Johns Hopkins University

NIH Research Projects · FY 2026 · 2024-03

PROJECT SUMMARY Glaucoma is a major cause of blindness, affecting over 80 million people worldwide. Glaucoma is a neurodegenerative optic neuropathy caused by the loss of retinal ganglion cells (RGC), leading to loss of vision. Current therapies are all directed at lowering intraocular pressure (IOP), and yet RGC loss still continues in many patients despite IOP lowering. The identification of an agent that complements IOP lowering by promoting RGC survival would be a significant advance toward improving the visual outcomes of patients with glaucoma. Using cultures of primary RGC, we screened more than 10,000 compounds and identified candidates with potent neuroprotective properties, including a drug that is FDA-approved for an unrelated indication. We further characterized the novel pathway through which these compounds act to protect RGC, thus identifying a novel drug/drug target combination for neuroprotection. We have also developed a novel thermosensitive gel-forming eye drop drug delivery system that provides efficacious drug delivery to the posterior segment, even in large animals (rabbits, pigs). Importantly, we observed that the combination of more effective intraocular drug penetration provided by the gel-forming eye drop with a drug that binds to melanin in the eye, led to RGC protection in vivo with once weekly topical dosing. In this application, we are screening additional neuroprotective drugs for melanin binding, cell uptake, and intraocular penetration to compare head- to-head for pharmacokinetics and efficacy in an optic nerve crush rat model and a bead injection mouse model of IOP elevation. The goal is to develop an efficacious eye drop for neuroprotection that requires once weekly, or ideally once monthly maintenance dosing. In Aim 1, we will make further formulation changes in the eye drop to increase intraocular drug absorption, and formulate additional melanin-binding neuroprotective drugs. In Aim 2, we will characterize the pharmacokinetics and efficacy of a variety of dosing regimens and formulations to identify the most effective dosing approach with the lowest dosing frequency. In Aim 3, we will perform dose-ranging studies in rabbits to achieve similar drug concentrations in target tissues that were shown to be effective in Aim 2. We will also perform safety evaluations with longitudinal dosing, including fundus exams, IOP, and retinal morphology analyses. The demonstration of efficacy in rodent models of neurodegeneration along with similar pharmacokinetics and no overt toxicity in the rabbit eye, would provide evidence of the therapeutic potential of our neuroprotective drug delivery strategy for the treatment of glaucoma.

NIH Research Projects · FY 2026 · 2024-03

Project Abstract Fibrolamellar hepatocellular carcinoma (FLC) is a rare and often lethal form of liver cancer that primarily affects children and young adults without cirrhosis. There are no approved systemic therapies for FLC, and it is usually refractory to treatment approaches developed for other forms of liver cancer. A chimeric transcript between DNAJB1 and PRKACA was identified as a signature genomic event in FLC and leads to activation of PKAc. However, pharmacological inhibition of PKAc for FLC with traditional small molecule inhibitors has been infeasible due to on-target toxicity. Our preliminary data derived from preclinical models of FLC and human FLC tumors show that the DNAJB1-PRKACA fusion results in a metabolic rewiring of the tumor cell, leading to glutamine dependence. Induction of the DNAJB1-PRKACA fusion in preclinical cell lines is associated with sensitivity to glutamine antimetabolite therapy. Glutamine dependency in FLC results in a nutrient-depleted tumor immune microenvironment (TiME) that is enriched in immunosuppressive metabolites (e.g., ammonia, acidosis), impairing antitumor immunity. In an in vivo model of FLC, the combination of glutamine antimetabolite therapy plus an immune checkpoint inhibitor (ICI) reverses T cell dysfunction within the tumor immune microenvironment (TiME) and induces antitumor immunity resulting in robust tumor control. We are translating these preclinical findings into a clinical trial of a glutamine antagonist (sirpiglenastat) in combination with a PDL1 inhibitor (durvalumab). In Aim 1, we will conduct a clinical trial to test the safety and clinical activity of sirpiglenastat in combination with durvalumab, in children or adults with advanced FLC. In Aim 2, we will determine whether treatment with sirpiglenastat combined with durvalumab suppresses glutamine-dependent processes and increase the number of activated FLC-specific T cells within the tumor microenvironment. In Aim 3, we will identify the molecular mechanism and specific metabolic perturbations through which the DNAJB1-PRKACA fusion induces glutamine addiction and immune suppression. This work will advance a promising new treatment approach for advanced FLC, a tumor type that currently confers a median survival of only one year. Uncovering the activity of specific pathways that make FLC glutamine dependent will establish a more complete understanding of metabolic biomarkers of glutamine addiction, and will reveal synergistic vulnerabilities, which may be targetable for even more effective treatment approaches. We anticipate that these avenues of inquiry will likely be generalizable to other classes of glutamine addicted tumors.

NIH Research Projects · FY 2026 · 2024-03

Project Abstract Radiotherapy (RT) is used in the treatment of over 50% of cancer patients. Despite major technological developments, RT is still critically limited by normal tissue toxicity, and curative treatment is restricted to localized tumors. Significant gaps also exist in access to RT in the USA and in global health due to treatment costs, distance-time barriers, cultural and infrastructure/technology barriers. These challenges are also fundamentally reflected in the radiation oncology professional training capacity. There are now emerging transformative RT research areas and initiatives that can address the toxicity limitations in substantial ways, extend RT to curative treatment of both local and metastatic disease, and improve access and efficiency with innovations to reduce treatment time and cost, enhance patient convenience, and significantly increase survival and quality of life for patients. Some of the exciting emerging transformative areas of RT research include: Radioimmunotherapy, FLASH RT, Artificial Intelligence (AI) in RT, smart RT biomaterials, MRI-guided RT, and new technology and approaches relevant to global oncology, all with tremendous potential to engage students/trainees and faculty to advance innovation, while improving access and clinical impact. These emerging areas are exciting and there is great need for concerted and accelerated efforts to train the next generation of research leaders in these areas. To address this unmet need, the overall goal and innovation of this project is to establish a premier CaREER (Cancer Research Education Excellence in Radiotherapy) program to train the next generation of scientists, technologists and physicians skilled in research in emerging transformative areas of RT, with an innovation focus on improving access, efficiency, and clinical impact in radiotherapy. The proposed CaREER program will leverage a strong institutional environment at Johns Hopkins, in collaboration with faculty at Howard University and other U.S.-based academic partners, as well as select U.S.-based global health research programs, to broaden participation in advanced cancer research training and prepare trainees to pursue careers and advanced education in cancer research. The highest significance of the CaREER program is expected in the concerted training of next generation leaders in the newly emerging and transformative research areas of radiotherapy, to drive innovation in overcoming current RT limitations and expanding access to high-quality radiotherapy care.

