studies in healthy controls and MS patients. When the completion of this application, a clinical suitable S1PR1 18F-radiotracer will be identified and ready to implement for MS and other inflammatory diseases.

NIH Research Projects · FY 2025 · 2017-09

The goal of the Pilot Projects and Trans-Network Activities Core is to support the development of research projects using PDX models that will advance the mission of the PDTC and PDXNet. The core will help to foster collaborative, translational research both within the PDTC and across the PDXNet by establishing two annual $50,000 awards. The goal of these supported projects will be to support the translational goals of the PDTC to advance translational research through the use of PDX models or, research methods to enhance experimental validation and reproducibility across PDTCs and the PDXNet. Pilot projects may support the development and integration of new technologies into the PDXNet program. Collaborative, PDXNet pilot projects may also be used for Cross-Network studies to establish best practice standards for research using PDX models. The Core will also engage non-PDXNet investigators, providing collaborative opportunities to use well-characterized PDX models that recapitulate human disease and further the reach of the NCI PDXNet program.

NIH Research Projects · FY 2025 · 2017-09

Abstract Natural killer (NK) cells kill tumors and infected cells by integrating signals from two functional types of receptors, activation and inhibitory. In the mouse, many of these receptors belong to the Ly49 family, encoded by a cluster of highly related genes, discovered by the applicant’s laboratory. The highly polymorphic Ly49 receptors are related to killer immunoglobulin-like receptors (KIRs) on human NK cells as outstanding examples of convergent evolution. The applicant’s laboratory previously showed that the Ly49H activation receptor is responsible for genetic resistance of C57BL/6 mice to murine cytomegalovirus (MCMV) infections. Ly49H recognizes a molecule encoded by MCMV, in a manner independent of major histocompatibility complex class I (MHC-I) alleles. Studies from other groups suggest that in non-C57BL/6 mice, other Ly49 activation receptors that are not alleles of Ly49H appear to recognize MCMV-infected cells in vitro but their in vivo significance has not been established. Ironically, a recent publication from the applicant’s lab also show that Ly49 inhibitory receptors can provide NK cell-mediated protection against MCMV. This process depends on MHC-I alleles, via Ly49 inhibitory receptor-dependent licensing or education of NK cells, and their capacity to detect loss of MHC-I expression, termed missing-self, during MCMV infection. These studies provide the basis for understanding the association of human inhibitory KIRs and their MHC-I ligands that are also paradoxically associated with protection from viral infection, indicating that further studies may have clinical significance. But it has been challenging to study these receptors further in vivo due to the complexities of the Ly49 receptors, including their polymorphism and variegated expression. Here the applicant presents preliminary data in which expression of all Ly49s have been extinguished and can be replaced by a single Ly49 expressed on all NK cells. This technological advance will facilitate the further elucidation of how NK cells control MCMV infection. Thus, the Specific Aims of this proposal are to: 1) Study non-Ly49H activation receptors in NK cell responses to MCMV. 2) Characterize NK cells with inhibitory receptors mediating MCMV control. 3) Evaluate interactions between Ly49 receptors in control of MCMV. Thus, these studies will provide new insight into the mechanisms by which NK cells control viral infection.

NIH Research Projects · FY 2025 · 2017-09

Abstract Lymphocytes are capable of robust expansion upon antigen receptor stimulation and are one of the most rapidly dividing cells postnatally. Their proliferation capacity is critical for establishing broad antigen receptor repertoires during development and also for rapidly amplifying antigen-specific immunity upon pathogen invasion. In addition, T cells can also be expanded ex vivo for experiments or adoptive cell therapies and the generation of CAR-T cells, which have been translated into anti-cancer therapies. While such robust expansion of T cells generates many effector cells, they are short-lived and unsustainable for efficacious control of infection and cancers. The goal of the proposed study is to dissect the mechanisms by which robust proliferation is coupled to such differentiation at the molecular level. We will elucidate the gene regulatory network initiated by the transcription factor c-MYC, which is oncogenic but also critical for rapid proliferation of lymphocytes at many checkpoints, in developing lymphocytes and mature lymphocytes, and define key MYC downstream genes specifically associated with the differentiation processes. Insights from these studies and selective inhibition of the differentiation processes will disclose unappreciated tumor suppressor programs and facilitate expansion of antigen-specific lymphocytes with memory/stem-like properties for long-lasting protection against infection and cancers.

