Brigham And Women'S Hospital

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY A diverse repertoire of antibodies is important to protect from a vast variety of infectious threats. Antibodies that can recognize multiple antigenic targets, termed polyreactive antibodies, can provide enhanced B cell receptor (BCR)/antibody recognition capacity. As much as polyreactivity can enhance recognition to thwart invading pathogens, polyreactive antibodies are often autoreactive. The primary BCR repertoire in developing bone marrow B cells is highly polyreactive. As B cells mature, tolerance checkpoints filter the developing BCR repertoire such that polyreactive specificities are rarer in the mature BCR repertoire. Despite its relevance for health and disease, there is a fundamental gap in knowledge regarding how conventional naïve follicular BCR polyreactivity is regulated. The objective of this application is to understand contributory factors influencing the composition of the B cell repertoire. The hypothesis is that the baseline Ig repertoire of conventional follicular B cells is plastic, susceptible to modification of antigen recognition capacity by microbe-regulated tolerance filter passage of polyreactive B cells into the mature naïve follicular B cell repertoire. How microbial symbionts regulate this filter is not known. Supported by preliminary data in the application, one aspect of this modifiable tolerance filter is proposed to take the form of neutrophil-supplied BAFF availability to otherwise counterselected auto/polyreactive B splenic follicular B cells. This hypothesis will be explored with two specific Aims: 1) To elucidate the influence of microbial symbionts on polyreactivity and antigen recognition; and 2) To define the contribution of the IL-17-neutrophil-BAFF axis in influencing microbial symbiont-mediated polyreactivity. The hypothesis of Aim 1 is that baseline follicular BCR repertoire modification unlikely to be due to clonal expansion or somatic mutation. Also posited is that microbe-influenced polyreactivity changes may influence epitope recognition hierarchies upon immune challenge. The hypothesis of Aim 2 is that a contributory mechanism by which microbial symbionts influence baseline Ig repertoires is by increasing neutrophil-supplied BAFF in the spleen, which rescues polyreactive B cells that would otherwise be deleted or anergized at the transitional B cell stage. A high throughput single cell culture system will be used test polyreactivity of B cell subsets from genetically and pharmacologically perturbed mice to examine this hypothesis. This research is significant as it will (i) illuminate how microbial symbionts influence dynamic plasticity of baseline polyreactivity in naïve follicular B cells and downstream antigen recognition, and (ii) identify a new mechanism underlying how symbiotic microbes influence follicular B cell repertoire polyreactivity.

NIH Research Projects · FY 2026 · 2026-06

PROJECT SUMMARY/ABSTRACT Chronic pain presents as a significant source of disability for large numbers of people in the U.S. Of these patients, many suffer from chronic craniofacial and dental pain which is caused by the continuous activation of trigeminal pain sensing neurons (nociceptors). Conditions like trigeminal neuralgia, for instance, are associated with significant morbidity and higher suicide rates. In addition, the current treatments for such persistent pain are limited and often come with debilitating side effects due to their off-target effects. This proposal aims to address this issue by developing an innovative gene therapy strategy designed to selectively silence trigeminal nociceptors while sparing other cell types in the peripheral and central nervous systems. Viral gene therapy is particularly promising for treating intractable craniofacial and dental pain, as it can be administered locally in a single dose to the affected nerve or ganglia, avoiding systemic distribution. Additionally, viral gene therapies are well-positioned for clinical translation, with several already approved by the FDA and many others in trials for neurological disorders. However, current viral vectors lack the ability to distinguish between sensory neuron subtypes, limiting their effectiveness for treating pain. To overcome this, we propose using recent breakthroughs in single-cell transcriptomics and epigenomics to identify cis regulatory elements, such as gene enhancers, that can drive viral gene expression specifically in trigeminal nociceptors. This proposal outlines two specific aims: 1) Identifying gene enhancers uniquely active in trigeminal ganglion nociceptors, and 2) Functionally screening AAVs (adeno-associated viruses) designed for trigeminal nociceptor specificity. In the first aim, we will perform single-cell epigenomic profiling of trigeminal ganglia from mice and humans to prioritize putative enhancers that regulate nociceptor-specific gene expression across species. In the second aim, we will create a library of viruses where each candidate enhancer drives expression of a unique molecular barcode, followed by in vivo screening using single-nucleus RNA sequencing to identify barcodes expressed primarily in trigeminal nociceptors. Successful completion of this project will produce nociceptor-specific viral tools for pain research. We also anticipate that these enhancers, which would be conserved across species, will enable similar cell-type specificity in humans. Ultimately, these vectors can be engineered to express inhibitory ion channels (e.g., chemogenetic or optogenetic), offering researchers and patients a non-opioid, on-demand solution for craniofacial and dental pain relief.

