Brigham And Women'S Hospital

NIH Research Projects · FY 2026 · 2022-06

PROJECT SUMMARY / ABSTRACT Delivering goal-concordant care is considered a key component of high-value care for seriously ill older adults with multiple chronic conditions (MCC); however, no gold standard for measurement exists. The long- term goal of the proposed work is to advance the science of quality measurement among seriously ill older adults with palliative care needs. The overall objective in this application is to develop a psychometrically valid patient-reported experience measure (PREM) that measures goal-concordant care among seriously ill older adults with MCC. The rationale is that a validated PREM measuring goal-concordant care can provide real- time feedback necessary to deliver patient-centered care that meets the dynamic needs of this population, while simultaneously functioning as a quality indicator for palliative care delivery. The proposed work will complete the overall objective through three specific aims: 1) develop an item pool that measures goal- concordant care based on domains of health-related values among seriously ill older adults with MCC; 2) evaluate measurement properties of the item pool and resultant PREM by conducting Item Response Theory analysis and test-retest reliability of field test data; and 3) determine the acceptability, feasibility, and fidelity of PREM implementation in primary care. The first aim establishes the PREM’s content validity and is a multimethod qualitative study consisting of: 1) semi-structured interviews with seriously ill older adults with MCC and their PCPs and 2) content analysis of serious illness conversation documentation in the electronic health record identified using natural language processing. Themes that emerge from this qualitative process will inform item and scale generation and refine the hierarchy of health-related value domains relevant to goal- concordant care. Items will be further developed and refined via focus groups with patients and geriatric palliative care experts to identify relevant content not already covered; cognitive interviews with patients to format the items; and item review by the study team. The second aim will establish the PREM’s reliability and construct validity and consists of field tests of seriously ill older adults with MCC followed by psychometric analyses to develop and evaluate the items comprising the PREM. Under the third aim, 40 seriously ill older adults with MCC will complete the PREM in a primary care clinic over 48 weeks in a single-arm pilot study with a multistage, mixed methods design to evaluate the acceptability, feasibility, and fidelity of PREM implementation. This proposal is innovative because it will develop the first validated PREM that measures goal-concordant care among seriously ill older adults with MCC that can also be used to measure the effectiveness of advanced care planning. This work is highly significant because this PREM transforms goal- concordant care into a measurable and actionable construct by quantifying the patient’s experience of how effectively care processes support the patient’s ability to attain self-concordant goals. Ultimately, this PREM will help establish a quality standard for palliative care delivery for seriously ill older adults with MCC.

NIH Research Projects · FY 2026 · 2022-05

Project Summary In 2018, nearly 67 million people in the US spoke a language other than English at home and, among them, 40% met criteria for limited English proficiency (LEP). People with LEP are at higher risk of adverse events during medical care. In fact, 1 in 12 patients is at risk for adverse events because of barriers created by language discordance, which occurs when the patient and provider do not share a common spoken language. Surgical disparities have been studied in various disciplines, but very few studies have focused on the impact of language. Understanding the impact that language and language discordance or concordance have on surgical outcomes represents a vital component of providing effective, high-quality patient care. Our overall objective is to improve outcomes for surgical patients with LEP. The central hypothesis is that socioeconomic and sociodemographic factors impact outcomes of surgical care for patients with LEP and that identifying those factors through quantitative and qualitative research rooted in a social ecological model (SEM) will provide an opportunity to design interventions that address these risk factors and reduce surgical disparities. Building on a deeper understanding of patient experiences and social risk factors, this project will test the use of a mobile technology for live medical interpretation provided directly to patients. We seek to determine if such a solution can improve communication and promote patient-initiated language concordant encounters, thereby leading to better surgical outcomes. While some research suggests the effects of language discordance may be mitigated by the use of trained interpreters, qualified interpreters are not reliably used for all patients with LEP, even at resource-rich health centers. These barriers threaten to compromise the provision of patient-centered care for this patient population. To that extent, we will test our central hypothesis by pursing three specific aims: (1) Determine whether language concordance is associated with surgical outcomes in a large retrospective surgical patient cohort; (2) Identify the social factors that influence the use of formal interpreter services to treat primary Spanish-speaking surgical patients with LEP; and (3) Evaluate if an immediate interpreter mobile application enables language concordant communication for primary Spanish-speaking patients with LEP. Likewise, studying the complex interaction of structural, interpersonal, and individual factors will enrich our understanding of the impact of language across the continuum of surgical care. The research proposed in this application is innovative because no one has sought to elucidate precisely how LEP may influence surgical outcomes and nor have they applied the SEM in an attempt to have a profound impact on all populations. Also, by providing the mobile interpretation application directly to patients (in addition to surgeons and nurses), we hope to empower patients with LEP to serve as drivers of their care and improve the patient-centeredness of their care. Ultimately, the findings of this study could be applied to other fields of medicine and patient populations to improve the care of millions of patients by mitigating language and communication barriers to reduce health disparities.

NIH Research Projects · FY 2026 · 2022-05

Project Summary: A major challenge faced by all gene therapies directed to the central nervous system is delivering the therapy across the blood-brain barrier (BBB) in a way that achieves adequate central exposure while minimizing toxicity in non-targeted tissues. The two most common routes of administration for delivering adeno-associated viral (AAV) vectors, the only FDA-approved gene transfer platform for neurological applications, suffer from complimentary limitations. Intravenous (IV) injection can achieve widespread distribution throughout the brain but only at low concentration levels, while intraparenchymal (IPa) injections supply a high concentration of therapy but only near the site of injection. As most neurological disorders manifest in multiple brain regions, successful gene therapy treatments will require an improved approach to AAV delivery that can achieve high therapy concentration and large volume coverage. To address this critical bottleneck, we aim to develop and validate delivery strategies using the technology of focused ultrasound (FUS)-mediated disruption of the BBB to significantly improve the concentration and brain coverage of AAV-packaged gene therapies. In this project we will investigate the application of delivering an AAV-packaged micro-RNA therapy targeting suppression of mutant Huntingtin, the gene implicated in Huntington’s disease (HD). Our preliminary work demonstrates that FUS-BBB opening enhances the delivery of IV-injected AAV1 and AAV9 vectors to targeted brain regions in wild-type and HD model mice. We additionally have experience using our human clinical FUS system to achieve large-volume BBB disruption in rats, monkeys and humans. Building off this preliminary data, Aim 1 will determine the optimal FUS parameters and AAV dose for maximizing AAV concentration at the FUS-targeted site in HD model mice. The safety profile of each treatment will be characterized in terms of markers of neuroinflammation and signs of trauma-like damage to tissue. To maximize brain coverage, we will characterize the safety profile and AAV delivery efficacy as a function of BBB opening volume in HD model mice. Aim 2 will incorporate the information gained in Aim 1 to devise an optimal treatment strategy that balances high AAV concentration, large volume coverage and safety. We will assess the safety and therapeutic efficacy of delivering an AAV-packaged microRNA to HD model mice at 1-, 3- , 6- and 12-month time points in terms of mHTT lowering and functional markers of disease progression. In Aim 3, all of the mice data will be used to devise a treatment strategy for AAV delivery in rhesus macaque monkeys. A safety and biodistribution study will be carried out in these non-human primates using our human clinical FUS system. If successful, the strategies developed here would enable AAV delivery to the brain with a more favorable safety and efficacy profile than existing alternatives and unlock the clinical promise of these transformative therapies.