NIH Research Projects · FY 2026 · 2017-09

PROJECT SUMMARY: Children born very preterm (VPT, <32 weeks’ gestation) are at increased risk for social competency impairments compared to their term-born peers, including social communication deficits and poorer quality peer relationships leading to peer victimization and social rejection. Further, internalizing disorders are 2- 3 times higher in VPT children, with symptoms evident in early childhood and lasting into adulthood. Critically, how social competence impairments evolve and persist into adolescence, and the extent that known risk factors associated with VPT birth, including psychosocial adversity and executive dysfunction, shape their trajectory to increase risk for internalizing disorders remains unknown. Additionally, the cerebellum is a key structure adversely impacted by VPT birth, and aberrant cerebellar development is now recognized as a major risk factor for social communication deficits. Altered cerebellar development, along with its functional interactions with the Frontoparietal and Default Mode Networks involved in socio-emotional function, may be a key mechanism linking VPT birth, social impairment, and internalizing disorders. We posit that individual-specific patterns of cortico- cerebellar connectivity and related white matter tract (i.e., cerebellar peduncle) development underlie the evolution of social competence deficits, and in turn, internalizing disorders that persist or worsen at age 14-15, a critical period when peer interactions become more sophisticated and central. This process begins during infancy and extends to adolescence, with psychosocial risk playing a pivotal modifying role. Advances in MRI methodology now enable characterization of structural and functional brain networks with unparalleled spatial and temporal resolution. We couple innovative MRI techniques with detailed social-interaction and psychiatric assessments in a unique, longitudinal cohort of VPT and term-born adolescents (N=302; 137 VPT and 165 term- born) with high rates of psychosocial adversity now aged 14-15 years. The cohort has been followed since birth, undergoing prospective high-quality neonatal and school-age structural and functional connectivity MRI scans and longitudinal assessments of social competence, socio-emotional functioning, and psychosocial and familial risk at ages 2, 5, and 9-10 years (>80% retention at all waves). Continued evaluation of this valuable cohort provides an unprecedented opportunity to determine: 1) trajectories of social competence and internalizing symptoms for better identification of adolescents at greatest risk for psychopathology and 2) how trajectories of cortico-cerebellar structural and functional connectivity underlie these deficits. State-of-the-art MRI acquisition and analyses will characterize the deleterious effects of VPT birth on cortico-cerebellar connectivity into adolescence. We will delineate links between cortico-cerebellar connectivity, social competence, and internalizing disorders to elucidate modifiable psychosocial exposures and improve individual-level neuroimaging, behavior, and psychosocial predictive models that inform the timing of preventative interventions during sensitive periods of brain and socio-emotional development to mitigate risk for internalizing disorders.