NIH Research Projects · FY 2026 · 2026-06

Migraine is a unique and severe form of headache. It is also common, experienced by up to 30% of women with age-related prevalence 2-3x higher than men, and is a leading cause of disability worldwide. Migraine has a broadly neurovascular pathophysiology that may be treated by recently introduced drugs targeting the calcitonin gene-related peptide (CGRP), the CGRP receptor, or the serotonin receptor (HTRA11F). However, these drugs remain ineffective or contraindicated for many migraine sufferers, highlighting the need for additional therapeutics. Disability due to migraine is compounded by an approximate 1.5-fold elevated risk of ischemic stroke that is further elevated among individuals who experience migraine with aura, a form of migraine typically experienced as a reversible visual disturbance preceding the headache. The cause of such strokes is unknown but may involve intrinsic properties of the vasculature and/or microembolism. Both the identification of critically needed new therapeutic targets and a better understanding of the elevated ischemic stroke risk would be advanced by addressing gaps in knowledge about the pathophysiology of migraine susceptibility. The International Headache Genetics Consortium (IHGC) recently (in 2022) reported a new genome-wide association study (GWAS) that identifies 123 susceptibility loci for migraine with a strong aggregate signal related to vascular biology and also evidence of neurological functions. Here, we propose computational and experimental approaches to investigate specific causal mechanisms of migraine related to the genetic signals in this GWAS as well as in a proposed new genetic analysis of ischemic stroke among individuals with migraine. In Specific Aim 1, we will perform the new GWAS of ischemic stroke among individuals with migraine using existing large cohorts and biobanks to identify key risk loci and therefore genes underlying these events. In Specific Aim 2, we will investigate the impact of loci in the IHGC 2022 migraine and Specific Aim 1 GWAS’s on patterns of coordinately regulated, cell-specific transcription (“programs”) using two approaches: 1) Newly available computational methods that leverage public single cell expression data, and 2) a recently proven experimental strategy that perturbs the RNA expression at the GWAS loci in vascular endothelial and smooth muscle cells. Specific Aim 3, we would use computational and experimental approaches first to investigate overlap of targets of recent CGRP and ditan therapies with inferred transcriptional programs or biological pathways of migraine and strokes to better understand the mechanisms of these new drugs. Then, we will use similar methods to investigate whether existing drugs may be candidates for reducing the burden of migraine or stroke through repurposing. Thus, we propose three independent but complementary Specific Aims to advance mechanistic understanding of the pathophysiology of migraine and the associated risk of ischemic stroke, toward elucidating the action of existing treatments and potentially identifying novel therapeutic approaches.