NIH Research Projects · FY 2025 · 2022-05

Mutations in PKD1 are responsible for over 85% of cases in autosomal dominant polycystic kidney disease (ADPKD). A number of studies by us and others have implicated that the ADPKD proteins polycystin-1 and -2 (PC1 and PC2) modulate a number of cellular events and signaling pathways such as Ca2+ signaling, JAK- STAT, mTOR, cyclic AMP (cAMP), and planar cell polarity (PCP). How polycystins modulate these pathways, however, remains elusive. The sequence of these signaling events is unknown. A major challenge is that many experiments have been performed using different model systems, different cell types, and under different conditions. Better understanding of the cystogenic mechanisms and disease progression of ADPKD is a high priority for clinical care. Our long-term goal of this proposal is to identify and modulate the factors controlling cyst formation and enlargement in ADPKD using multidisciplinary approaches. We previously reported a biochemical interaction between PC1 and protein kinase C and casein kinase substrate in neurons 2 (Pacsin 2), a cytoplasmic phosphoprotein that has been implicated in cytoskeletal organization and vesicle trafficking. By in vitro studies, we found that PC1, Pacsin 2 and N-Wasp are in the same protein complex and deficiency of either PC1 or Pacsin 2 leads to defects in actin cytoskeleton and cell migration in cultured cells. Here we propose to extend our study of Pacsin 2 to multiple orthologous mouse models of ADPKD that we have developed to understand mechanisms suppressing cystogenesis. We aim to develop an in vivo imaging protocol of cell migration and to elucidate the role of Pascin 2 in cystogenesis using multidisciplinary approaches including the analysis the cystogenic proteome and phosphoproteome in human and mouse ADPKD models using the latest quantitative proteomics technology, coupled with innovative bioinformatics tools. The proposed studies will likely discover novel therapeutic targets central to the early event(s) in cystogenesis.

NIH Research Projects · FY 2026 · 2022-04

Delayed arrival to the hospital in stroke is a major unsolved problem in public health that leads to stark and persistent population differences in stroke outcomes. The delay generates health outcome differences because patients from certain demographics arrive later leading to less access to treatment and worse outcomes. The most common reason for delay is the time spent by the patient and witnesses who decide together to watch-and-wait or go to the hospital. Therefore, we propose that social connectedness is a major determinant of the delay phenomenon. Our team has demonstrated that social network structure around a specific patient determines the flow of information that leads to decisions to act rapidly or slowly. Patients who arrived early had large and loosely connected networks, while those who arrived late had small and close-knit networks. What remains lacking, however, is knowledge of the extent of the social network effect in a broader population of stroke patients, its mechanism, and translation into interventions to improve stroke delay and differences in the population. This understanding is critical to establishing rigor and premise for future social network interventions aimed at reducing differences in stroke outcomes. Our long-term goal is to design network-based interventions that reduce delay during stroke and ensure access to therapies. Therefore, in this project, we use a dual empirical and social simulation approach to characterize and model social network effects in a broad patient population. In Aim 1, we will determine whether social networks affect delay in hospital arrival after stroke differentially by race and socioeconomic status. We will capture social network data and time to arrival in 500 patients during their hospital admission. In Aim 2, we will model the potential of network interventions to improve stroke delay. Using data from the same 500 patients and persons in their network, we will parameterize an agent-based model to represent the dynamic decision-making within the social network during stroke. Then we will evaluate the potential effects of network interventions to improve delay and differences within the model. Our central hypothesis is that social network metrics will be associated with hospital arrival time, social networks will moderate race and SES differences in arrival time, and that network interventions such as increasing network size will improve outcomes and differences in social simulations. We have assembled a multidisciplinary team with expertise in stroke, social networks, agent-based modeling, and health differences to execute this project. The proposed research will provide much needed empirical data on social network effects and the potential of network interventions to address stroke delay and its differences in the population. These results will have a positive impact by directly setting the stage for testing social network interventions in acute stroke clinical trials to improve arrival time and enhance access to stroke therapies.

NIH Research Projects · FY 2026 · 2022-04

PROJECT SUMMARY / ABSTRACT Rheumatoid arthritis (RA) is a systemic inflammatory disorder affecting approximately 1.5 million people in the United States. Although parenchymal lung disease, which encompasses interstitial lung disease (ILD) and emphysema, is a common lung complication of RA with increasing prevalence and mortality, no strategies currently exist for early detection or risk stratification of progressive disease. This is an area of unmet need that could lead to improved opportunities for intervention and a decrease in the considerable morbidity and mortality of RA-associated parenchymal lung disease. In this proposal, we aim to fully characterize early parenchymal lung disease in RA and establish risk factors for progressive disease. We hypothesize that clinical and molecular risk factors can predict the development and progression of RA-associated parenchymal lung disease, including preclinical ILD (preILD) and emphysema, and that the molecular profile of progressive RA-preILD will overlap with fibrotic RA-ILD. To test this hypothesis, we will propose the following: In Specific Aim 1, we will determine the prevalence and progression of RA-associated preILD and emphysema using visual and objective radiologic approaches and correlate these measurements with functional capacity, respiratory symptoms, and health-related quality of life assessments. In Specific Aim 2, we will define clinical and molecular determinants that predict the development and progression of RA emphysema and compare the molecular profile of emphysema with preILD to provide mechanistic insight into the divergent patterns of parenchymal lung injury caused by RA. In Specific Aim 3, we will demonstrate that progressive RA-preILD and fibrotic RA-ILD have similar molecular signatures, suggesting phenotypic and mechanistic overlap. To achieve the aims of this proposal, a longitudinal cohort of 200 RA patients without a history of ILD will be followed for up to 6 years with detailed clinical, functional, radiologic, and molecular phenotyping. The successful completion of this research will provide us with a better understanding of the early characteristics and natural history of RA-associated ILD and emphysema and establish novel non-invasive ways to identify those at risk for progressive disease. This will enable closer monitoring and earlier opportunities for intervention, potentially leading to decreased morbidity and mortality in individuals afflicted with RA-associated parenchymal lung disease.

NIH Research Projects · FY 2025 · 2022-04

PROJECT SUMMARY Pathology driven by resident cells in the central nervous system (CNS) such as astrocytes and microglia plays an important role in multiple sclerosis (MS), particularly for its progressive stage which is eventually achieved by most MS patients. However, no drugs are available for the modulation astrocyte and microglia pathogenic activities. We developed an in vitro assay to identify compounds that modulate astrocyte pathogenic activities. These studies identified compound A-38, an inhibitor of the Erythropoietin-producing hepatocellular carcinoma receptor B3 (EphB3), as a suppressor of astrocyte pathogenic activities and, consequently, as a lead therapeutic compound for progressive MS (PMS). Moreover, our data suggest that microglia-driven EphB3 signaling in astrocytes promotes CNS inflammation and neurodegeneration. Ephrin receptors are known to participate in CNS development, however our findings define a novel role for EphB3 in CNS inflammation and identify it as potential target for therapeutic intervention. We hypothesize that EphB3 kinase inhibition will provide an efficacious therapeutic approach for PMS, and potentially other neurologic diseases. Thus, we propose to develop new A-38 analogs to treat PMS. Our Specific Aims are: SPECIFIC AIM 1: OPTIMIZATION AND IN VITRO EVALUATION OF EPHB3 KINASE INHIBITORS. We will design, synthesize and evaluate the properties of up to 150 analogs of A-38. The goal of these studies is to optimize the potency and kinase selectivity of new analogs of A-38, while maintaining good drug-like properties and brain exposure following oral delivery. During the R61 phase of this project, we will integrate iterative structure-activity relationship (SAR) studies and structure based drug design to optimize A-38 potency and kinase selectivity, while maintaining good drug-like properties. We will first evaluate A-38 analogs for EphB3 activity and for selectivity in EphB2 EphB4, and EphA4 kinase assays. Promising compounds will advance to phenotypic assays to identify those molecules that modulate astrocyte pathogenic activities. Compounds of interest will then be characterized in drug-like property assays to select leading compounds for PO PK studies. At the end of the R61 phase of this project we expect to identify a lead EphB3 inhibitor and 1-2 back-ups suitable for advancement into in vivo efficacy testing in the R33 phase. SPECIFIC AIM 2: EVALUATION OF LEAD COMPOUND(S) IN PMS PRE-CLINICAL MODELS. During the R33 phase of this project, we will evaluate the lead EphB3 inhibitor and 1-2 back-ups in the NOD EAE and cuprizone-induced murine pre-clinical models which recapitulate several aspects of PMS. We will evaluate the effects of the compounds on disease development, CNS inflammation, axonal loss and demyelination, as determined by histopathology, flow cytometry and gene expression analyses. In summary, this project will identify a novel EphB3 inhibitor with drug-like properties suitable for development as a candidate therapeutic for PMS and other neurologic diseases with NIH and/or industry support.