NIH Research Projects · FY 2025 · 2017-09

Abstract Over the last decade, genomic studies have revealed the landscape of mutations that appear in Acute Myeloid Leukemias (AML). In each case, one of these mutations represents the initiating even for that tumor. Initiating mutations can create a fitness advantage when they occur in hematopoietic stem/progenitor cells (HSPC), resulting in clonal hematopoeisis, a state that increases the likelihood of leukemic transformation. Although we have characterized some consequences of these initiating mutations, including transcriptional changes and focal hypomethylation phenotypes for the DNMT3A mutations, our understanding of how these changes promote AML is largely incomplete. Since these initiating events occur in every cell of the tumor, unlike later- acquired subclonal events, they also are attractive targets for therapy. Thus, the first goal of this research program is to define the molecular mechanisms by which initiating mutations cause AML, and to use this information to develop novel, molecularly-targeted therapies. My role will be to distill large genomic and epigenomic datasets into intuitive, comprehensible models, generating testable hypotheses about the process of cellular transformation into AML. Doing so will require creative algorithmic development and statistical modeling, areas of bioinformatics in which I am proficient. This knowledge can then guide us in prioritizing targets and approaches for novel therapeutics. A second theme of our research program is to identify the reasons for progression of AMLs with "missing" mutations. Most initiating mutations are insufficient to produce overt AML on their own, but about 5% of AML cases appear to have no clear cooperating mutations. We will therefore use new technologies and analytical approaches to search the "dark matter" of the genome for AML-relevant genomic and epigenomic changes, using long-read sequencing to query previously unresolvable portions of the genome, whole genome sequencing to explore non-coding regions and structural variation, and algorithmic development to reveal difficult to ascertain events in AML. Because of my interdisciplinary background, I have expertise in designing algorithms and statistical models, as well as a deep understanding of the biology of AML. My training, experience, and record of productive and impactful research make me uniquely suited to push the informatics and analysis aspects of this research program forward.

NIH Research Projects · FY 2024 · 2017-09

Project Summary Environmental exposures play a leading role in the development of human disease and are therefore of significant interest, but measuring the myriad of environmental exposures encountered over a lifetime is an immense challenge. Even more complicated is identifying the molecular pathways disrupted by each environmental exposure encountered. Yet, progress on both of these challenges is ultimately fundamental to understanding the contribution of the environment to human health. The overarching mission of my research program is to address this critical need by developing and applying novel metabolomic technologies. Metabolomics is a relatively new analytical approach that is ideally suited to help address these formidable challenges because it comprehensively profiles small molecules of both exogenous and endogenous origin. In practice, however, metabolomics has not fulfilled its potential in the environmental health sciences. Its application has been severely limited due to the tens of thousands of signals detected by liquid chromatography/mass spectrometry (LC/MS) that cannot be identified. Without biochemically naming the metabolomic signals, biological inference is compromised and insights into exposure chemicals or effect mechanisms are prevented. The major goal of my research program is to overcome this barrier in metabolomics to (1) enable unprecedented exposure analysis in humans and (2) to discover toxicant effect mechanisms by using zebrafish. First, with new technologies that my laboratory has developed, we will name each signal detected by LC/MS untargeted metabolomics from human blood and the zebrafish embryo. This will constitute the human and zebrafish “reference metabolomes”. We will then develop a resource to automate identification and quantitation of each metabolite in the reference metabolomes. Second, we will screen the zebrafish reference metabolome to identify effect mechanisms of specific toxicants known to have adverse effects on only a single zebrafish organ. We will test our hypothesis that each organ’s metabolism is similarly disrupted by some toxicants, but that their unique phenotypic responses reflect organ-specific sensitivities to particular pathways. Thus, in addition to building a metabolomics resource that will greatly enhance exposure analysis in human subjects, we also expect that the application of our platform to zebrafish will identify toxicant effect mechanisms and establish biochemical pathways that contribute to particular phenotypes.