NIH Research Projects · FY 2026 · 2026-06

Project Summary/Abstract Systemic sclerosis (SSc) is a chronic autoimmune disease characterized by microvascular damage, immune dysregulation and multiorgan fibrosis. Lung disease manifesting as interstitial lung disease (ILD, which can occur within 2-5 years of SSc diagnosis); and pulmonary arterial hypertension, PAH, account for 60% of SSc-related deaths. Although no true disease modifying agent is available, immunomodulation is the backbone of therapy, while antifibrotics for SSc-ILD can be used. Given the complex and variable nature, investigating identification of patients at risk of SSc-ILD to reduce morbidity and mortality is warranted. Dr. Tukpah’s research group designed a large, well characterized electronic health record (EHR) based cohort of patients with SSc to use longitudinal data and linked biorepository specimens for analyses. Dr. Tukpah’s initial analyses demonstrated age at SSc-ILD, timing to SSc-ILD diagnosis and emergency room rates differed among patient groups. Some patient populations had the youngest age at SSc-ILD diagnosis and impaired lung function at baseline, which may be associated with certain autoantibody profiles. Fully characterized risk factors for SSc- ILD remain unknown, phenotyping lung function decline requires further investigation and tools to predict which patients will develop SSc-ILD are lacking. Furthermore, circulating diagnostic biomarkers are not available. Collectively, this proposal aims to define previously uncharacterized signatures of high risk of SSc-ILD; predict incident SSc-ILD, especially early onset; and identify plasma protein biomarkers associated with SSc-ILD. In the first aim, Dr. Tukpah will build models (with biological, environmental, clinical data) to analyze risk factors for SSc-ILD and lung function decline using physiological parameters of disease progression. She will also create supervised and unsupervised machine learning models to predict incident SSc-ILD. In the second aim, she will conduct biobank plasma proteomic analyses to detect differential protein levels in patients with SSc- ILD which can ultimately guide diagnostic biomarker evaluation and promote earlier detection. Lastly, in an external cohort, she will validate models from Aim 1 and plasma protein findings from Aim 2. Dr. Tukpah will perform this work in the Division of Pulmonary and Critical Care Medicine, at Brigham and Women’s Hospital (BWH), a core teaching hospital of Harvard Medical School. Dr. Tukpah’s mentors are Dr. David Bates, an expert in clinical informatics, and Dr. Gary Matt Hunninghake, an expert in the field of early ILD characterization. With the guidance of her mentors and scientific advisory committee, Dr. Tukpah has developed a comprehensive five-year training program to advance the skills needed to complete the project and become an independent investigator with expertise in pulmonary biomedical informatics. Dr. Tukpah is committed to an academic medicine career. As a physician-scientist she will use knowledge and skills gained during this project to better understand risk phenotypes and predict and ultimately diagnose incident disease.

NIH Research Projects · FY 2026 · 2026-06

Project Summary Neutrophils play a critical role in innate immunity and host defense against invading microorganisms, but also contribute pathogenically to a number of non-infectious conditions including asthma, multiple sclerosis, rheumatoid arthritis, gastritis, inflammatory bowel disease, cancer, and ischemia-induced tissue damage. Disentangling the beneficial and harmful activities of neutrophils has proven exceptionally difficult, as they present with varied and often opposing physiological functions that are influenced by the timing and location of the inflammatory response. Recent studies have revealed that neutrophils are not a homogeneous cell population; instead, they display a diverse array of functional states, phenotypes, and lifespans. This offers an opportunity to develop tailored therapeutic strategies by targeting distinct neutrophil subsets or inducing specific functional states in neutrophils. The theme of this project is to uncover the nature and function of neutrophil heterogeneity, a fundamental question in neutrophil biology. The hypothesis is that the neutrophil compartment is a collection of cell phenotypes, states, and sub-lineages capable of distinct functions. Understanding how these subpopulations arise and function will define new targets for neutrophil manipulation, establishing specific lineages as vehicles for clinical intervention. Dr. Luo (Project 1) will investigate the origin and characterize the functional state of lung interstitial neutrophils (LINs). Dr. Mayadas (Project 2) will investigate the origin, regulation and trafficking of a subset of spleen-derived neutrophils that acquire antigen- presenting cell (nAPCs) properties following engagement of their FcγRs. Dr. Nigrovic (Project 3) will test the hypothesis that emperipolesis of neutrophils by megakaryocytes in the bone marrow modulates the phenotype of neutrophils and thereby drives neutrophil heterogeneity. Finally, Dr. Hidalgo (Project 4) will evaluate the hypothesis that neutrophils with antimicrobial and inflammatory properties have different origins and will further characterize the properties and origin of neutrophils that mediate inflammation. The four research projects will be bolstered by a unique centralized Bioinformatics Core led by Andres Hidalgo, which is particularly valuable in this collaborative research project as it will enhance data quality, accessibility to expertise, efficiency, collaboration potential, and overall scientific rigor. An Administrative Core led by Hongbo Luo will coordinate and direct the collaborative, multi-disciplinary, interactive, and synergistic research activities of these projects and cores to create a cohesive unit. Collectively, the four projects proposed here, supported by the two cores, will work together to form a unified and productive program to comprehensively elucidate the origin and cellular mechanisms driving the heterogeneity of neutrophils and their reprogramming in health and disease. This work will lay the foundation for neutrophil subset-specific therapeutic strategies in the treatment of infectious and inflammatory diseases.