NIH Research Projects · FY 2026 · 2024-02

Venous thromboembolism (VTE), consisting of deep vein thrombosis (DVT) and pulmonary embolism (PE), affects approximately 1 in 200 hospitalized patients < 21 years old, and have also generated published evidence on several subpopulation-specific RAMs over the past 3 years. Yet, critical knowledge gaps remain regarding RAM development in critically ill patients and the investigation of prognostic markers for clinically-important VTE outcomes, including recurrent VTE and PTS. In this K24 proposal, I will mentor six junior faculty clinical/translational researchers focused in VTE prevention and treatment, from across four pediatric disciplines (hematology, critical care medicine, hospital medicine, pharmacy) and three academic institutions (Johns Hopkins University [JHU], University of Alabama at Birmingham, Harvard University). The proposed studies build upon existing collaborations in data science and biomarker-informed prognostic modeling and will leverage data and/or biospecimens from multicenter studies in which I play a leadership or senior collaborating role. In each project, the junior faculty mentee will also have direct interaction with new or existing patients on study, for de novo data collection. Under this K24 proposal, I will enhance my mentorship expertise via the JHU Master Mentor Program and grow my knowledge in both Bayesian methods and plasma proteomics through online coursework and regular interactions with experts in these fields, who also serve as co-investigators in the proposed projects. By facilitating the expansion of my interdisciplinary research in pediatric VTE and strengthening my abilities to successfully mentor future junior faculty clinical and translational researchers in the field of pediatric VTE from across a range of disciplines and institutions, the proposed K24 will have significant impact as a force-multiplier in the pediatric VTE field

NIH Research Projects · FY 2026 · 2024-02

Project Summary Multiple sclerosis (MS) is a common inflammatory and neurodegenerative disorder. In MS progression of disability is irreversible, and prognosis is highly variable; some individuals rapidly progress to a disabled state whereas others experience only mild symptoms. However, mechanisms contributing to the observed heterogeneity in disease evolution are poorly understood. Discovery of novel biomarkers associated with risk of disease progression will not only allow for more accurate individualized prognosis but also facilitate the discovery of new therapeutic targets that may be relevant for targeting the progressive aspects of MS. Metabolomics is an ideal technology for biomarker discovery; an individual’s metabolic phenotype incorporates multiple levels of biologic interaction (e.g., endogenous metabolism, the exposome, and activity of the gut microbiota). We previously found robust metabolic alterations in people with MS when compared to healthy people in a study including nearly 1000 metabolomic profiles. Results suggest a marked disruption of multiple amino acid pathways, with notable reductions in metabolites related to aromatic amino acid metabolism (phenylalanine, tryptophan, and tyrosine). Lower levels of these and other metabolites were also strongly correlated with disability levels at a single time point. The overall goal of the proposed studies is to build upon these initial results by evaluating whether certain metabolic changes predict MS prognosis and explore potential contributing mechanisms using a data-driven approach. We will evaluate, in a prospective design, whether (1) certain metabolic changes predict subsequent MS prognosis in Aim 1; (2) characterize potential contributing mechanisms by considering the mediation by MS disease modifying therapies in Aim 2; and (3) assess the added predictive value of metabolomic markers when combined with traditional measures of disease severity in Aim 3. Our central hypothesis is that metabolic changes, both in in AAA metabolism, as well as other novel pathways, strongly predict subsequent clinical and radiological MS outcomes (i.e., MS prognosis). To evaluate this hypothesis, we will use data and samples collected from nearly 1500 PwMS participating in three randomized studies. These datasets offer an abundance of advantages in evaluating metabolic predictors of MS outcomes. For example, they are large cohorts in which standardized collection of biospecimens and rigorously assessed outcomes are collected at pre-specified longitudinal intervals. These valuable resources will be combined with validation in clinical, real-world NIH-funded observational cohorts of 400 PwMS. Lastly, our study will apply an innovative analytic strategy applying advanced epidemiological modeling tools rooted in causal inference. The collective results of this study stand to (1) provide novel insight into underlying mechanisms contributing to disability accumulation in PwMS; and (2) identify novel therapeutic targets that are relevant to progressive aspects of MS for which there are few treatments.

NIH Research Projects · FY 2025 · 2024-02

Heart failure with preserved ejection fraction (HFpEF) is the fastest growing form of heart failure, and is characterized by severe exercise intolerance (EI), exertional fatigue, disability-associated reduced quality of life, and increased mortality. The cause of the severe EI in HFpEF remains unclear, but prior reports and our preliminary data suggest that impaired skeletal muscle energy metabolism may contribute. Previously, our group demonstrated that HFpEF patients experience a rapid decrease in skeletal muscle high energy phosphates (HEP) during exercise, as detected with non-invasive phosphorus magnetic resonance spectroscopy (31P MRS). Other studies found that HFpEF patients have decreased skeletal muscle oxygen delivery and consumption compared to controls. However, due to methodological limitations, it is unknown whether this is due to primary impairments in mitochondrial oxidative metabolism, or whether HFpEF patients have attenuated peripheral blood flow that secondarily limits mitochondrial oxygen utilization. With the need for better in vivo methods to answer this important research question, we recently developed a novel interleaved MRS/MRI tool to simultaneously measure muscle metabolism and peripheral blood flow. Moreover, new clinically available metabolic modulators such as sodium-glucose cotransporter-2 inhibitors (SGLT2i) have been shown to improve clinical outcomes, but their impact on muscle metabolism in HFpEF has not been studied or related to EI. Finally, conventional EI measures during laboratory exercise testing fail to account for activities of daily living or sedentary behavior, but these can now be measured with recent advancements in wearable health technology. However, the relationship between these measures and skeletal muscle energetics have not been investigated in HFpEF patients. Therefore, we will leverage our new MRS/MRI tool to test the central hypothesis that abnormalities in skeletal muscle HEP metabolism are closely linked to manifestations of EI and fatigue in the daily lives of HFpEF patients and can be attenuated with new metabolic modulators. The specific aims are: (1) optimize and refine our novel interleaved MRS/MRI tool and investigate whether rapid HEP decline during exercise occurs despite preserved blood flow in HFpEF patients, (2) explore whether metrics of activities of daily living are closely related to conventional measures of EI and muscle metabolism in HFpEF, and (3) investigate whether SGLT2i administration improves muscle metabolism and reduces EI in HFpEF. The combination of these three elements will give the PI vital experience in developing clinical MRS/MRI research tools, evaluating wearable health device data, and conducting a clinical longitudinal study that will generate crucial preliminary data for a future randomized controlled trial using metabolic modulators. This Pathway to Independence award will be supported by excellent career development resources at Johns Hopkins and mentorship from experts in MR, metabolism, heart failure, wearable technology, and clinical trial design. The new tools and approaches will provide novel insights into EI in HFpEF as well as transferable skills that the PI can leverage in his future research endeavors.