NIH Research Projects · FY 2025 · 2017-09

(PLEASE KEEP IN WORD, DO NOT PDF) Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. Understanding kidney disease relies upon defining the complexity of cell types and states, their associated molecular profiles, and interactions within tissue neighborhoods. We pioneered the development of novel tissue processing methods in the KPMP that uses a limited amount of human kidney biopsy tissue for single nucleus (sn) omics and spatial technologies across institutions while allowing parallel histological evaluation. Using our TISAC-approved snRNA-seq assay on reference, AKI and CKD biopsies, we generated a snRNA atlas from 200K nuclei covering cortex to papilla and revealed 53 healthy and 47 altered cell states. These represent cycling, degenerative, adaptive/maladaptive and transitioning cells in tubular and interstitial compartments. We augmented these efforts by integrating these data with newer technologies that measure RNA and epigenome in the same nucleus and defined key transcription factors associated with gene expression changes in transition of healthy cell to altered cell states. In a collaborative effort in KPMP and HuBMAP, we integrated snRNA-seq, scRNA-seq, SNARE-seq2, two different spatial transcriptomic technologies and 3D cytometry. From this we were able to define biologically meaningful cell-cell interactions, cognate molecular interactions and genes associated with adaptive states that correlate with worse CKD outcomes. Deeper sampling and integrated analyses ensuring tissue economy of AKI and CKD biopsies is still needed to overcome gaps in clinicopathological-molecular determinants, and to assess effect of race, sex, ethnicity, clinical attributes, mechanisms of gene regulation, shifts in cell states, and characterization of biologically relevant niches. To this end we will use paired snRNA-seq and snATAC-seq (chromatin accessibility) to define mechanisms associated with altered state transitions, which will enable discovery of potential causative epigenetic factors driving RNA or protein changes in disease (Aim 1). To further define the spatial contexts of injury niches for biological insights, we will evaluate and implement highly multiplexed spatial transcriptomics technologies (Aim 2) and integrate knowledge from our transcriptomic and epigenomic atlas with other orthogonal technologies in the KPMP and other consortia (Aim 3). This collective effort will enable the creation of a high resolution spatial molecular atlas of healthy and diseased kidneys that can be used as a benchmark to interrogate and interpret molecular information in a single patient’s biopsy that can inform clinical care.

NIH Research Projects · FY 2025 · 2017-09

Project Summary/Abstract Hematopoietic cell transplantation (HCT) is the only curative therapy available for many hematological malignancies as well as some non-malignant diseases such as hemoglobinopathies, autoimmune diseases, and inherited metabolic disorders. Key obstacles to the success of HCT include collecting optimal numbers of hematopoietic stem cells (HSCs) for HCT and/or gene therapy, toxicities due to nontargeted chemotherapy- and irradiation-based conditioning, control of graft-versus-host disease (GVHD) after allogeneic HCT (allo-HCT) and treating disease recurrence both before and after HCT. I have focused my career over the last 30 years on overcoming these obstacles to HCT using a translational research approach. In the last project period, I designed, led, and/or completed correlative studies for 15 clinical trials, including studies that led to the FDA- approval of ruxolitinib (JAK1/2 inhibitor) for the treatment of steroid refractory GVHD in 2019 and motixafortide (CXCR4 inhibitor) plus G-CSF for HSC mobilization for autologous HCT in multiple myeloma (MM) in 2023. I completed first-in-human clinical trials of the bispecific antibodies Flotetuzumab and AMV564 in patients with relapsed or refractory (r/r) AML, treated the first patient with multiple myeloma (MM) with our home-grown CS1 chimeric antigen receptor T cell (CART) produced in our new GMP facility, developed an ‘off-the-shelf’ universal allogeneic CART targeting CD7 (UCART7) that lacks CD7 and TRAC to minimize the risk of fratricide and GVHD, respectively, and completed a global (12 sites; 7 domestic and 5 international) Phase I trial testing UCART7 in patients with r/r T-ALL/LBL that led to an upcoming Phase II registration trial. I also discovered a novel conditioning strategy combining CD45- or cKit-targeted antibody-drug conjugates (ADCs) with JAK1/2 inhibitors to enable robust donor engraftment in fully major histocompatibility complex (MHC)-mismatched mice. My research program over the next seven years will continue to use our strengths in preclinical modeling, cancer genomics, CART immunotherapy and the design and execution of early phase clinical trials to 1) develop novel very late antigen 4 (VLA-4) inhibitors for HSC mobilization, gene therapy and GVHD prevention, 2) test if targeting human CD45 or cKit with ADCs or human CD47 and cKit with naked antibodies, inhibitors, or bispecific molecules in the presence of JAK1/2 inhibitors can serve as effective conditioning regimens in humanized mouse models and primates, and 3) develop novel CART against T-ALL, AML, and MM. Three obstacles to successful CART therapy are: 1) limited CART expansion and persistence, 2) CRS/ICANS, and 3) on-target/off-tumor toxicities. I will test if long-acting cytokines of IL-7, IL-15 and/or IL-21 enhance CART expansion, persistence, and function. To address toxicity, we will continue testing duvelisib in humans and define the mechanism/s whereby PI3K-γ,δ inhibitors, JAK1/2 inhibitors and CD40L/CD40 blockade reduces CRS/ICANS without inhibiting CART. To minimize on-target/off-tumor toxicities, we will engineer epitopes on CART and/or donor HSCs used in allo-HCT to provide selective resistance to CART without altering the function of the edited protein.