NIH Research Projects · FY 2026 · 2026-06

ABSTRACT Premature rupture of membranes (PROM) and preterm PROM (pPROM) affect up to 10% and 4% of pregnancies globally, respectively, leading to significant maternal and fetal health risks if not detected and managed promptly. These risks include maternal chorioamnionitis, neonatal sepsis, umbilical cord prolapse, placental abruption, and preterm labor. Current detection methods, such as the Nitrazine test and immunoassays for biomarkers like PAMG-1 and IGFBP-1, are effective but are often costly, invasive, and depend on the mother's ability to identify abnormal vaginal discharge. Such dependency on patient recognition is problematic, especially in persistent low-volume leakage where symptoms are easily mistaken for normal discharge, delaying diagnosis and treatment. These challenges are further compounded in rural or underserved areas, where limited access to healthcare can delay diagnosis and increase risks. Our research aims to develop a novel colorimetric detection system that functions as a vaginal insert for the early identification of premature rupture of membranes (PROM) and preterm PROM (pPROM). This insert is specifically designed to distinguish amniotic fluid from other vaginal discharges, offering an at-home, minimally invasive solution that integrates into a patient’s daily routine. By utilizing a pH-sensitive polymer, the device reacts to the presence of amniotic fluid by releasing a biocompatible dye, causing a visible color change in vaginal discharge. This allows for early detection, even with small leaks, enabling patients to seek timely medical care without frequent clinical visits. The system leverages the design of pessary devices, which are already widely used by pregnant women for long-term use, ensuring comfort and improving patient compliance. Placing the device close to the uterus maximizes contact with amniotic fluid while minimizing interference from other secretions. The device uses FDA-approved materials to ensure biocompatibility and safety for both mother and fetus. In contrast to costly and time-consuming biomarker assays, this solution provides a simple, cost-effective alternative by utilizing the pH difference between amniotic fluid and vaginal discharge. Ultimately, this system offers a user-friendly, accessible, and highly sensitive method to detect PROM and pPROM, improving maternal and fetal health outcomes, especially in low-resource settings. Aim 1: Optimize the formulation of a pH-responsive polymer with additives to ensure stability in an acidic vaginal environment while maintaining high sensitivity to amniotic fluid. Aim 2: Refine device designs, including geometric configurations and material placement, to enhance detection sensitivity and specificity, minimizing false positives and negatives. Aim 3: Assess the biocompatibility, stability, and effectiveness (i.e., sensitivity and selectivity) of the prototype devices for detecting amniotic fluid leakage using a mouse model

NIH Research Projects · FY 2026 · 2026-06

Project Summary Epstein-Barr virus (EBV) causes ~350,000 cases of various cancers including several B cell malignancies, nasopharyngeal carcinoma and 10% of the gastric cancer. EBV also causes infectious mononucleosis and is linked to autoimmune diseases including multiple sclerosis. EBV infects both B cells and oral epithelial cells. The B cell entry mechanism is well characterized. The epithelial cell entry mechanisms just started to be unraveled. To identify additional epithelial entry receptors, we used genome wide CRIPSR screen and identified additional EBV entry receptors DSC2/3. However, little is known about how do these new entry receptors allow EBV entry. Our specific aims are 1. Determine the mechanisms through which cell-free EBV exploits DSC2/3 to enter epithelial cells. 2. Determine the mechanisms through which EBV exploits DSC2/3 in cell-cell contact infection to infect epithelial cells. 3. Determine the structural and functional interaction between EBV glycoproteins and DSC2/3. Since EBV entering host cells is a critical step for EBV to cause diseases, understanding the mechanisms through which EBV enter cells will help us to understand the disease pathogenesis. This also may identify potential targets for perturbation and treatment, allowing rational design to block EBV entry into host cells to prevent EBV associated diseases