NIH Research Projects · FY 2025 · 2024-02

Project summary Parkinson's disease (PD) is a prevalent and progressive neurological disorder impacting more than 1 million Americans with a well-known prodromal period in which individuals experience non-specific symptoms before receiving a diagnosis. Critically, this diagnosis often occurs after widespread, irreversible neurodegeneration has already occurred. Those with PD then encounter an escalating burden of disability and symptom severity, resulting in substantial direct medical expenses related to pharmaceutical treatments, hospitalizations, and skilled nursing care. Despite the significant disease burden associated with PD, there is a scarcity of effective preventive measures or risk reduction strategies, and limited understanding of factors that influence the rate at which clinical PD develops or ‘phenoconversion’ for those with prodromal symptoms. Inflammation is an emerging key disease process contributing to PD risk and may play a key pathogenic role during the prodromal period. Conditions characterized by chronic inflammation and immune dysfunction, including several autoimmune disorders, are considered potential risk factors for PD; certain immune agents used to treat these conditions have also been associated with a lower risk of PD, offering a novel approach to reduce PD risk by targeting immune processes. However, the results of existing are inconsistent, potentially due to inadequate control for confounding or bias related to reverse causation; few have also assessed the impact of immune factors on the rate of phenoconversion to clinical PD. The goals of this proposal are to evaluate whether long- term exposure to immune therapies is a strategy to reduce PD risk and phenoconversion rates. For this study, we will leverage genetic and risk factor information and detailed follow-up available as a part of three large-scale biobanks in which we will consider comprehensive array of therapeutic agents used to treat a range of autoimmune diseases (AIDs) as they relate to PD. Specifically, we will utilize the well-characterized effects of genetic variation in influencing the expression or function of individual AID therapeutic targets as instruments to help to understand the downstream effects of AID treatment on PD risk and phenoconversion. This epidemiologic framework will be embedded within a comprehensive bioinformatic pipeline in which extensive differential expression and epigenetic remodeling-based analyses at the single-cell level will further refine and characterize novel immune-associated therapeutic targets in the context of PD. Our central hypothesis is that inflammatory mechanisms contribute to both PD and AIDs, and that certain therapies approved to target immune processes in AIDs may be novel strategies to mitigate PD risk and phenoconversion. The collective results of this project will (1) help to improve risk stratification; (2) unravel new mechanisms contributing to disease risk and progression in PD’s early stages; and (3) aid in the design of future trials testing novel therapeutic targets for which there are existing agents with a strong biological foundation.

NIH Research Projects · FY 2026 · 2024-02

Project Summary Improved point-of-care tests and diagnostic algorithms for tuberculosis (TB) are urgently needed to enable more timely and accurate diagnosis. Currently, lack of diagnosis and diagnostic delays are significant contributors to increased mortality [1,2,3] in low and middle-income countries (LMICs), due to reliance on insensitive, slow and/or locally inappropriate tests and limited access to optimal diagnostic modalities. Fortunately, rapid advances in diagnostic imaging technology have produced affordable, portable, point-of-care ultrasound devices that can be transported with exceptional ease to resource-limited settings. Lung ultrasound (LUS) is now regularly used to accurately diagnose a variety of pulmonary disorders including pneumonia and pulmonary edema. Our preliminary studies have demonstrated 96% sensitivity of LUS for detecting associated sonographic abnormalities in patients with microbiologically confirmed pulmonary tuberculosis (PTB).5 However, there remain barriers to implementing LUS for TB diagnosis, including a scarcity of robust data about ideal training and scanning procedures. Our overall goal is implementation of real-time AI-facilitated LUS for timely evaluation of people with TB-suspected symptoms to triage who needs further evaluation and testing. Our preliminary data suggest LUS may be highly sensitive for the diagnosis of PTB, but no rigorous, adequately powered studies have investigated lung ultrasound findings in patients with PTB versus controls without PTB. To address these critical information gaps, we aim to: Aim 1. Develop a LUS model for PTB detection as a triage tool for PTB diagnosis which can be used in field settings in low-resource and remote areas. Hypothesis: LUS will have similar or better sensitivity for diagnosis of PTB compared to CXR with moderate (i.e., 70-80%) specificity when interpreted by trained personnel and validated by experts. Aim 2. Develop and test an artificial intelligence (AI) algorithm for detecting PTB by LUS that does not require trained personnel for use in LMICs. Hypothesis: An AI algorithm based on convolutional neural networks (CNNs) will classify LUS features indicative of PTB with high sensitivity (90)% vs. the reference standard of microbiological testing. This study will leverage resources and expertise among partners in the United States and Peru. Our multidisciplinary research team at the Universidad Peruana Cayetano Heredia (UPCH), the Peruvian NGO A.B. PRISMA, and the Johns Hopkins University have a strong track record of collaborative work in novel research projects in resource-poor settings, including TB and LUS. The use of portable AI-augmented LUS could save lives in resource-limited settings by decreasing time to case detection and treatment initiation.

NIH Research Projects · FY 2026 · 2024-02

Abstract Immunotherapies for cancer, including Adoptive Cell Transfer (ACT), chimeric antigen receptor (CAR) T cell therapy, and Immune Checkpoint Blockade (ICB), have seen great clinical success. However, they are still limited by cost, variable efficacy and resistance, and lack of quality targets. Technologies to track immune responses, and in particular tumor-specific T cell responses, would help researchers and clinicians to better understand variable responses in patients, design better targets for therapies, and track patient health outcomes. In this proposal, we aim to adapt our novel hydrogel-based immune cell expansion system to detect and characterize rare, neo-epitope specific T cells in the peripheral blood. This platform, termed the artificial T cell stimulation matrix (aTM), is a hyaluronic acid hydrogel conjugated with signals 1 (peptide-MHC), 2 (anti-CD28) and 3 (cytokine support). In order to create an aTM-based high- throughput detection system, we will first investigate the effects of physical (i.e. stiffness) and biochemical (i.e. activation molecules and cytokines) cues of the aTM on the expansion of neo- epitope specific anti-tumor T cells in mice. We will test the optimized aTM for batched expansion from bulk murine splenocytes in order to create a high-throughput system. Next, we will develop a process to detect and expand tumor-associated antigen-specific T cells from peripheral blood of healthy donors and compare this to current gold standards of antigen-specific T cell detection. Finally, because disease status may impact our ability to detect rare T cells, we will verify that the system can be used for detection of neoepitope-specific T cells from the peripheral blood of melanoma patients. Access to blood from patients will also allow us to determine how ICB treatment affects the immune response to the tumor by investigation of expanded neo-epitope specific T cell phenotype and function before and after therapy. The development of aTM as a high-throughput detection system for neo-epitope, patient-specific T cell responses will allow pre-clinical and clinical researchers to study immune responses to cancer and to improve cancer immunotherapies by bringing the power of these therapies to more patients.