NIH Research Projects · FY 2026 · 2017-08

PROJECT SUMMARY ABSTRACT Stroke is a leading cause of death and disability, with dismal 1-year outcomes for survivors. Many are rehospitalized, fall, are admitted to skilled nursing facilities (SNFs), or die. Stroke is fundamentally a chronic condition that is currently managed as an acute event. Acute inpatient rehabilitation (IR), focused on resolving impairments, does not typically address the environmental barriers stroke survivors face when they return home (e.g., stairs without railings). As a result, they leave IR without the skills to successfully reintegrate into the community. There is a critical gap in post-acute care as chronic needs of survivors are not being met during their transition from a medical to a community model of care. To address this gap, we developed a novel rehabilitation transition program, Community Participation Transition after Stroke (COMPASS), a behavioral intervention delivered in the home during the transition from IR that resolves barriers to independence using home modifications (e.g., grab bars by the toilet) and self-management (learning to resolve barriers independently). Feasibility and phase IIb trial results showed that COMPASS can reduce environmental barriers in the home and may impact health outcomes such as SNF admission and death. We now propose a hybrid type 1 randomized effectiveness-implementation trial to establish COMPASS as the missing link between IR and home. Our central hypothesis is that COMPASS will be superior to usual care for key outcomes and implementable in practice. The specific aims are: (1) conduct a multicenter randomized controlled trial to evaluate the impact of COMPASS on SNF admission, death, rehospitalization, and falls compared to enhanced usual care; (2) evaluate the quality, cost, and efficiency of COMPASS during transition from IR to home relative to enhanced usual care; and (3) conduct a mixed-methods study informed by the Practical Robust Implementation and Sustainability Model (PRISM) and the Reach, Effectiveness, Adoption, Implementation, and Maintenance (RE-AIM) framework to evaluate the sustainability and implementation potential of COMPASS. Hypotheses are: (1) COMPASS participants will have significantly lower incidence of 12-month SNF admissions and mortality, lower incidence of 30-day rehospitalization, and lower fall rates compared to enhanced usual care; (2) the incremental cost-effectiveness ratio will be cost-effective by World Health Organization standards, and return on investment will be positive; and (3) COMPASS will have high reach, adoption, implementation (fidelity), and maintenance (12-month adherence) and a high potential for implementation and sustainability. COMPASS is innovative because it augments current practice by resolving barriers in the environment as stroke survivors transition home. This proposal is significant as it fills critical gaps in rehabilitation evidence by investigating the therapeutic efficacy, cost-effectiveness, and implementation potential of a novel behavioral intervention during the transition from IR. COMPASS could effectively bridge the gap between acute and community care, reducing stroke survivors' morbidity and mortality.