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY/ABSTRACT In chronic obstructive pulmonary disease (COPD), females are typically diagnosed at younger ages with more severe disease, less smoking exposure, and more comorbidities. Yet the role of the X chromosome in COPD remains largely unexplored, particularly regarding incomplete X chromosome inactivation (XCI), which can lead to gene dosage imbalances and sex-specific effects of genetic variation in regions that escape XCI. Recently, our group completed an X chromosome-wide association study (XWAS) in COPD and identified X-linked variants near escape genes associated with COPD risk and related traits. Using network analyses, we have further demonstrated sex-specific patterns of gene regulation, especially in the transcriptional targeting of XCI escape genes in lung and blood. We hypothesize that COPD heterogeneity is partially driven by genetic and epigenetic variation on the X chromosome and incomplete XCI. To test this, we will integrate X chromosome multi-omic data—including whole genome sequencing, DNA methylation, and gene expression—from lung and blood samples across four cohorts (COPDGene, LTRC, LTCOPD, and ECLIPSE). We will use methylation Quantitative Trait Loci (mQTL) and colocalization analyses to identify mechanisms by which X chromosome genetic variants influence COPD susceptibility and heterogeneity through epigenetic regulation (Aim 1). We will associate X chromosome genetic variants with gene expression through an expression Quantitative Trait Loci (eQTL) analysis (Aim 2). To comprehensively map these regulatory mechanisms, we will use innovative gene regulatory network approaches that account for X chromosome dosage (Aim 3). This integrative framework will enhance our understanding of X chromosome gene regulation in COPD, and identify molecular mechanisms underlying disease heterogeneity. We will examine XCI escape genes in the context of mQTL, eQTL, and regulatory networks, and test for changes between COPD cases and controls. Identifying variations in XCI patterns in disease will give insights into the mechanisms associated with both disease development and sex differences in COPD. Ultimately, this project will provide a comprehensive map of XCI escape and X-linked gene regulation in COPD, X chromosome genetic and epigenetic variation that contribute to COPD susceptibility and heterogeneity, and establish new framework for including allosomes in network analyses. This new K01 award in response to NOT-OD-25-115 will enable Dr. Lopes-Ramos to advance her skills in epigenetics, statistical genetics, pulmonology, integrative omic and network analysis, and develop as an independent researcher in integrative respiratory omics and network medicine, with continued mentored training and a clear trajectory toward research independence.