NIH Research Projects · FY 2025 · 2024-02

PROJECT SUMMARY/ABSTRACT Systemic lupus erythematosus (SLE) is a chronic autoimmune disease of variable severity and course. The disease is characterized by a tendency for flares, in which symptoms get worse, followed by periods of quiescence (also termed remission) that can last for months or even years. While drivers of disease activity have been extensively studied in SLE, regulatory pathways activated during remission remain largely unknown. In preliminary studies, we identified soluble Izumo1 as a potential candidate involved in long-term remission in SLE. Izumo1 is the ligand for the Izumo1 receptor (Izumo1R), which is one of the most prominent genes induced by FoxP3, and has been shown to be essential for the maintenance of FoxP3-expressing T regulatory (Tregs) cells. Izumo1R is also induced in chronically activated conventional CD4+ T cells, where it has been proposed to be a marker of hypo-responsiveness and anergy. Our overarching hypothesis is that soluble levels of Izumo1 increase as a compensatory mechanism to activate regulatory pathways that attenuate inflammation in SLE. In this proposal, we will study a large prospective cohort of patients with SLE to provide clinical and functional evidence to support or discard this novel hypothesis. If this proposal is successful, it may identify the first biomarker and potential therapy linked to the induction of remission in SLE.

NIH Research Projects · FY 2025 · 2024-02

Parkinson Disease (PD) is a debilitating neurodegenerative disease, leading to progressive motor and cognitive dysfunction that affects 1% of the population over the age of 60. Although often considered a late onset disease, several lines of evidence suggest that disease onset occurs decades before motor symptoms arise. Prodromal PD symptoms, like intestinal dysmotility, anosmia, sleep disturbances, and depression may present decades prior to the onset of motor deficits, at which point a patient has lost ≥60% of their dopaminergic (DA) neurons from the substantia nigra (SN). Therefore, this proposal addresses a pivotal question in the study of late onset neurological disease. The overarching aim of this proposal is to define the timeline along which PD- related neurodegeneration begins, versus when it begins to produce quantifiable and potentially irreversible neurological effects. We will begin addressing this question by determining whether there is a critical time interval in which a PD causative mutation mediates its pathological effect. Indeed, PD mouse models that constitutively express disease-causing mutations often yield milder phenotypes than those induced later in life. The observation of this effect in multiple independent strategies points to the potential for developmental compensation rather than a technical or experimental phenomenon. Whether expression of PD-causing mutations mediate their effects in a time dependent manner, remains largely untested. By characterizing this window of pathological effect, our proposed work has the potential to inform a new wave of research involving therapeutic strategies and early disease screening. We propose to employ a tetracycline (Tet)-inducible model of the human alpha synuclein (SNCA) hA53T PD mutation, under the control of a DA neuron regulatory sequence (PITX3-IRES2-tTA/tetO-A53T) to study this question directly. To ensure the biological robustness of addressing this question with this Tet-Off model, we will first evaluate the latency between administration of the tetracycline analog, doxycycline (Dox), and repression of SNCA A53T transcript and protein levels, as well as between Dox removal/washout and SNCA A53T de- repression. Next, to determine whether the pathological effect of mutant SNCA is mediated throughout life or within a defined time window (e.g., gestation, postnatal, juvenile, or mid-adult life), we will activate expression of the mutant protein across five time windows: between mid-gestation (embryonic day 15.5) and postnatal day 21 (P21), between E15.5 and postnatal day 60 (P60), between P21 and mid-late adulthood (8-16 mos), between P60 and mid-late adulthood (8-16 mos), and mid-late adulthood (8-16 mos). These will be compared to controls expressing the mutant protein from E15.5 onwards and those in which the mutant protein remains inactivated. We will use well-established assays to evaluate the impact on behavioral, motor, and histological phenotypes. Our proposal will significantly advance the understanding of the context (timing) during which expression of PD mutations in SNCA mediate their effect in PD-relevant cell types, including DA neurons.

NIH Research Projects · FY 2025 · 2024-02

Project Summary/Abstract Increased ribosome biogenesis is a hallmark of cancer and reflects the increased and pervasive protein synthetic needs of the cancer cells. Targeting this process by curbing the rate-limiting step, RNA polymerase I (Pol I) transcription, using specific inhibitors abrogating this activity, is a promising strategy for cancer therapy. BMH-21 is a first-in-class small molecule that inhibits Pol I transcription and large-scale cancer cell line screens demonstrated its potent efficacy across broad cancer lineages. However, a heterogeneous response was observed emphasizing the presence of mechanisms dampening the therapeutic response. Genome-wide positive selection CRISPR Cas9 knock-out screens were performed in human colorectal carcinoma cells to identify genes that cause resistance to the selective inhibition of Pol I. The screens identified all key positive regulators of the mTORC1 pathway accounting for the resistance. mTOR is a major driver of ribosome biogenesis and cellular translational programs and considered essential for cancer growth. The inactivation of mTOR as a resistance mechanism is counterintuitive, as this mechanism requires a reduced translational state by the cancer cells and inactivation of two key pathways enabling protein synthesis. These unexpected findings led to the hypothesis that cancer cells evade severe ribosome biogenesis stress by switching off mTOR-dependent translational pathways. This premise stipulates that bypass mechanisms have evolved to facilitate translation of essential proteins for survival and growth when cancer cells face obstacles in protein translation. This concept will be tested using pharmacological and genetic approaches that block mTOR. The goal of the project is to gain knowledge on the mechanisms that enable unabated ribosome translation in cancer cells. The following aims will be implemented to achieve this goal: Aim 1. Identify genes driving resistance to Pol I and translation inhibitor, Aim 2. Investigate the translational activity maintained under translation stress in cancer cells and Aim 3. Identify the regulators of ribosome activity that support survival by translation stress. By executing these aims we will gain knowledge how cancer cells survive severe translational stress and the mechanisms of this escape. These studies will identify critical processes and proteins required for survival and how their translation is maintained with limited ribosome numbers. These findings have implications in therapeutic strategies that target ribosome biogenesis, protein synthesis and translation and the approaches are designed to use this knowledge to develop new combination strategies to overcome these resistance mechanisms.