NIH Research Projects · FY 2026 · 2017-08

7. PROJECT SUMMARY The goal of the proposed MIRA-funded research portfolio is to discover how dynamic interactions between individual biomolecules at the nanoscale influence their collective function and organization in complex biophysical processes. The proposed research program integrates continued development of 6D single- molecule (SM) imaging (3D positions and 3D orientations) with mechanistic studies of the organization of biomolecular interactions at the nanoscale. Importantly, the proposed scientific goals synergistically spur the development of impactful imaging capabilities, and these new capabilities will in-turn overcome barriers to enable novel significant scientific trajectories to be pursued. Four broad research thrusts will be pursued. Thrust 1 will develop smart adaptive 6D nanoscopy. Previous studies have shown that fixed imaging systems cannot measure all possible molecular rotational motions with the best-possible quantum-limited precision. Thus, dynamic illumination and fluorescence modulation hardware will be integrated to enable the imaging system to adapt as data is collected. Fusing model-driven design algorithms with data-driven deep learning methods will yield smart microscopes that enable measurements that are not possible even with current state-of-the-art nanoscopes. Thrust 2 will develop high-speed 6D SM tracking to map spatial heterogeneities in molecular interactions between biomolecules. These heterogeneities govern important processes like phase separation, but current techniques have sufficient spatiotemporal resolution to resolve mechanistic details. Time-varying illumination, single-photon counting, and direct pupil imaging will be integrated to visualize these dynamics using 10x fewer emission photons and thus 10x faster speed than state-of-the art methods. Thrust 3 will leverage developments in 6D nanoscopy to elucidate dynamic molecular architectures of self- assembling peptides and natural amyloidogenic proteins. Critically, scientists must disentangle the effects of peptide sequence, secondary structure, assembly architecture, and aggregation conditions to create new biomaterials for diagnostics and therapeutics, as well as to elucidate the mechanisms of cytotoxicity in amyloid diseases. The 6D positions and orientations of transiently binding fluorophores will visualize the dynamic organization of individual peptide assemblies both in vitro and as they interact with living cells with nanoscale resolution. Thrust 4 will leverage developments in 6D SM tracking to visualize heterogeneous network architectures within biomolecular condensates that ensemble measurements fail to detect. The 6D positions and orientations of fluorogenic probes will be used to characterize the network architecture of stickers and spacers within the condensate, thereby visualizing the driving forces of phase separation. Six-dimensional SM nanoscopy will also directly observe how proteins are recruited and reorganized throughout the phase separation process, leading to mechanistic insights into the formation and spatiotemporal evolution of biomolecular condensates.

NIH Research Projects · FY 2025 · 2017-08

Uromodulin-associated genetic chronic kidney disease, or autosomal dominant tubulointerstitial kidney disease caused by uromodulin mutations (ADTKD-UMOD), is a leading hereditary kidney disease and characterized by renal fibrosis and progressive loss of kidney function. Currently there is no targeted therapy. To address the unmet medical needs, by using CRISPR/Cas9, we have developed an ADTKD-UMOD mouse model carrying Umod p.Tyr178-Arg186 del, the mouse equivalent of the most prevalent human mutation. Uromodulin (UMOD) is mostly synthesized and secreted by the tubular cells of thick ascending limb (TAL). Linked with mutant UMOD-triggered endoplasmic reticulum (ER) stress, our mouse model shows that autophagy and mitophagy deficiency in TALs leads to increased accumulation of the toxic mutant UMOD aggregates and mitochondrial DNA leakage-induced activation of innate immune signaling molecule STING (stimulator of interferon genes), which cause TAL cell death and renal fibrosis in ADTKD. Most importantly, we have discovered a novel ER soluble protein mesencephalic astrocyte-derived neurotrophic factor (MANF), which functions as a positive regulator of autophagy/mitophagy. MANF tubular overexpression can decrease mutant UMOD accumulation and counteract STING-induced inflammation, thus attenuating fibrosis and improving kidney function in our mouse model of ADTKD. Our hypothesis is that restoring suppressed autophagy and mitophagy by targeting MANF and its receptor is a novel therapeutic strategy to treat ADTKD-UMOD. The overall goals of this proposal are to determine the molecular mechanisms underpinning the biological and therapeutic functions of MANF, to develop MANF-based therapy, and to define the role of newly identified MANF receptor neuroplastin in ADTKD. To accomplish our research goals, we have assembled an interdisciplinary team including multiple Co-Investigators and cores with various innovative technologies and required expertise. The proposed study will provide critical insights into the molecular pathogenesis of ADTKD and develop highly-targeted and mechanism-based novel treatments for ADTKD patients.