NIH Research Projects · FY 2026 · 2026-05

PROJECT ABSTRACT Type 2 diabetes affects 16.5 million U.S. adults aged ≥65, nearly one-third of the senior population, and is associated with higher rates of hypoglycemia, cardiovascular events, and excess healthcare costs. Continuous glucose monitors (CGMs) offer the potential for improving glycemic control through real-time data, yet CGM adoption and effectiveness remain poorly characterized in older adults. Evidence from trials is limited or inconsistent in this population, and few studies evaluate how clinicians interpret and act on CGM data in routine care, representing a critical gap in addressing therapeutic inertia. This proposal applies a structure-process-outcome framework to evaluate age-related differences in CGM implementation, clinical outcomes, and care processes across two large healthcare systems (Mass General Brigham and the University of California). In Aim 1, we will assess how CGM-derived metrics (e.g., time in range, time below range) influence clinician behavior using mixed-effect models and group-based trajectory modeling to identify glycemic phenotypes linked to treatment decisions. In Aim 2, we will use difference-in-differences and mixed-effect modeling to compare outcomes (HbA1c, blood pressure, LDL, weight, hypoglycemia) and care processes (e.g., medication changes, referrals) between CGM users and non-users across age groups. In Aim 3, we will apply the Consolidated Framework for Implementation Research and rapid ethnography to identify structural and multilevel factors influencing CGM adoption among older adults. This work is innovative in its integration of CGM and EHR data to assess real-world therapeutic inertia, application of trajectory modeling to reveal age-related glycemic patterns, and use of implementation science to identify system-level levers for change. Findings will provide actionable evidence to guide age-responsive CGM implementation strategies, reduce disparities, and improve outcomes for older adults with Type 2 diabetes.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY (24/30 lines) This proposal seeks to improve the management and survival of patients with localized renal cell carcinoma (RCC) at high risk of relapse by optimizing the utilization of anti-PD-1 therapy in the neoadjuvant/adjuvant settings. Despite surgical management, approximately 30% of patients with localized RCC develop metastatic disease, which remains largely incurable. Anti-PD-1-based therapies have shown efficacy in treating metastatic RCC, but inconsistent clinical trial results indicate a need for more precise patient selection for the implementation of these therapies in the neoadjuvant and adjuvant settings. Our multidisciplinary team has substantial experience in translational kidney cancer research and proposes to utilize the unique of collection of tissue and blood/plasma specimens from the EA8143 PROSPER (NCT03055013) trial to achieve the following aims: Aim 1. To identify tissue-based biomarkers that can predict benefit from neoadj/adj anti-PD-1 therapy in localized RCC. We plan to analyze baseline (untreated) tissue samples to develop biomarkers, including somatic genetic alterations and immune markers, that predict benefit from neoadjuvant/adjuvant nivolumab therapy relative to observation. Aim 2. To identify circulating plasma biomarkers that can predict benefit from neoadj/adj anti-PD-1 therapy in localized RCC. We plan to analyze baseline (untreated) plasma samples to develop non-invasive biomarkers, including circulating levels of kidney injury molecule-1 (KIM-1) and chemokines/cytokines, that predict benefit from neoadjuvant/adjuvant nivolumab therapy relative to observation. Aim 3. To evaluate the immunomodulatory effects of neoadjuvant anti-PD-1 therapy in localized RCC. We plan to test the hypothesis that neoadjuvant nivolumab leads to a reinvigorated anti-tumor immune response that correlates with improved outcomes following neoadjuvant/adjuvant nivolumab therapy. We plan to further conduct exploratory analyses aimed at (i) identifying determinants of resistance to nivolumab therapy, and (ii) characterizing putative tumor-specific T cells clones in relation to nivolumab therapy. Overall, this project is likely to generate clinically meaningful results that have the potential to improve the outcomes of patients with localized RCC who are at high risk of recurrence and death.

NIH Research Projects · FY 2026 · 2026-05

Abstract Each muscle of our body is endowed with a specific identity defined by its location, shape, organization and function. This identity is largely dependent on the HOX family of genes which controls the regional anat- omy of tissues along the body axis 3,4. Despite nearly half a century of research, our understanding of HOX genes’ function in development remains limited, primarily due to the significant functional redundancy within this gene family. The challenge lies in the inability, to date, to generate a vertebrate embryo completely devoid of HOX genes due to the complexity of the genetics involved and allele lethality. So far only very limited loss of HOX functions have been achieved in vertebrates, revealing only part of their developmental roles. We have developed new human iPS-based in vitro models recapitulating early stages of skeletal muscle development with remarkable accuracy 2,5. These cell culture systems exhibit the expected collinear expression of HOX genes, presenting an excellent platform for studying the functions of this gene family in controlling skeletal muscle development. A significant advantage of these in vitro models is their ability to circumvent the lethality associated with HOX clusters deletions in model organisms. This application proposes leveraging our human iPS-based in vitro models to investigate the cellular and molecular mechanisms underlying HOX gene function during development, focusing on somitic derivatives such as skeletal muscle as well as motoneurons. Using CRISPR-CAS9, we have created a series of HOX clusters deletions in a human iPS line, including one (HOXKO) harboring a complete deletion of all HOX clusters. This line can differentiate to the paraxial meso- derm fate but fails to form epithelial somites and cannot generate skeletal muscle under a standard myogenic differentiation protocol 6. These defects are not observed when three out of four clusters are deleted and thus require the full removal of HOX genes. Notably, these defects can be rescued by inducible expression of a sin- gle HOX gene from different paralog groups. We also showed that retinoic acid (RA) signaling, which has re- cently been implicated in somite epithelialization7 is downregulated in HOXKO somitoids. Somite formation can be rescued by exogenous RA treatment in HOXKO cells. These results suggest central roles of HOX genes in somitogenesis and myogenesis, previously masked by functional redundancy, which we propose to study here. Our proposed experiments combine sophisticated live imaging to transcriptome and epigenome analysis to identify the cellular and molecular basis of HOX function during skeletal muscle development. Additionally, we seek to investigate whether HOX genes act as pioneer factors and identify the targets they regulate during my- ogenesis. These experiments provide a unique opportunity to unravel the regulatory logic underlying HOX function in the intricate control of patterning and morphogenesis of skeletal muscle.