NIH Research Projects · FY 2025 · 2024-02

Detecting Reversal of Apoptosis in Cancer Cells in Mice Project Summary Targeting apoptotic pathways is one of the most important therapeutic strategies for cancer treatments, and primary cancers often exhibit dramatic initial responses to such treatments. However, most metastatic cancers inevitably recur, leading to treatment failure. The classical view of apoptosis has long assumed that, once initiated, this cell suicide process is irreversible. Challenging this assumption is our recent highly original discovery that apoptosis can be reversible, even at late stages, in multiple human cancer cell lines in vitro. We named this cell recovery mechanism “anastasis”, which means “rising to life” in Greek. Removal of a death stimulus is sufficient to allow anastasis to occur, thus indicating that anastasis is an intrinsic recovery phenomenon. We hypothesize that anastasis is an unexpected escape tactic used by cancer cells to survive cell-death-inducing cancer therapy, thereby causing cancer recurrence. If this hypothesis is true, anastasis would be a novel therapeutic target for suppressing cancer recurrence and progression. However, anastasis in cancer cells in vivo has yet to be demonstrated, and the consequences of anastasis remain undiscovered. To detect anastasis in live animals, we have successfully developed a new biosensor to label anastatic cells in fruit flies, by fluorescently tagging cells that have reversed apoptosis. This permanent tag is a unique tool for detecting and following the consequences of anastasis in vivo. Here, we will apply the similar strategy to detect and track anastasis in cancer cells in mice. By incorporating biosensor-expressing human cancer cells with three clinically relevant xenograft mouse models of cancer relapse, we will determine whether anastasis occurs in cancer cells in vivo, and if identified, whether it contributes to recurrence after anti-cancer therapy. Our results will lay a strong foundation for developing anastasis-targeting anti-cancer therapy, by generating essential animal models for studying the consequences of anastasis, and by revealing the therapeutic potential of harnessing anastasis to suppress cancer recurrence.

NIH Research Projects · FY 2026 · 2024-02

Project Summary Bacteremia, the presence of living bacteria in blood, can lead to life-threatening sepsis and represents a primary cause of death despite advances in modern medicine. For the diagnosis of bacteremia, the current gold standard is blood culture. Unfortunately, blood culture is insensitive and can take up to 5 days to complete. During this lengthy time window, patients may continue receiving ineffective or unnecessary broad-spectrum antimicrobials, leading to poor treatment outcomes, adverse iatrogenic effects, and increased selection for multi-drug-resistant “superbugs”. Molecular diagnostics promise to speed up the diagnosis and improve the treatment of bacteremia. To date, however, such a promise remains unfulfilled because existing molecular diagnostics cannot simultaneously accomplish: 1) rapid identification of species of the bacteria from a broad panel of potential causative bacteria, 2) prompt testing of the susceptibilities of the identified bacteria against various antibacterial agents, and 3) “needs-driven” versatility to adjust test specifications accordant with clinical context. A better molecular diagnostic platform is therefore urgently needed for the acute management of bacteremia. In response, we propose to develop an integrated molecular and single-cell detection platform capable of rapid bacterial detection, species identification (ID), bacterial load quantification, and antibacterial susceptibility testing (AST) in a streamlined test that allows customizable workflow. First, the proposed platform will perform bacteria ID and measure bacterial load from blood via ultrafast magnetofluidic PCR and “probe melt analysis” (PMA) within a magnetofluidic ID device in 30 min. This device is composed of a low-cost cartridge and an automated instrument. The cartridge and the instrument are designed to perform bacteria lysis, magnetic-based DNA extraction and purification, and PCR-PMA – a unique strategy that detects each bacterial species by a DNA probe with a specific melting temperature and fluorescence color code. The species ID and bacterial load provides guidance and specifications for downstream AST. Next, the proposed platform will conduct “single-cell molecular AST” to quantitatively detect bacterial ribosomal RNA as a surrogate viability marker at the single cell level, which has the potential to accelerate testing time to below the time scale of bacteria replication. Single-cell molecular AST will be performed within a microfluidic chip and a companion instrument that can automate the assay steps – including antibacterial exposure, bacteria lysis, and single-cell RT-PCR-PMA – in 90 – 150 min. The proposed molecular detection platform can provide timely and objective data to clinicians during acute care of bacteremia, which can improve their ability to establish diagnosis and administer antibacterial treatments, potentially improving clinical outcomes and curtailing the emergence of multi-drug resistant bacteria.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY Blood transfusion is the most commonly performed hospital procedure, with over 11 million red blood cell (RBC) units transfused each year in the United States alone. To accommodate time lags between blood donation and transfusion, blood units are preserved and refrigerated until use. Current FDA guidelines limit the shelf life of Adsol-preserved stored RBCs to 42 days. Beyond this window, efficacy of the transfusion product is thought to be compromised. This is because storage alters the RBC membrane permeability and eventually leads to RBC stiffening and loss of flexibility, irreversible sphericity, and a diminished ability to pass through the microcirculation once transfused. Despite standard FDA guidelines, these adverse changes are known to progress at varying rates and are thought to have a differential impact on transfusion outcomes in vulnerable populations. Whether RBCs should be transfused after a 35-day limit is a topic of debate; however, substantial evidence suggests that a tailored approach, accounting for the wide variation in RBC deterioration rates, should be implemented rather than a universal expiration date. Blood has been described as a viscoelastic fluid, with quantifiable elasticity, viscosity, and relaxation time that alter with storage age and with the proportion of stiffened RBCs. However, the deterioration rates (e.g., stiffening) of RBCs vary widely by the blood donor, and storage age alone is not a reliable indicator of RBC quality. Our group has been developing acoustic radiation force impulse (ARFI) ultrasound, a promising approach for monitoring blood characteristics through sono-transparent surroundings, such as polyvinylchloride blood bags. The advantage of a contact-free assay is two-fold: 1). Human blood is a biohazard and exposure risks should be minimized, and 2). Sampling blood units by standard techniques breaks the sterility of the blood bag, rendering the blood product unusable. A point-of-care, contact- free assay to evaluate stored RBC quality prior to transfusion has the potential to transform clinical practice by implementing a precision medicine approach to transfusion efficacy. Although shear wave elastography imaging (SWEI) is a commercially available ultrasound technique for assessing viscoelasticity of solid tissues, it is not amenable to anticoagulated blood because shear waves rapidly attenuate in fluid media. Unlike SWEI; however, ARFI does not depend upon shear wave propagation, making it more suitable for the evaluation of fluids. In this 3-year project, we will accomplish the following: 1). Optimize the ARFI beam sequences to assess stored blood viscoelascity, using ektacytometry (i.e., elongation index, a measure of bulk RBC deformability) and viscometry as comparative references for ARFI metrics, 2). Validate the clinical relevance of ARFI viscoelasticity by analyzing pre-transfusion ARFI metrics and post-transfusion hemoglobin increment, in an observational study enrolling 50 study participants undergoing clinically indicated RBC transfusion, and 3). Compare ARFI metrics versus storage age as predictors of transfusion outcomes.