NIH Research Projects · FY 2025 · 2017-08

PROJECT SUMMARY TOP-TIER (Training OPportunities in Translational Imaging Education and Research) is a clinician scientist post-doctoral training program at Washington University (WU) in St. Louis designed with the purpose of providing trainees with instruction in the performance of rigorous translational imaging research. The goal is to prepare resident and fellow trainees for careers as successful independent investigators and to ultimately become leaders in their field, developing imaging techniques and applications that translate into human subjects and impact healthcare. Precision Medicine is an approach to disease treatment and prevention that takes into account individual variability in genes, environment, and lifestyle. Development of methods of patient-specific biomarker imaging to guide and monitor patient-specific therapies will be needed to advance Precision Medicine. Moreover, a trained workforce is key to taking advantage of new developments in machine learning and artificial intelligence which can be applied across disciplines, to include targeted molecular imaging, photoacoustics, and magnetic resonance imaging (MRI) in terms of data acquisition, image processing, and diagnosis. Such imaging advances are interdisciplinary with technology innovations crossing the physical, biological, and data sciences. These studies start with preclinical mechanistic inquiries using targeted pathology-based imaging in animal disease models, and then expand into human subjects. Human studies include safety testing and the Food and Drug Administration’s (FDA) Investigational New Drug (IND) and Device Exemption (IDE) process. The ultimate goal is to make research innovations widely available to the public and the entire practicing medical community. Although this “bench-to-bedside” process has been widely acclaimed, there remains a knowledge gap in the medical imaging research community as to how to take preclinical research into humans, and how to then take these innovations to the public. Moreover, clinical scientists – residents and fellows – often lack the understanding to navigate research regulatory requirements as well as the knowledge base to understand or perform preclinical research that will inform the mechanism of innovation. In this T32 renewal, we have optimized training to provide supportive Mentoring Teams and cover technical advances, research rigor, and the practical aspects of grant submission to better prepare future imaging scientists for the challenges of performing science which impacts healthcare.

NIH Research Projects · FY 2025 · 2017-08

Project summary Brainstem vestibulospinal neurons receive information about head movement from vestibular afferents, but for unexplained reasons, during locomotion they also receive a copy of motor commands, in the form of a rhythmic oscillation aligned with the locomotor cycle. It is unknown whether the job of this locomotor copy, known as a corollary discharge, is to cancel the expected head movement information—thereby allowing vestibulospinal neurons to focus on encoding unexpected head movements—or to enhance expected head movement information, reducing the error inherent in sensory signals. Furthermore, the circuits that give rise to this corollary discharge are undefined. Because other brainstem neurons, like reticulospinal neurons, also exhibit these locomotor-driven oscillations, this corollary discharge signal is likely to be important in coordinating activity between the brainstem and spinal cord more generally. The goal of this proposal is to define the synaptic components underlying this corollary discharge, measure how sensory signals combine with corollary discharge, and identify the presynaptic sources of this signal. In addition, completion of this proposal will produce a synaptic-resolution map of the brainstem, broadly accessible to other researchers. Work will be carried out in the young zebrafish, whose transparency permits accessibility to in vivo whole-cell recordings not feasible in other vertebrates. Specific goals of this project are (1) to characterize the excitatory and inhibitory synaptic inputs underlying corollary discharge and define how they summate with sensory input; (2) to acquire and leverage connectomic data to identify candidate presynaptic partners carrying this corollary discharge signal; and (3) to determine the genetic identity and function of presynaptic inputs driving corollary discharge. This proposal relies on a combination of approaches, including whole-cell in vivo physiological recordings, anatomical and optogenetic mapping of circuitry, and serial-section electron microscopy and reconstructions. The project is conceptually innovative in that it seeks to define a larger-scale principle for ascending corollary discharge carrying specific information about the locomotor cycle. The overall contribution of this work will be to define a circuit that is widespread across vertebrates and operates at the level of coordination between the brainstem and spinal cord for effective motor control.