NIH Research Projects · FY 2025 · 2022-04

Project Summary/Abstract Protein misfolding is a key feature of many human diseases including most neurodegenerative diseases and many cancers. Destruction of misfolded proteins is largely mediated by the proteasome, a 2.5 MDa multisubunit complex which is the most sophisticated protease ever described. The proteasome's active sites are sequestered within a barrel-shaped cylindrical chamber, known as the core particle (CP). Access of substrates to the CP is mediated by the regulatory particle (RP), which recognizes proteasome substrates via their ubiquitin tags. The RP unfolds, deubiquitinates, and injects the substrate into the CP where it is rendered into small peptides. Pharmacologic inhibition of the proteasome is an established anti-cancer therapy, most notably in multiple myeloma. Conversely, the possibility of enhancing proteasome function has generated considerable interest in recent years. Such a strategy might ameliorate diseases caused by protein misfolding. A key step in the generation of active proteasomes is the assembly of the 700 kDa 28-subunit CP, which precedes assembly of the full proteasome and occurs by an ordered multistep pathway that requires the function of five dedicated chaperone proteins. Structural analysis of CP maturation has been hampered by challenges in isolating and characterizing assembly intermediates due to their low abundance and transitory nature. Here we hypothesized that defined CP mutants may be enriched for assembly intermediates. We have developed a productive work-flow for the affinity purification and structural analysis of these mutants, and have already generated eight high resolution structures. In Aim 1, we will carry out this structural analysis of CP mutants using Cryo-Electron Microscopy, coupled with detailed structure-function analyses. In Aim 2, we will characterize a long-known but poorly understood regulator of the CP known as PI31/Fub1. We will attempt to determine its structure in complex with the CP using Cryo-Electron Microscopy, and test a number of specific hypotheses regarding its function. In Aim 3, we will characterize a novel protein which is a previously unrecognized transcriptional target of the Rpn4-mediated proteasome biogenesis regulon, and which appears to be a new proteasome-interacting protein. This proposal is expected to provide significant insight into proteasome assembly and overall function, information which could lead to novel therapeutic strategies based on modulating proteasome activity to treat diseases characterized by protein misfolding.

NIH Research Projects · FY 2025 · 2022-04

Project Summary Elastic fibers provide the elasticity that is essential for the function of many organs/tissues, such as lung, large arteries, and skin. Congenital deficiency or destruction/degradation of elastic fibers can lead to emphysema, arterial aneurysm, and laxity of tissues. Reversely, excessive formation of elastic fibers is a feature of pleuroparenchymal fibroelastosis, which is currently without any effective treatment. Many proteins are modified after they are produced in cells. Such modification can critically influence the function of proteins. One such modification is citrullination, which has been shown to regulate the function of various types of cells. Aberrant citrullination has been observed in many human diseases, such as chronic obstructive lung disease, idiopathic pulmonary fibrosis, and rheumatoid arthritis. Thus, manipulating protein citrullination can be beneficial in many clinical settings. This project is based on a novel observation that citrullination critically regulates the formation of elastic fibers. Its goal is to investigate the role and mechanism of action of citrullination in regulating the formation of elastic fibers and lung function. Molecular, cellular, and biophysical approaches will be deployed to elucidate how citrullination regulates the elastogenesis of fibroblasts (Aim 1) and how citrullination of critical elastogenic proteins is regulated (Aim 2). Reagents detecting the citrullination of the critical elastogenic proteins will be developed to facilitate this endeavor. Finally, genetically engineered mice will be generated to examine cell type-specific and temporal roles of citrullination in modulating the function of lungs and the development of emphysema induced by cigarette smoking (Aim 3). Knowledge gained from this project very likely will uncover novel therapeutic approaches toward many human diseases, such as emphysema and arterial aneurysm.

NIH Research Projects · FY 2026 · 2022-04

PROJECT SUMMARY/ABSTRACT Heart failure with preserved ejection fraction (HFpEF) is a growing epidemic associated with multiple comorbidities, including hypertension. In part because of its heterogeneity, clinical trials in HFpEF have been largely disappointing to date, though the NHLBI-funded TOPCAT trial of spironolactone, a mineralocorticoid receptor (MR) antagonist, had promising results. This has prompted several ongoing NIH and industry trials of MR antagonists in HFpEF. However, the mechanisms through which MR antagonists may treat HFpEF remain poorly understood. The MR is expressed in cardiomyocytes and vascular endothelial and smooth muscle cells. Activation of the MR at these sites causes inflammation, oxidative stress, endothelial dysfunction, and fibrosis – all pathways implicated in HFpEF. Primary aldosteronism (PA) is a cause of both hypertension and disproportionate cardiovascular risk that is treated with MR antagonists, which target the underlying pathophysiology: renin-independent production of aldosterone that chronically activates the MR. Though considered rare, PA has recently been identified in up to 15-20% of apparently “essential” hypertension, with a continuum of severity ranging from subclinical to overt, paralleling blood pressure and responsiveness to MR antagonists: termed ‘subclinical PA.’ These data suggest an expanded role for MR antagonists as precision therapy in the appropriate patients. Given its prevalence in hypertension, subclinical PA may be an unrecognized mechanism of HFpEF pathogenesis and identify a patient subgroup best treated with MR antagonists. The current proposal aims to evaluate the relationship between subclinical PA and HFpEF risk and to probe the mechanism of risk using physiologic imaging techniques. Specific Aim 1 is to evaluate both the cross-sectional relationship with cardiac structure and function and the longitudinal relationship between subclinical PA and incident HFpEF in the NHLBI Atherosclerosis Risk in Communities (ARIC) cohort. Specific Aim 2 leverages the mentors’ established Adrenal and Cardiac PET registries to investigate the relationship between PA and myocardial flow reserve, a marker of vascular dysfunction and cardiovascular risk, by evaluating the effect of an MR antagonist. This research will be accomplished within a comprehensive career development plan designed to provide Dr. Brown with the skills to become an independent physician-scientist. Her long-term career goal is be an independent, R01-funded physician-scientist focused on understanding the underlying biological and hormonal mechanisms of heart failure risk, using physiologic imaging tools to identify and characterize subclinical phenotypes that can be leveraged to enrich for responsive patient populations in clinical trials, and ultimately to identify targets for heart failure prevention. An outstanding mentoring team and advisory committee of established scientists in the fields of aldosterone and vascular biology, HFpEF, state-of- the-art cardiovascular imaging, cardiovascular clinical trials, and epidemiological and biostatistical methods will guide the candidate’s transition to scientific independence over the course of the award period.