NIH Research Projects · FY 2025 · 2024-02

PROJECT SUMMARY Programmed cell death (self-induced) is intrinsic to cellular life, including unicellular species. However, cell death research has focused primarily on animal models to understand cancer, degenerative disorders, and developmental processes. In contrast, there is comparably little knowledge of how prokaryotic and eukaryotic microbes die, and cell death mechanisms in human fungal pathogens (multi- or unicellular) are nearly unstudied. Over a million fungal infections are diagnosed annually, many causing significant mortality, and resistance to conventional therapies continues to increase. Building on the principles of cell death discovered in the metazoan cell death field, we propose to characterize novel cell death pathways in unicellular fungal species using the tractable Saccharomyces cerevisiae model system. Targeted genetic approaches, cell biological and biochemical approaches will be used to dissect a vesicle trafficking cell death pathway in yeast that results in lysosome/vacuole membrane permeabilization and cell death, with implications for the human pathogen Cryptococcus neoformans. This new mechanistic understanding is intended to provide fundamental knowledge needed for the future development of new therapeutic strategies analogous to successes in targeted cancer therapy by inducing intrinsic cell death mechanisms.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY: Diabetes accounts for 10% of healthcare spending in the United States. A growing portion of this cost is spent on treatments for diabetic eye disease, the leading cause of blindness among working-age adults in the developed world. Early nonproliferative diabetic retinopathy (NPDR) is driven by hyperglycemia that promotes injury to the retina microvasculature. On the other hand, proliferative diabetic retinopathy (PDR) develops after progressive microvascular damage ultimately results in retinal ischemia and subsequent hypoxia, driving the expression of factors that promote angiogenesis. Several labs have demonstrated that the regulation of angiogenic genes in late stages of diabetic eye disease is mediated by the transcription factor, hypoxia-inducible factor (HIF) in patients with retinal ischemia and PDR. However, a role for HIF in early stages of diabetic retinopathy (DR) is unclear. We recently demonstrated that transient episodes of hypoglycemia promotes the nuclear accumulation of HIF-1α, independent of hypoxia. This, in turn, results in an increase in expression of the angiogenic mediators that promote the growth of abnormal, leaky vessels in patients with DR. More recently, we have observed that activation of the p38 signaling pathway is necessary for HIF-1α accumulation in response to transient hypoglycemia. These observations implicate the p38/HIF-1 pathway in early. Events in the promotion of DR progression, independent of retinal ischemia. Based on these observations, we propose that inhibiting the p38/HIF-1α pathway will be an effective approach to prevent the progression of DR. In this proposal, we use a combination of genetic and pharmacologic approaches to evaluate the safety and efficacy of therapies that target HIF-1 (SA1) or p38 (SA2) to prevent the progression of DR.

NIH Research Projects · FY 2026 · 2024-02

Bacterial Vaginosis (BV), a disorder of the vaginal microbiota characterized by low levels of protective Lactobacilli and higher abundance of a variety of anaerobic bacteria colonizing the vagina, is the leading cause of vaginal complaints in women and associates with increased risk of preterm birth and HIV and STI acquisition. Recommended first-line BV therapies include oral or vaginal metronidazole or vaginal clindamycin, although most are treated with metronidazole; clindamycin is much less frequently used. It is well known that 60% of BV recurs within 1 year after antibiotics. What is less appreciated, however, is that a significant proportion of patients may experience poor outcomes (either persistent, refractory BV [refBV] or early symptomatic recurrence [erBV]) within 6 weeks after standard BV therapy. Defining vaginal micro-environmental factors at the time of symptomatic BV (sBV) diagnosis that associate with subsequent ref or erBV, could provide critical insights into disease pathophysiology and inform the development of candidate clinical biomarkers to predict these conditions. In the long term, this could change clinical practice by identifying a priori those for whom standard BV treatments are likely to be inadequate, who may need more aggressive initial therapy with regimens currently reserved only for those with demonstrated recurrent BV (≥3 recurrences in 12 months), [e.g. alternative or higher dose antibiotics, intra-vaginal boric acid, suppressive antibiotics, or novel bio-therapeutics currently under development.] Early data suggest vaginal microbiota features, or a disordered immune response may predict ref or erBV. Additionally, virulence factors of specific BV-associated bacteria may lead to vaginal epithelial cell (VEC) damage, revealing deeper (immature) VEC layers and resulting in decreased glycogen to support Lactobacillus growth, increasing risk of ref or erBV. Long term, deciding which adjunctive treatments to use or to develop for those at risk of poor outcomes early after therapy will depend on understanding underlying biologic mechanisms driving disease. The focus of this grant is to identify vaginal microenvironmental factors associated with ref or erBV. Our work will also establish a sample repository to be used for future studies to characterize the pathophysiology of ref and erBV. We propose to recruit a cohort of patients diagnosed with acute BV and follow them over 6 weeks to accomplish these Aims: 1. Establish a prospective cohort and specimen repository from individuals with refBV, erBV and early remission. 2. Using advanced metagenomic techniques, compare the vaginal microbiome at sBV diagnosis prior to treatment in those who experience ref and erBV vs. early remission. 3. Assess mucosal immunologic profiles, vaginal glycogen and VEC maturity at sBV diagnosis in those who experience ref and erBV vs. early remission. The study will be fully powered to assess aims in those initiating metronidazole (the most commonly used therapy) for acute BV. This research will identify candidate predictive biomarkers for ref/ erBV, and, long term, will provide data to inform mechanistic and interventional studies to improve BV care.