NIH Research Projects · FY 2026 · 2026-05

Project Summary Epstein-Barr Virus (EBV) causes infectious mononucleosis, lymphomas and lymphoproliferative diseases in HIV infected and immune suppressed people and is linked to autoimmune diseases. EBV converts Resting B Lymphocytes (RBLs) to continuously proliferating Lymphoblasts Cell Lines (LCLs) by expressing EBV nuclear antigens (EBNA) and latent membrane protein 1 (LMP1) that activates NF-kB. Since LCLs express the same EBV proteins as some EBV cancers, EBV conversion of RBLs to LCLs is therefore a relevant model that can be genetically manipulated to investigate EBV's role in growth transformation. LCL growth depends on EBNA2, EBNALP, EBNA3A, EBNA3C and LMP1. Recently, we found that all the essential EBNAs and LMP1 activated NF-kB subunits converge to EBV super-enhancers (ESE) that have extraordinary H3K27ac signals. ESEs govern the expression of key oncogenes that drive LCL growth and are more sensitive to perturbations than average enhancers. To further characterize the molecular composition and their functional roles in ESEs, we will (1) Identify DNA elements essential for ESE activity, (2) Determine the mechanisms through which ESE proteomic components affect ESE activity, and (3) Determine the functional roles of ESE eRNAs. We will use Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) based assays to identify the DNA elements essential for ESE to loop to their direct target genes and proteins essential for ESE activity. We will test the effect of eRNA knock down on host transcription and looping factor DNA binding. The experiments here in use integrative approaches to elucidate the molecular mechanism by which ESEs activate key oncogenic drivers. These studies will identify opportunities for therapeutic intervention.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY: Background. Traditional drug development is slow, costly, and inefficient, with over 90% of drugs failing in clinical trials. Personalized medicine also faces challenges in accurately matching patients with optimal treatments. "Phase-0" microdosing, involving the delivery of tiny drug doses directly into target tissues, has the potential to transform both drug development and personalized drug selection by directly evaluating drug effects in patients without systemic toxicity and rapidly identifying the best treatment for each individual. Our team is leading the clinical translation of an implantable microdevice (IMD) that provides Phase-0 microdose readouts of multiple drugs in a single patient. IMDs are placed into diseased tissue (most commonly tumors) and subsequently release drugs into spatially discrete microscopic regions. The devices and surrounding tissue are then removed surgically and drug effects are characterized and compared using a multi-omic spatial analysis pipeline. Preclinical and first-in-human IMD trials have been highly promising with unprecedented drug insights. However, surgical resection required for IMD analysis is not feasible or safe in most deep anatomic locations. This has prevented IMD validation and use in most settings where they are needed, including neoadjuvant and aggressive metastatic cancer. A less invasive and safer approach is needed to enable broad clinical application. More specifically, there is a critical and currently unmet need for a nonsurgical interventional device to deliver drug microdoses and subsequently biopsy drug-containing tissues for Phase-0 measurements. Proposal and impact. In Aim 1, we will develop an interventional drug response assay (IDRA) device that will broadly enable biopsy-based Phase-0 measurements from any percutaneously accessible site similar to routine percutaneous biopsies, with similar outpatient workflow and low risk. In Aim 2, we will validate IDRA safety and feasibility in prostate and ovarian cancer pre-clinical models (Aim 2). This will enable clinical investigational trials in neoadjuvant and advanced metastatic prostate and ovarian cancer, two settings where oncologists have expressed a critical and urgent need. In a parallel Aim 3, we will integrate label-free optical imaging into the IDRA and evaluate feasibility to image real-time drug effects directly in tumors (Aim 3). Aim 3 will lay the groundwork for our long-term goal of dynamic in-vivo optical Phase-0 imaging, which would provide unprecedented capability to directly monitor drug response and resistance over time. In the long term, these aims have the potential to broadly transform drug development and personalized treatment selection in diseases throughout the body.