NIH Research Projects · FY 2026 · 2022-04

PROJECT SUMMARY AND ABSTRACT Parkinson's disease (PD) is the most common neurological disease associated with movement abnormality. It has been 25 years since the first genetic cause of PD was identified, and yet there is still no effective treatment for the disease. One of the hinders we think is the lack of models that assess early PD pathogenesis and therapy responses in its real neurophysiological environment. This provides a significant bottleneck in our ability to make progress in this disease. Two lines of recent evidence motivate us to study PD pathogenesis in a real neurophysiological environment: (1) Human neuroimaging data and animal models both showed that synaptic disruption proceeds neuronal death, making the case that PD is a synaptopathy. (2) Many novel, regulatory, non-coding RNAs show linkage to PD pathogenesis. For instance, we found over 20,000 enhancer RNAs (or eRNAs) candidates in dopamine neurons of human post-mortem brains (Dong et al. Nature Neuroscience, 2018). They significantly co-localized with PD risk variants. The other class of novel RNAs is circular RNAs (circRNAs), which are predominantly enriched in the brain, highly specific to the synapse, and ultra-stable (e.g., 10x longer half-life than linear RNAs). We identified >11,000 circRNAs actively expressed in the dopamine neurons, many of which are significantly associated with PD pathology (Dong et al. in submission). More importantly, circRNAs can form a regulatory network with lncRNAs and miRNAs, and can be wrapped into extracellular vesicles (EV), penetrating blood-brain barriers. Based on these, we hypothesize that regulatory RNAs incl. circRNAs, eRNAs, miRNAs, lncRNAs can be detected in EV and might play a role in the synaptic dysfunction in PD pathogenesis. To test this hypothesis, we need a model to recapitulate the dynamic physiological microenvironment of PD pathogenesis. In this study, we will combine our expertise in brain organoids, PD biology, exosome analysis, single-cell omics, bioinformatics, and biomedical engineering to develop a new 3D brain organoids microphysiological analysis platform (MAP) to recapitulate the dopamine neurons' interconnectivity and study molecular neurodegeneration systematically. We will (1) first develop PD organoids and profile the transcriptome (incl. circRNAs, miRNAs, mRNAs, lncRNAs, etc.) of secreted EV and single-cell transcriptome of brain organoids, to identify PD-associated RNAs, then (2) map the pathophysiological dynamics of PD organoids in a novel, high-throughput, mini-brain-on-chip platform, and last will (3) integrate the EV-organoid temporal multi-dimensional data to infer the PD-associated RNAs and their regulatory dynamics during the PD pathogenesis. Recent breakthroughs in RNA therapeutics have led to multiple first-in-human trials and clinical approval (e.g., Moderna, Alnylam, and Ionis pharmaceuticals). circRNAs have many advantages over linear RNAs, making them potentially better suited for translation into therapeutics and diagnostics. EVs secreted from PD organoids provide a good proxy of fluid biopsy for studying PD brain's neuropathology. Thus, this interdisciplinary (neurology, biomedical engineering, computational genomics) study will set an important, highly innovative foundation for understanding PD neuropathology and exosome treatment.

NIH Research Projects · FY 2025 · 2022-04

PROJECT SUMMARY/ABSTRACT The United States does not have sufficient science to mount an evidence-based response to combat the health detriments of present-day anti-Asian American (AA) hate. AA parents and children are struggling with a deeply troubling new sociopolitical context, a context with unprecedented levels of anti-Asian hate and violence fueled by references to COVID-19 as the “China virus.” Physical assaults against AAs skyrocketed by 145% in 2020 and have spiked to 164% in the first quarter of 2021. Among AA youth, 80% report being bullied or verbally harassed and 24% reported being shunned. Our evidence base for how racism impacts AA mental health under “normal” circumstances is insufficient and woefully inadequate. This R01 fills a critical gap in the science of how discrimination affects AA adolescent mental health, how adolescents respond to racial discrimination, and the agents involved in racial socialization processes involving AA adolescents. There are no longitudinal studies that investigate the effects of racial discrimination on AA adolescent mental health while also incorporating novel observational methods to determine how parents, peers, and social media are racial socializing AA adolescents. This project will follow 350 Chinese American adolescents (12-15 years), their Chinese heritage parent, and a peer to investigate the effects of discrimination experiences, discrimination responses, and racial socialization processes on adolescent mental health (depression, anxiety, suicidality) and on chronic stress (hair cortisol) over three timepoints. The long-term objective is to develop a biomedical evidence base focused on how parents, peers and social media can be leveraged to mitigate the negative health consequences of discrimination. Our multi-method approach (observation, survey, interview, biomarker) investigates the following aims: Aim 1. To assess how racial discrimination is associated with Chinese American adolescent (a) mental health (depression, anxiety, suicidality) and (b) biomarkers of chronic stress (hair cortisol) over three years. Aim 2. To determine how adolescent characteristics (internalization of the model minority myth, ethnic identity) moderate the association between discrimination and mental health/chronic stress. Aim 3. To (a) understand how parent characteristics (racial socialization, mental health, ethnic identity), moderate the association between adolescent discrimination and adolescent mental health/chronic stress (hair cortisol). We also consider “upwards socialization”; how adolescents are active agents in their own socialization and the socialization of those around them. Discrimination during adolescence has especially pernicious effects on concurrent and downstream adult health and understanding AA adolescent discrimination will help identify those at risk for problematic mental health. This study will inform urgently needed individual-, family- and peer-based interventions to address mental health for AA adolescents who are facing unprecedented risks to mental health at this time.

NIH Research Projects · FY 2026 · 2022-04

PROJECT SUMMARY Tuberculosis (TB) is the leading cause of death among children with HIV, yet insufficient data are available on the pharmacokinetics (PK) of newer HIV/TB cotreatment and TB prevention strategies in children. Global expansion of dolutegravir use for young children has the potential to significantly improve HIV treatment outcomes, but there are no PK data to inform use of double-dose dolutegravir during rifampicin-containing HIV/TB cotreatment for children under 6 years of age. Further, multiple studies have shown that current WHO-recommended rifampicin dosages result in low concentrations in most children, and high-dose rifampicin may improve outcomes and shorten treatment duration in adults and children. Yet the impact of high-dose rifampicin on dolutegravir exposures has not been examined. Finally, it is well-established that treatment of latent TB infection (LTBI) significantly reduces incident TB among HIV-infected persons, yet this strategy is vastly underused in endemic settings. Short-course LTBI treatment regimens increase completion rates, but have not been studied among HIV-infected children receiving dolutegravir. To address these gaps in knowledge and provide needed PK and safety data to extend use of these promising strategies from adults to children, we will carry out two prospective PK studies to examine: (1) twice daily dolutegravir during both standard- and high-dose rifampicin, and (2) PK of dolutegravir during weekly rifapentine/isoniazid for TB prevention/LTBI treatment. To advance our understanding of underlying mechanisms of drug action we will also examine the impact of standard- and high-dose daily rifampicin and weekly rifapentine on the 4β-hydroxy-cholesterol to cholesterol ratio, an endogenous biomarker of CYP3A4 activity. This will provide critical pharmacodynamic (PD) data for use in population PK/PD models to quantify variability and inform optimal drug dosing in this vulnerable population of children. Nigeria is home to more children living with HIV than any other country in the world, and also bears one of the greatest burdens of incident TB. Since 2004, APIN Public Health Initiatives has provided HIV care and treatment to over 20,000 children in Nigeria and thus is uniquely positioned to support this vital PK research. This combination of studies addresses critical gaps in knowledge regarding PK and safety of newer drug combinations for HIV/TB cotreatment and TB prevention, findings that may inform WHO guidance on use in children, toward the goal of curbing TB’s devastating toll in this vulnerable population.