NIH Research Projects · FY 2026 · 2024-02

Project Summary / Abstract The goal of this grant is to develop enabling technology to address fundamental limitations in robotic intraocular microsurgery with a specific focus on high-dexterity micromanipulation. Vitreoretinal surgery may be the most technically demanding type of eye surgery and deals with the surgical treatment of retinal and posterior segment diseases. Following the trend in microsurgery, robotic assistance, enhanced by advanced imaging, has the potential to fundamentally change the field of intraocular surgery. Still in its early stages, robotic retinal surgery has been cautiously introduced into the operating room and has been successfully evaluated in a limited number of clinical trials. Owing to its demonstrated capabilities, robotic intraocular microsurgery has the potential to assist the surgeon and provide super-human physical capabilities, enabling unprecedented as well as safer surgical care for patients. Among the most relevant procedural tasks that may benefit from the use of robotic assistance is epiretinal membrane (ERM) peeling, the most common vitreoretinal surgery performed in the US. The incidence of intra and postoperative complications ranges from 2% to 30%, depending on the circumstances. The main complication is varying degrees of mechanical retinal trauma that result from accessing the membrane edge or due to excess forces applied during membrane removal. Current limitations and challenges of robotic approaches include logistically cumbersome setup, limited access to important portions of the retina, and lack of force feedback to the surgeon at the tool tip and shaft. Similar problems exist in laparoscopic surgery. To date, non-ocular robotic systems have demonstrated significant dexterity enhancement by integrating additional degrees of freedom and force-sensing capabilities at the distal end of the surgical instruments. To prove the hypothesis that a high-dexterity robotic assistant will overcome important limitations in conventional ophthalmic microsurgical procedures, we propose the following specific aims: (1) Clinically compatible high-dexterity robotic system: develop a miniature robotic forceps with snake-like distal end, and embed optical fibers-based sensors on the tool shaft, allowing force-sensing at both the tool tip and sclerotomy, and integrate the dexterous manipulator with the Steady Hand Eye Robot (SHER); (2) Control methods for high- dexterity robotic system: develop teleoperated control for the dexterous manipulator and SHER, combine teleoperated and cooperative control for the integrated system, and develop control schemes able to assist the surgeon with sensorimotor guidance for safe robotic ERM peeling; (3) Validate the high-dexterity robotic system for ERM peeling: validate the capabilities of the proposed system for ERM peeling with established ex-vivo phantoms (membrane of fertilized chicken eggs) and in-vivo biological membranes (pig eyes). This highly innovative system will fuse tool-tissue force information with intraoperative guidance via a high-dexterity robotic assistant surgical platform with cooperative and teleoperated control.

NIH Research Projects · FY 2026 · 2024-02

Abstract Huntington's disease (HD) is caused by a CAG repeat expansion in the huntingtin (HTT) gene, leading to the pathogenic expansion of a polyglutamine tract in the huntingtin protein. The key pathological features include striatal medium spiny neuronal loss, and the presence of cellular inclusion bodies containing mutant HTT proteins. There is no disease-modifying treatment for HD. The recent failure of the clinical trial of HTT reduction approaches indicates the urgent need for development of novel therapeutic modalities. Neuroinflammation has implications in HD pathogenesis. HD human tissue and mouse models demonstrate activated microglia and astrocytes and elevated levels of plasma cytokines and chemokines, all potentially contributing to HD pathology. Inflammasomes are key components of the innate immune response, whose formation is triggered by substances produced through infections, tissue damage, or other disorders. Inflammasome activation has implications in neuroinflammation and neurodegeneration. Our pilot studies found that HTT interacts with NLRP3 and activates NLRP3-inflammasome, and that knockdown of NLRP3 protects against mHTT-induced neurodegeneration in cultured human striatal neurons. These results suggest that HTT/NLRP3-linked inflammasome pathways may play a critical role in HD pathogenesis. Thus, we propose to test the hypothesis that mutant HTT interacts with and activates the NLRP3-inflammasome in neurons, thereby resulting in neurodegeneration and HD pathology, and thus that NLRP3 could be a therapeutic target via 3 aims. Aim 1. We will characterize the interaction of HTT and NLRP3. Aim 2. We will assess whether mutant HTT activates the NLRP3-inflammasome in neurons, leading to neurodegeneration. Aim 3. We will evaluate whether downregulation of the HTT/NLRP3-inflammasome pathway as a novel treatment strategy in HD mouse models. These studies will elucidate the roles of mHTT and NLRP3 interactions in activation of the inflammasome pathway and determine how this contributes to the neurodegeneration underlying HD pathogenesis. Our results may provide novel drug targets and therapeutic strategies for future HD interventions, and may also implicate a shared mechanism of inflammasome activation for other neuroinflammation-related neurological disorders.

NIH Research Projects · FY 2025 · 2024-02

PROJECT SUMMARY Pangenome references and indexes have been shown to alleviate the reference bias problem. Computer scientists recently described the novel “move structure,” which supports similar pattern-matching capabil- ities as the more typical r-index or F M -index structures, but with radically improved locality of reference. That is, move-structure algorithms access computer memory in a predictable way that minimizes cache misses, or other kinds of pauses due to data movement. We will adapt the “move structure” to the problem of pangenome indexing, enabling extremely and consistently fast pangenome queries. This will allow us to leverage inclusive and bias-avoiding pangenomes in applications where (a) we must keep up with a sequencer in real-time, e.g. nanopore sequencing, or (b) the index is so big that we must divide it across many computers, e.g. BLAST-like sequence classification.

NIH Research Projects · FY 2025 · 2024-02

PROJECT SUMMARY/ABSTRACT Anti-dsDNA antibodies are serological hallmarks of systemic lupus erythematosus (SLE), and key markers for diagnosis and disease activity. The close association of anti-dsDNA antibodies with SLE has suggested that understanding the origin of these antibodies would reveal key mechanisms in SLE pathogenesis. Nonetheless, while anti-dsDNA antibodies are likely the best-characterized autoantibodies at the genetic and molecular level, their antigenic origin and mechanisms of pathogenicity remain unclear. To date, the conclusion from previous studies is that anti-dsDNA antibodies in SLE originate from non-reactive precursors, which undergone affinity maturation against dsDNA as the primary antigen. Different to this paradigm, our preliminary studies suggest an alternative hypothesis in which anti-dsDNA antibodies are cross-reactive antibodies that originate from germline precursors targeting a protein self-antigen, and that cross-reactivity to dsDNA results from somatic hypermutation. Because gaining reactivity to dsDNA has little or no effect on reactivity to the original protein antigen, this process creates cross-reactive autoantibodies with the functional capacity to target the protein and bind dsDNA with high affinity. As a model, this new paradigm offers a rational explanation for the high heterogeneity of anti-dsDNA antibodies in terms of origin, physicochemical and pathogenic properties. To address this novel hypothesis, we will define the extended antigen specificity of a large set of mutated and germline reverted SLE-derived monoclonal anti-dsDNA antibodies using a human proteome microarray platform. The goal is to uncover both the primary specificities that preceded the origin of anti-dsDNA antibodies, and the protein targets that may determine their pathogenic effect in SLE. The final goal of this work is to gain new insights into disease mechanisms, thus laying the foundation to explore novel therapies.