NIH Research Projects · FY 2025 · 2017-08

PROJECT SUMMARY Although the genome is a blueprint for making an organism, the epigenome (modification on the genome without DNA sequence change) governs the functional outcome of genes and ultimately shapes the organism. Epigenetic modification is a conserved and important gene regulatory mechanism and plays quintessential roles in genome integrity, development, environmental responses, and diseases. Despite the large number of correlative studies describing the altered modification patterns in abnormal developmental and pathological tissues, whether they are a cause or a consequence is poorly understood. This proposal dissects the molecular mechanism of epigenetic regulation and functional consequences of epigenome perturbation under developmental and physiological conditions. Specifically, we investigate i) how epigenome reprogramming in response to environmental cues might lead to stress adaptation; ii) how an epigenetic switch regulates developmental phase transition; and iii) how epigenetic modification safeguards the genome integrity. We will further dissect the causal roles of epigenetic marks on gene expression and develop an innovative CRISPR/Cas9-mediated cis- engineering system for epigenome editing. Our approach is highly interdisciplinary making use of the powerful Arabidopsis genetic system, high-throughput proteomics and genomics, live-cell imaging techniques, biochemical and structural tools to tackle mechanistic problems from genome-wide scale to atomic resolution. As we probe basic principles governing epigenetic regulation that are conserved across eukaryotes, the mechanistic knowledge and epigenome technologies acquired from our pioneering Arabidopsis studies will help accelerate progress in deciphering the relevant mechanisms in human. Such knowledge will lead to the development of innovative tools to correct aberrant epigenetic modifications and ultimately enable new applications for medicine and human health.

NIH Research Projects · FY 2025 · 2017-08

PROJECT ABSTRACT. This competing renewal to Suubi4Her study (R01MH113486-Ssewamala, PI) will examine the longitudinal impact of an evidence-based combination intervention combining economic empowerment (EE) and family strengthening (FS) on HIV risk behaviors, cognitive, and mental health outcomes among adolescent girls transitioning into young adulthood in poverty-impacted and HIV-burdened rural communities in Uganda. Specifically, the study provides a unique opportunity to examine the longer-term effects of an evidence-based combination intervention on HIV prevention (for adolescent girls who are HIV negative— including PrEP use), and care and support continuum trajectories (of adolescent girls living with HIV – including ART Adherence) during transition into young adulthood, a high-risk, yet understudied developmental stage in sub-Saharan Africa (SSA). Adolescent girls and young women (AGYW) in SSA are still disproportionately affected by high rates of HIV infection than their male counterparts. In 2022, 63% of all new HIV infections in SSA were among women and girls. This increased risk of HIV infection among AGYW is exacerbated by poverty, gender inequalities, stigma, discrimination, and disruptions to cognitive development trajectories that are widespread across SSA. Yet, less than half (42%) of countries in SSA provide HIV prevention programs designed specifically for AGYW. Alarmingly, communities with high HIV prevalence also report poor mental health, including depression and suicidal ideation. This is particularly important during the transition from adolescence to young adulthood as the brain is undergoing dramatic remodeling of dopaminergic neural networks. Adolescent brain development enhances reward-seeking and peer influence, leading to risky decisions and mental health issues without proper self-regulation, which improves in adulthood. Research indicates that poor mental health during adolescence is more pronounced among girls than boys, underscoring the pressing need for girl-targeted interventions during adolescence. The World Health Organization has recommended using multi-sectoral approaches that address both structural and family-level issues. Approaches that concurrently address poverty (e.g., via EE) and poor mental health (e.g., via FS) have a potential to concurrently reduce AGYW’s vulnerabilities to HIV, and poor mental health and cognitive functioning. Our team recently concluded a 6-year (2017-2023, with one-year no cost Extension), 3-arm cluster-randomized controlled trial, called Suubi4Her, that tested a combination intervention guided by Asset Theory and family strengthening principles among 1260 school-going adolescent girls aged 14-17 years at enrollment. In this competing renewal, we propose 3 aims: Aim 1: To examine the long-term impact of the Suubi4Her intervention on young women’s HIV risk behaviors; Aim 2: To elucidate the long-term effect of the Suubi4Her intervention on cognitive and mental health, and to explore specific cognitive underlying factors that mediate the link between the intervention and reduced HIV risk; and Aim 3: To examine the long-term cost-effectiveness of the Suubi4Her intervention.