NIH Research Projects · FY 2025 · 2022-04

Summary It is well known that sleep loss is associated with significant short- and long-term health consequences, but to date the impact of sex differences in response to sleep loss are poorly understood. We and others have shown that women have greater neurobehavioral performance impairment than men when exposed to one night of acute sleep loss. Furthermore, when this effect is examined separately by menstrual cycle phase, women in the follicular phase exhibit greater impairment compared to both men and women during the luteal phase, suggesting a possible endocrine mechanism. Indeed, we have shown that these differences in performance may be driven by sex-steroid-mediated changes in core body temperature (CBT), specifically involving the ratio of progesterone (P4) to estradiol (E2) between the follicular and luteal phases. These findings have important implications for understanding the interactions of sleep loss and female sex hormones on neurobehavioral performance and other health consequences. An important open question, however, is how these menstrual-phase-dependent differences impact performance under more realistic patterns of chronic sleep loss. Millions of women routinely obtain less than the recommended 7-9 hours of sleep per night during the week and attempt to catch up on sleep on the weekend. In men, we have shown that any apparent improvement during such recovery sleep is transient, and that subsequent sleep loss results in more accelerated deterioration in performance. It is unknown how this variable pattern of chronic sleep loss and recovery sleep impacts performance in women during the follicular and luteal phases of the menstrual cycle. The proposed work will fill this important gap in knowledge. In our proposed 11-day inpatient study, healthy premenopausal women will be randomized to either chronic variable sleep deficiency (with a repeated pattern of two nights of 3 hours time-in-bed followed by one night of 10 hours time- in-bed, equivalent to our prior study in mean) or a sleep satiation control (10 hours time-in-bed throughout) during either the follicular or luteal phase of their menstrual cycle. This protocol will allow us to quantify both the impact of chronic variable sleep deficiency on neurobehavioral performance in women and differences in the response to chronic variable sleep loss across the menstrual cycle. We will also investigate the role of CBT, P4, and E2 in mediating these responses. As an exploratory analysis, we will also evaluate the impact of chronic variable sleep loss on E2 and P4 levels to investigate the effect of insufficient sleep on circulating levels of female sex hormones across the menstrual cycle. There is immediate

NIH Research Projects · FY 2026 · 2022-04

PROJECT SUMMARY / ABSTRACT Traumatic brain injury (TBI) is a major health problem; 2.5 million Americans sustain TBI each year at a cost of 80 billion dollars annually. Few therapies reduce long-term cognitive sequelae of TBI, and there are only limited options for rehabilitation. Brain injury causes a primary structural injury followed by a secondary phase, which involves the activation of the innate and adaptive immune systems. Neuroinflammation with microglia’s involvement has been identified as a major contributor to the pathogenesis of TBI. However, there is still no effective immune therapy to modulate the microglial response post-injury. Nasal administration of anti-CD3 monoclonal antibody induces an anti-inflammatory immune response that down-regulates microglial activation in animal models of multiple sclerosis. The mechanism involves localization of nasal anti-CD3 to cervical lymph nodes where it induces IL-10-secreting (CD4+LAP+ and CD4+FoxP3+) regulatory T cells (Tregs) that migrate to the brain and inhibit microglial activation. Nasal anti-CD3 therapy is an unexplored area in TBI, and the mechanisms by which Tregs modulate microglia in TBI are largely unknown. Dr. Saef Izzy, a trained neurointensivist with a background in acute brain injury research, hypothesized that nasal anti-CD3 represents a unique, clinically applicable immunomodulatory approach for the treatment of TBI. Dr. Izzy worked closely with his primary mentor, Dr. Howard Weiner, to investigate the effects of nasal anti-CD3 on TBI outcomes in mice. His preliminary data showed that nasal anti-CD3 increases CD4+Tregs and IL 10 expression and reduces microglial activation in the brain 7 days after controlled cortical impact injury (CCI). It also improves behavioral outcomes at 1-month post-injury. In this K08 proposal, Dr. Izzy will determine the effects of nasal anti-CD3 on long-term histopathological and behavioral outcomes after CCI (Aim 1). He will survey the microglial, effector, and regulatory T cell responses after injury and study the effects of Tregs on microglial inflammatory response in vitro after CCI (Aim 2). He will investigate the effect of nasal anti-CD3 on IL-10/IL- 10R signaling in microglia using a C57BL6/J mouse harboring IL-10Rflox/floxTMEM119CreETR2, which does not express the IL-10 receptor on microglia after tamoxifen administration. He will also delineate the role(s) of other anti-inflammatory cytokines produced by Tregs by neutralizing TGF-β and IL-4 in vitro and in vivo and study their effects on behavior and microglial inflammatory response post-CCI (Aim 3). Dr. Izzy's career goal is to better understand how the adaptive immune response interfaces with the innate immune system after TBI. Successful completion of this proposal will provide insight into the mechanisms by which Tregs modulate the microglial response after TBI and the identification of novel immune-based therapeutics to improve patients’ outcomes. The proposed K08 application leverages Dr. Izzy’s mentor’s expertise in immunology, mouse models of acute brain injury, and bioinformatics to provide him with the additional knowledge and experience necessary to become an independent NIH-funded investigator and expert in the neuroimmunology of TBI.

NIH Research Projects · FY 2025 · 2022-04

Project Summary/Abstract With age, humans can accumulate leukocyte clones in blood that arise from somatic mutations in bone marrow stem cells that enhance expansion: clonal hematopoiesis of indeterminate potential (CHIP). Mutations in DNMT3A and TET2 account for the plurality of these clones. CHIP confers highly elevated cardiovascular (CV) risk, independent of traditional risk factors. We have found accelerated atherogenesis and the involvement of IL-1 and IL-6 in mice with myeloid loss of Tet2 or Dnmt3a function and that genetically reduced IL-6 signaling abrogates the elevated CV risk in humans with DNMT3A or TET2 CHIP. Analyses of our CANTOS trial showed greater efficacy of IL-1β inhibition in humans with DNMT3A or TET2 CHIP. These results point the way to a genotype-directed allocation of therapy, an approach that has transformed oncology but remains aspirational in atherosclerosis. The roles of classical vs. trans IL-6 signaling in atherothrombosis requires further study due to conflicting evidence. Specific aim 1 will test the hypothesis that atherosclerotic mice that mimic CHIP due to myeloid deficiency in Dnmt3a treated with an antibody that interrupts global IL-6 signaling by neutralizing IL-6 receptor α (IL-6r, CD126) have decreased atherosclerosis and inflammation within the lesions, blood and other organs as gauged in part by single-cell RNA sequencing (scRNA-seq). We further hypothesize that IL-6r inhibition will limit expansion of the mutant clone. Specific aim 2 will probe the role of classical vs. trans IL-6 signaling in myeloid cells in accelerated atherogenesis in CHIP using Il6rflox/flox/Lyz2-Cre mice bone marrow chimeric LDLR-/- mice (to block leukocyte classical signaling), and administration of a gp130-IgG1-Fc chimeric protein (to block trans signaling) using similar procedures and endpoints. The results will illuminate the controversy and unsettled science regarding the contributions of classical and trans IL-6 signaling to atherogenesis. Specific aim 3. Our preliminary experiments show that female myeloidTet2-/-Ldlr-/- mice have greater acceleration of atherogenesis than Tet2+/+ Ldlr-/- male mice and, unlike their male counterparts, show reduced atherogenesis with IL-1β neutralization. We will localize where this sex difference operates in the inflammasome–IL-1β–IL-6 pathway, and test the hypothesis that female Dnmt3a-/-Ldlr-/- mice have greater response to IL-6r inhibition than males. We will probe mechanisms by gonadal ablation experiments and by analysis of scRNA-seq data, which in preliminary data shows IL-1β expression in resident macrophages cells from atheroma from Tet2-/- females but not males. This work will deepen understanding of the mechanisms of accelerated atherosclerosis in CHIP. Our new pilot clinical data show that an anti-IL-6 antibody can mute inflammation in humans. Thus, the work proposed here will provide an indispensable step toward validating and furnishing the fundamental basis of an immediately translatable targeted therapeutic strategy based on CHIP genotype.