NIH Research Projects · FY 2025 · 2024-01

Project Summary New experimental models are needed to develop treatments for varicella zoster virus (VZV), a DNA virus that has infected over 90% of individuals worldwide. Primary infection causes chickenpox, while reactivation of the virus within the nervous system can have dire consequences. Encephalitis is the most fearsome complication, and VZV represents one of the most common infectious causes of encephalitis worldwide. Shingles (herpes zoster) is the most common manifestation of viral reactivation, affecting one out of every three people in the U.S. and causing post-herpetic neuralgia (PHN), a severe, often chronic, pain syndrome with annual U.S. costs of over $1 billion. Importantly, VZV continues to exact a marked toll despite the advent and widespread use of antiviral agents (acyclovir) and vaccination, which only afford partial protection and do not specifically address PHN. Thus, new therapies are needed to limit the morbidity and mortality associated with neuronal infection and reactivation. The major limitation in the study of VZV infection is the strict species-specificity of the virus, as it essentially exclusively infects human cells. As a result, there are no robust animal models that recapitulate essential features of human disease. Moreover, the virus appears to utilize distinct cellular and molecular pathways in different cell types. Thus, studies of viral infection in human fibroblasts or other skin cells may not directly bear on pathogenesis within human neurons. Recently, VZV has been shown to infect neurons derived from human neural stem cells and human pluripotent stem cells, suggesting that stem-cell based approaches for the study of VZV pathogenesis are likely to hold great promise for the development of new treatments. However, most human neuronal cultures systems derived from stem cells are typically comprised of few, if any, sensory neurons. This is a great limitation since sensory neurons are the main cell type in which the virus establishes latency and later re-emerges during reactivation. Recently, we have developed a human pluripotent stem cell based model of sensory neuron infection by VZV, in which the neurons are capable of harboring VZV in a latent state. We anticipate that this model will allow characterization of cellular and molecular mechanisms of latency and will serve as the basis to develop new treatments for VZV reactivation.

NIH Research Projects · FY 2026 · 2024-01

SUMMARY Allergic diseases represent a major public health burden characterized as an epidemic that involves the interplay between genetic factors and environmental exposures. Allergens are one of the many multiple triggers that produce IgE and cause inflammation of the airways, two crucial components of allergic disorders, including atopic asthma. Spleen associated tyrosine kinase (aka tyrosine-protein kinase SYK) is critically involved in the earliest steps of signaling reaction following aggregation of IgE/FceRI on the basophil/mast cell surface after re-exposure to the same environmental substance (allergen). There is a considerable body of evidence and a consistent set of observations, based on cellular functions and clinical associations, highlighting the deeper relevance of SYK to the IgE-mediated reaction in humans. In addition, we have demonstrated that basophil traits, such as SYK expression and IgE-mediated histamine release, can serve as biomarkers for predicting the clinical efficacy of omalizumab in allergic diseases like asthma. However, little is known about the genetic regulation of SYK, which is crucial in determining how IgE-dependent allergy disorders manifest. We confirmed that the expression levels of SYK and histamine release in human basophils were correlated with single nucleotide polymorphisms (SNPs) in the promoter of the SYK gene. Exploration of cis-expression quantitative trait loci (cis-eQTL) discovered a genomic region encompassing the SYK transcription start site (TSS), and it contained multiple variants with a high potential for transcriptional regulation. We hypothesize that genetic factors interact with specific local cell/tissue environments to modify either SYK expression or its function; therefore defining a unique molecular phenotype of human basophils, which can serve as biomarkers for clinical efficacy of anti-IgE therapy as well as being potential drug targets for novel therapeutics. This application's specific aims are to: (1) determine the genetic regulation of SYK transcriptional activity and function; (2) identify common haplotypes that influence SYK expression and function in individuals of European and African ancestry; and (3) compare transcriptome gene signatures among IgE-bearing cell types. Results from these studies will provide strong motivation to explore the larger impact of their influences on atopic disorders.

NIH Research Projects · FY 2026 · 2024-01

PROJECT SUMMARY/ABSTRACT Anum S. Minhas, MD, MHS, is an Assistant Professor in the Division of Cardiology at the Johns Hopkins University School of Medicine. She seeks a Mentored Patient-Oriented Research Career Development Award in order to obtain essential skills and research experience for an independent career as a physician scientist in women’s cardiovascular health. Her research proposal details a five-year plan, with an overarching goal of understanding the role that pre-pregnancy cardiometabolic risk factors (hypertension, obesity and diabetes) play in preeclampsia disease pathophysiology and associated long-term cardiovascular risk using a lifecourse framework. The specific aims of the research are: 1) to identify distinct subtypes of preeclampsia via a machine learning approach using clinical measures and vascular and inflammatory markers during pregnancy, 2) to examine the impact of pre-pregnancy cardiometabolic risk factors on coronary endothelial function and vascular and inflammatory biomarkers in women with preeclampsia at 3 months postpartum via cardiac MRI, and 3) to determine the association of pre-pregnancy cardiometabolic risk factors and preeclampsia with biomarkers and risk of cardiovascular events at 10 years postpartum. This study will provide novel data that will inform therapeutic strategies to mitigate the risk of preeclampsia itself and of long-term cardiovascular events secondary to its occurrence. Dr. Minhas will receive training in advanced epidemiologic and statistical methods, including machine learning, at the Johns Hopkins Bloomberg School of Public Health. She will master the conduct of clinical, imaging and biomarker studies. Her goals during the career development award period include gaining crucial skills in combining detailed smaller-scale human studies of risk mediators with larger epidemiologic and clinical database studies to help reduce health disparities and lessen the burden of maternal morbidity and mortality. Dr. Minhas has complete support from her mentoring team and institution. Her primary mentor, Dr. Josef Coresh, is an internationally recognized cardiovascular epidemiologist with success training generations of leading clinical researchers. Co-mentors Drs. Allison Hays and Chiadi Ndumele have expertise in novel cardiovascular imaging techniques, women’s health and cardiometabolic disorders. This triad of committed, complementary mentors will help Dr. Minhas achieve her career goals and access the resources and support necessary to transition into an independent academic career as a leader in women’s cardiovascular health research.