NIH Research Projects · FY 2026 · 2022-04

PROJECT SUMMARY/ABSTRACT Chronic obstructive pulmonary disease (COPD) is a leading cause of respiratory mortality worldwide15. Identifying highly susceptible individuals early in their disease course and understanding pathogenic mechanisms, before irreversible loss of lung function, is of utmost importance16,17. Genetics account for about 40% of COPD susceptibility18–20. Genome-wide association studies (GWASs) have identified multiple variants associated with COPD21–23. Individual variants are poor for risk prediction, but in aggregate genetic variants can account for a substantial portion of risk. Pooling millions of GWAS variants, I created a polygenic risk score (PRS) for COPD that can identify individuals at high risk for COPD, though performance was less optimal in non- Europeans24. Multi-ancestry PRSs are needed as genetic ancestry is not readily determined in clinical practice. Further, gene expression, reflecting genetic and environmental influences, provides pathobiologic information for COPD susceptibility and heterogeneity. A transcriptional risk score (TRS) for COPD that adds predictive value above clinical risk factors25 has yet to be developed. The appeal of using -Omics data for risk stratification is that these data can lend insight into why certain COPD subgroups are at elevated risk of progression. Gene regulatory networks26 have been used to uncover mechanisms of COPD heterogeneity that were not found by traditional gene-based approaches. Therefore, we hypothesize that polygenic and transcriptional risk scores will substantially improve upon clinical factors in identifying those at higher risk for COPD and related phenotypes, and can be used to identify pathways for therapeutic intervention. We will train multi-ancestry PRSs using 4,225 African ancestry individuals from UK Biobank and existing analyses of 8,429 African-Americans from CHARGE, and test in the Genetic Epidemiology of COPD (COPDGene: n=10,198) study and Lung Tissue Research Consortium (LTRC: n=1,078). We will create a multi-ancestry transcriptional risk score (TRS) using whole blood RNA-sequencing (RNA-seq) data in training (n=3,394) and evaluate predictive performance in testing samples (n=1,131) of COPDGene. We will use Connectivity Map (CMap)8,27 to identify drug repurposing candidates based on TRS transcripts. We will leverage lung RNA-seq data from LTRC to create a lung TRS, and test in COPDGene blood samples. We will classify COPDGene participants along the axes of the existing PRS and lung TRS (e.g. “High” PRS, “Low” TRS), which we expect will identify those at high risk for COPD-related phenotypes and progression. To understand why certain individuals are at high risk for COPD phenotypes, we will utilize gene regulatory networks to identify pathways differing between PRS/TRS classifications, and use the Gene RegulAtory Network Database (GRAND)9 to prioritize drug repurposing candidates. These aims will generate data for future studies, which will focus on validating COPD -Omics risk scores and drug candidates in real-world cohorts1, and using machine learning to predict the network effects of drug candidates. The proposed research and career development plan will train me to use machine learning for multi-omic integration and risk prediction.

NIH Research Projects · FY 2026 · 2022-03

To better understand the molecular and cellular pathways driving autoimmune diseases, we propose to deeply profile biological samples from patients with rheumatoid arthritis (RA), psoriasis (Ps), psoriatic arthritis (PsA), primary Sjögren’s syndrome (pSS) and systemic lupus erythematosus (SLE). Working closely with the AMP AIM network, we will focus on the cohorts and clinical questions defined by the network of clinicians, biologists and computational biologists together with industry and non-profit partners. For our Technology Core, we selected leading-edge multi-dimensional technologies to deeply profile end organs as well as peripheral blood from patients with autoimmunity. We assembled a team of investigators with expertise in each disease and tissue type, and who have already demonstrated the ability to develop and implement high- throughput pipelines to profile tissue and blood samples. To help us design and interpret the studies, we recruited a team of collaborators and consultants with clinical expertise in each disease, with pathology expertise in each tissue and with technical expertise in spatial profiling methods. In the first year, we will carry out the pilot phase to optimize pipelines for: i) preserving and disaggregating tissues; ii) profiling single cells using scRNA-seq/CITE-seq/TCR-BCR-seq; iii) profiling single nuclei by snRNA-seq/snATAC-seq; iv) preparing tissues for two complementary, leading-edge methods for spatial transcriptomics (Visium) and transcript imaging (MERFISH). We will iterate the protocols to optimize cell viability and yield, technical quality metrics specific to each technology and the representation of all cell types. In year 2, we will run the full set of scaled- up technology pipelines using ~50 samples per tissue type, and will assess data quality, site and batch effects and technical artifacts to inform potential modifications for the single-cell pipeline. In Years 3-5, we will profile the remaining ~1000 biopsies and ~1000 blood samples collected by the disease teams. We will develop a computational pipeline for data pre-processing, primary biological analysis and quality metrics (including technical and biological parameters at the scale of genes, cells and tissues) with customized features for each tissue and disease. Our findings will be rapidly communicated within our Technology Core and across the Network to the Disease Teams, Systems Biology Core, network committees overseeing the project, NIH, FNIH and our industry and non-profit partners. We will also respond in real time to advancing changes in technologies and work with the network to pilot and scale up critical methods. The result of the proposed studies will be a set of multi-dimensional datasets that will be shared with the network and the larger community, and provide a basis for cutting edge disease deconstruction and reconstruction across autoimmune end organ pathologies and thus fulfill the vision of AMP-AIM.

NIH Research Projects · FY 2026 · 2022-03

PROJECT SUMMARY/ABSTRACT Here we propose a Systems Biology Core (SBC) for the Accelerating Medicines Partnerships in Autoimmune and Immune-Mediated Diseases (AMP AIM). The AMP AIM will use high dimensional molecular and cellular assays to define the key cell states, pathways, and molecular components of tissue inflammation and damage by examining patient tissue and blood samples. Ultimately, we seek to define the components of tissue inflammation in autoimmune and inflammatory diseases including psoriatic spectrum diseases (PSD), rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), Sjogren’s syndrome (SS), and other related conditions. AMP RA/SLE initiated this process by querying 106 single cells in inflamed RA synovial and SLE nephritis tissue samples using multimodal strategies; it defined key cell states in tissue inflammation, including T peripheral helper T cells (Tph), GZMK+ CD8+ T cells, HLA-DR+THY1+ fibroblasts, and autoimmune- associated B cells (ABCs). Now, to understand how these and emerging cell-states function and interact to cause disease, it will be essential to obtain high dimensional data on patient sample data across a spectrum of diseases and disease sub-phenotypes. These data may capture the cellular states; the spatial localization of cell states, proteins and transcripts; histological features; and other tissue parameters. A powerful and skilled team that is able to define strategies to analyze this data, integrate multiple modalities of data, and integrate results from across a diverse set of diseases and tissues will be essential to the success of this program. We build from our experience leading the Systems Biology Group within the Accelerating Medicines Partnerships Rheumatoid Arthritis and Systemic Lupus Erythematosus (AMP RA/SLE). We have built a team that is skilled at analysis of diverse modalities and computational biology. We have specific experience and expertise in inflammatory diseases. Here we propose to: (1) Develop Tools and Technology to analyze high dimensional cellular and molecular data. This includes optimizing existing bioinformatics and computational tools. It also includes developing new computational and statistical methods to integrate high dimensional data manifestation of disease. (2) Enable collaboration throughout the network and facilitate systems level analysis. We envision that this is an integrated activity with the network, where we will devise and ultimately create an integrated model of tissue inflammation across diseases to define features that drive clinical disease. This will require the development of new statistical and computational methods. It will also require tight collaboration within the network including data synchronization, storage, sharing, and clinical data integration. In addition, we will engage the network by offering consultation, technical support and training in high dimensional data analysis. .

NIH Research Projects · FY 2025 · 2022-03

Program Director/Principal Investigator (Last, First, Middle): Crosby, Gregory J. ABSTRACT Delirium is the most common complication of surgery and anesthesia in older patients, afflicting 15-60% of those having non-cardiac procedures. This is a major clinical problem because nearly 40% of surgical procedures are performed on older patients and delirium is associated with serious morbidity, including accelerated decline to dementia. Still, the cause of postoperative delirium is uncertain and its neuropathogenesis is unknown. Cerebral neurodegenerative pathology characteristic of Alzheimer’s disease and related disorders is common even in cognitively normal older people, and by the time even mild symptoms develop is pronounced. We hypothesize that this preexisting burden of occult cerebral neurodegeneration accounts for enhanced vulnerability to postoperative delirium (Aim 1), augments surgery- and delirium-induced cerebral injury (Aim 2), and fuels surgery- and delirium-induced inflammation, which aggravates neural injury and increases neurodegeneration (Aim 3). We will test these hypotheses in a longitudinal prospective observational study of older surgical patients evaluated preoperatively for cognitive impairment and followed postoperatively for delirium and longer-term cognitive decline, with neurodegeneration and neural injury defined by ultrasensitive plasma biomarkers and cerebral MR imaging. This project is innovative because it addresses the key question of whether surgery and anesthesia produce delirium and persistent cognitive decline de novo or by unmasking or exacerbating a cerebral precondition and is significant because of the magnitude of the clinical problems (neurodegenerative disease, delirium), practicality of the approach (blood- based biomarkers), and potential to improve the outcomes of vulnerable older surgical patients. OMB No. 0925-0001/0002 (Rev. 03/2020 Approved Through 02/28/2023) Page Continuation Format Page

NIH Research Projects · FY 2026 · 2022-03

Project Summary Neurodegenerative diseases, especially Alzheimer’s Disease (AD) and Parkinson’s Disease (PD), affect millions of people globally and represent a US major healthcare burden. AD is the world’s most common cause of dementia, and is the sixth leading cause of death in the US. PD is motor disabling condition that advances to cognitive deficits. Currently there exist no effective therapies for AD or PD; even some of the drugs used to ameliorate symptoms, such as in PD, cause serious long-term side effects that may be worse than the disease itself. In AD and PD, accumulation of Amyloid-β (Aβ) or α-Synuclein (α-Syn) and their subsequent aggregation, cause neuronal toxicity, including neuroinflammation, synaptic deficits and neurodegeneration, leading to cognitive or motor deficits. In this application, we conceive an innovative genetic tool (named Autophagon, or AFN) to be developed as a gene therapy that helps target toxic Aβ or α-Syn species to autophagy, an important degradative pathway that is usually impaired in AD and PD. AFN will feature a synthetic gene fragment to help sequester Aβ or α-Syn aggregates and deliver them to the autophagic vesicles for clearance from the neurons. Using viral vectors, we will deliver AFN to cells or to mouse brain via specific stereotaxic injections. First, we will test AFN in vitro using 2D and 3D (brain organoid) neuronal cultures derived from iPSCs harboring APP/PSEN1 or α-Syn mutations that drive Aβ or α-Syn aggregation, respectively, and assess the ability of AFN to suppress aggregate formation and neuronal toxicity. Second, we will use mouse models of AD (with a genetic knock-in of human APP mutations that cause Aβ aggregation) and PD (expressing aggregation-prone mutant human α-Syn) to test the therapeutic potential of AFN and its ability to prevent aggregate formation, neurodegeneration, and cognitive or motor deficits in vivo. If successful, the proposed project will have a major impact on the neurodegenerative diseases field, by developing an effective potential gene therapy for AD and PD, and set the foundation for the next steps of preclinical and clinical testing of this suggested therapy. It may also provide a proof-of-concept for therapy development for other neurodegenerative disorders associated with protein aggregation.

NIH Research Projects · FY 2026 · 2022-03

Summary/Abstract Micro-ribonucleic acids (miRNAs) are crucial for normal lung development, lung health, and have been implicated in lung diseases, including Chronic Obstructive Pulmonary Disease (COPD). In addition, recent studies have identified miRNAs as potential therapeutic targets for COPD. miRNAs play an important role in gene regulation. miRNAs act within a complex regulatory structure that involves other biological molecules, including transcription factors, proteins, messenger RNAs (mRNAs), and other epigenetic mechanisms, all of which may be under the influence of genetic variants. A number of genetic variants have been identified as important for COPD through Genome-Wide Association Studies (GWAS). However, substantial computational and methodological challenges impede both linking genetic variants to altered molecular mechanisms and identifying therapeutic interventions that can effectively target these mechanisms for clinical impact. Understanding the complex structure of gene regulation, including by and of miRNAs, and how it is altered in disease or in response to a genetic variant, is critical for developing effective, precision-medicine based therapeutic strategies in COPD. In this project, we hypothesize that network-based methods, which model the collective response of biological molecules, have the potential to identify novel therapeutic strategies for COPD. Our goal is to leverage the predictions made by regulatory network models in a Connectivity Map analysis to identify potential therapeutic interventions for COPD. To accomplish this goal, we will first develop a network approach to model regulation by and of miRNAs using existing mRNA and miRNA expression data from blood samples in COPDGene. We will also generate miRNA expression data in lung tissue from the Lung Tissue Research Consortium (LTRC). We will use these data to assess changes in miRNAs related to COPD and to quantify associations between miRNAs and genetic variants. Co-expression, regulatory, and genetic-association networks will be developed to support multi-Omics predictions relevant to COPD. Network analysis results will be analyzed in the context of drug response profiles from the Connectivity Map to identify potential treatment strategies. The outcome of this project will be a deeper understanding of the regulatory role of miRNAs in COPD, how disease-specific miRNA regulation is altered in the context of COPD GWAS variants, and predictions for novel therapeutic strategies for COPD. The methods developed in the project will also support the broader goal of developing precision-medicine strategies for COPD.

NIH Research Projects · FY 2025 · 2022-03

Project Summary IgE-mediated allergic disease is a growing problem. The pathogenesis of allergic disease requires that immunoglobulin (Ig) E (IgE) molecules be produced against what are otherwise usually innocuous substances. Upon activation in the setting of cytokines such as IL-4 or IL-13, B cells can undergo IgH CSR to IgE. IgE secreted from B lineage cells can, in the presence of cognate antigen, activate mast cells and basophils to release potent inflammatory mediators. While IgE responses can lead to protective immunity as a part of a specialized responses to multicellular pathogens or other noxious threats, they also underlie allergic disease. Allergic disease can be manifest by localized inflammation, or by multiorgan involvement, including deadly systemic anaphylactic reactions via IgE-sensitized mast cell degranulation. Thus, the production and dissemination of IgE play a significant role in dictating the strength and extent of tissue mast cell sensitization. It is therefore critical to understand not only how B cell IgE production and maturation is controlled, but also the principles underlying distribution of IgE from point of origin to distal sites throughout the body. The overall objective of this application is to understand biological aspects of IgE production and dissemination and to gain insights into how this is influenced in allergic disease. Emerging literature and preliminary data from the applicant suggest a general hypothesis that biological constraints cooperate to restrict IgE dissemination under homeostatic conditions, and that accumulation of bone marrow IgE long-lived plasma cells is an aberrancy underlying systemic manifestations of allergic disease. This hypothesis will be tested by pursuing three specific aims, which are: 1) to determine the mechanisms of IgE expression dynamics on IgE B cells; 2) to elucidate mechanisms underlying IgE distribution from point of origin to effector sites; and 3) to characterize IgE plasma cells in allergic patients. Under the first aim, IgE mRNA splicing and IgE surface density will be genetically perturbed to examine the hypothesis that splice bias-mediated dilute IgE BCR density limits independent IgE GC B cell evolution potential. Under the second aim, models of anatomic location-specific allergic challenge will be deployed to examine the degree to which IgE distribution is locally biased, and the role of naïve bystander B cells in this process. Under the third aim, bone marrow aspirations from healthy and allergic individuals will be obtained for IgH isotype-resolved deep sequencing as well as single cell transcriptomics to elucidate the cellular sources and biological properties of IgE in patients with long-standing severe allergies. This contribution is significant because it is expected to elucidate a more complete picture of how IgE responses are regulated. Ultimately, such knowledge has the potential to inform the development of new strategies that will help to reduce the growing problem of allergic disease.

NIH Research Projects · FY 2026 · 2022-02

Abstract Heart transplantation is the optimal therapy for patients with irreversible, end-stage heart disease. However, a several challenges remain to improve allograft and recipient survival. Immunosuppressive agents used to prevent rejection have improved, but they still cannot consistently eliminate acute and chronic rejection, and they are implicated in the pathogenesis of organ failure. New insights into how innate and adaptive immunity contribute to rejection, identification of new therapeutic targets, and novel approaches to promote immune tolerance are major unmet needs in transplantation. Early innate inflammatory responses in the organs (e.g., due to ischemia-reperfusion injuries) enhance acute and chronic heart allograft rejection. Integrins are heterodimeric cell surface receptors involved in immune cell trafficking and signaling; therefore, they are attractive targets to inhibit inflammation, including transplant rejection. The main goal of this project is to elucidate the novel role of β3 integrin in regulating alloimmune responses via control of platelet- and T cell- mediated immunity. Our ultimate objective is to develop new anti-β3 integrin-based strategies to promote engraftment. Our data indicate that β3 integrin-/- mice (β3-/-) show significantly prolonged heart allograft survival in comparison to wild-type (WT) mice, a finding that is associated with reduced CD8+ T cell infiltration into the grafts. We also show that β3 is expressed by activated CD8+ T cells, and that the trafficking of T cells from β3-/- mice is impaired. Notably, targeting β3 integrin also substantially reduces lesions typical of chronic rejection. The β3 subunit is shared by the two integrin molecules, αVβ3 and αIIbβ3, which are expressed by T cells and platelets, respectively. Based on extensive preliminary data, our specific hypothesis is that β3 on both cell types contributes to rejection. In this proposal, we aim to define the relative roles of β3 integrins expressed on platelets (in early promotion of inflammatory responses) and T cells (in enhancement of alloimmunity) in mediating allograft rejection. Furthermore, our targeted delivery method of therapeutics usingnanoparticles (NPs) has emerged as a promising method that increases efficacy and reduces side effects. Here, we have developed first- in-class NPs for targeted delivery of cyclic RGD tripeptides (cRGD) to suppress β3 integrin- mediated recruitment of platelets and T cells for early reduction of chronic rejection, using a murine model of heart transplantation. In this proposal, we present three main aims to determine the roles of αIIbβ3 on platelets (Aim 1) and T cell- expressed β3 (Aim 2) in regulating alloimmunity. In Aim 3, we will perfuse organs prior to transplantation with NPs carrying cRGD to promote graft acceptance.

NIH Research Projects · FY 2026 · 2022-02

Project Summary/Abstract The inflammatory cytokine, interleukin (IL)-17, has emerged as a major player in inflammation-associated, spontaneous, and metastatic colorectal cancer (CRC) models, as well as human CRC. In fact, elevated IL-17 has been negatively correlated with CRC patient survival, and even linked to resistance to both chemotherapy and targeted therapeutics. However, master regulatory mechanisms that control IL-17-IL-17R signaling in CRC remain largely unknown. Key Findings: We recently uncovered a critical role for the microRNA, miR-146a, in preventing colonic inflammation and associated tumorigenesis. Mice deficient in miR-146a (-/-) are highly susceptible to both colitis-associated and spontaneous CRC, which appears to be mediated by enhanced tumorigenic IL-17-IL17R signaling. Mechanistically, our data suggest miR-146a limits intestinal inflammation and CRC by two interlinked mechanisms: 1) miR-146a within myeloid cells inhibits IL-17-inducing cytokines, which restricts IL-17 production; and 2) miR-146a within intestinal epithelial cells (IECs) inhibits tumorigenic IL-17R signaling, which restricts IL-17 responsiveness. Within myeloid cells, miR-146a binds RIPK2, an NOD2 signaling intermediate, to limit myeloid cell-derived IL-17-inducing cytokines, such as IL-23, and restrict colonic IL-17 levels. Accordingly, myeloid cell-specific deletion of miR-146a leads to CRC susceptibility. In addition to inhibiting IL-17, miR-146a directly limits tumorigenic IL-17R signaling within IECs by binding TRAF6. Correspondingly, IEC-specific deletion of miR-146a also confers CRC susceptibility. Importantly, preclinical administration of miR- 146a mimic can ameliorate CRC. Finally, we show that miR-146a appears to analogously target RIPK2 and TRAF6 in humans, including in CRC patients, suggesting miR-146a may also limit IL-17-IL-17R signaling in humans to control CRC. Hypothesis/Goal: We will test our hypothesis that miR-146a protects against CRC by regulating colonic inflammation and tumorigenesis in mice and humans. In Aim 1, we will leverage IEC-specific miR-146a-/- mice, IEC progenitor-specific miR-146a-/- mice, ApcMin/+miR-146a-/- mice, ApcMin/+miR-146a-/-KrasLSL- G12D mice, miR-146a-silenced/overexpressing human IECs, and CRC patient samples to test if miR-146a within IECs targets TRAF6 and inhibits IL-17R-mediated tumorigenesis in mice and humans. In Aim 2, we will leverage myeloid cell-specific miR-146a-/- mice, ApcMin/+miR-146a-/- mice and ApcMin/+miR-146a-/-KrasLSL-G12D mice, miR- 146a-silenced/overexpressing human primary myeloid cells, and CRC patient samples to test if miR-146a within myeloid cells targets RIPK2, where it inhibits NOD2 signaling and IL-17-inducing cytokines, ultimately restricting IL-17 production and CRC in mice and humans. In Aim 3, we will leverage inflammation-associated CRC, spontaneous CRC, and CRC metastasis models to test if miR-146a mimic therapeutically inhibits tumor/chemoresistance-promoting IL-17 pathways, either alone or in combination with chemotherapy. In summary, miR-146a is of unique significance because it constitutes a single target that modulates multiple pathways converging on tumorigenic IL-17 signaling and may offer novel therapeutic points of intervention.