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY The electronic MEdical Records and GEnomics (eMERGE IV) Network is a national network organized and funded by the National Human Genome Research Institute (NHGRI) that combines DNA biorepositories with electronic health record (EHR) systems for large scale, high- throughput genetic research in support of implementing genomic medicine. In 2020, eMERGE IV launched a study of genomic risk assessment and return of genomically informed risk assessments (GIRA) with healthcare recommendations for 11 common diseases to a cohort of 23,840 participants (including 4,255 children) from primary care practices across 10 institutions. The primary outcome is analysis of uptake of pre-specified care recommendations (provider action, participant action) across all conditions at 12-months post-GIRA return in high- risk vs. not high-risk participants as defined by the Network-designed statistical analysis plan. Mass General Brigham (MGB) has been an active and engaged eMERGE IV site with specific expertise in EHR data extraction, quality control (QC) processes, and use of large language models (LLMs) to define EHR outcomes. We enrolled and completed GIRA return for 2502 MGB participants, uploaded the results to the EHR, surveyed providers, implemented post- RoR participant surveys, and extracted and QC’d the 6-month EHR outcomes. Extraction of 12- month EHR outcomes is underway and expected to finish in April 2026. Our site has provided Network leadership and was deeply engaged with the other sites and Network leadership throughout the study. In this proposal we request a one-year extension of funding for a “Year 7” from 05/01/2026 to 4/30/2027. This extension will allow the MGB eMERGE IV site to QC and harmonize 12-month outcome data, analyze whether returning high risk GIRA leads to new preventive and screening healthcare actions, and disseminate results through manuscripts to inform implementation of genomic medicine in healthcare for adults and children. We intend to work with the NHGRI leadership and Network sites to continue to discover and publish the learnings from the eMERGE IV genomic risk implementation study following the plans for an extension year that we collaboratively designed as a network.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY/ABSTRACT Cushing’s syndrome results from chronic exposure to excess glucocorticoids, which can be from an endogenous source of autonomous cortisol production or from exogenous administration of glucocorticoids. Chronic hypercortisolism leads to multiple morbidities, including metabolic syndrome and infertility, as well as increased mortality. These comorbidities may not be resolved even after successful treatment and remission. The neuronal mechanisms regulating fertility are tightly linked to energy balance. Hypothalamic neurons expressing agouti-related peptide (AgRP) are activated after fasting and have been proposed as the link between metabolism and reproduction. Increased activity of this hunger pathway leads to reduced GnRH/LH pulse generation and subsequently to impaired fertility. Chronic glucocorticoid exposure on the other hand, which is known to cause hyperphagia and infertility, has been shown to increase AgRP activity, as well. This proposal aims to explore whether glucocorticoids act directly on AgRP neurons to mediate the metabolic and reproductive effects. First, I will elucidate the role of AgRP neurons in these glucocorticoid-induced effects. Using a chemogenetic approach I will determine if the inhibition of AgRP neurons can prevent or ameliorate reproductive and metabolic impairments. Then, I will generate a mouse model with conditional deletion of glucocorticoid receptor (GR) in AgRP neurons to explore their resistance to developing Cushing’s phenotype under chronic glucocorticoid treatment. In Agrp-cre mice treated with glucocorticoids I will selectively delete GR in AgRP neurons (via CRISPR approach) after developing the Cushing’s phenotype to improve fertility and obesity. Finally given the fact that Cushing’s syndrome is more prevalent in women and these complications can persist in many patients even after remission, I hypothesize that chronic glucocorticoid exposure will mediate transcriptomic and epigenetic modifications within the GR-gene regulatory network of the arcuate nucleus of hypothalamus in a sex-dependent manner and among the affected genes will be genes associated with energy balance and reproduction. Using RNA seq analysis and a highly sensitive fluorescent in situ hybridization I will try to identify differentially expressed genes within the AgRP neurons after chronic glucocorticoid exposure. To further investigate potential long-lasting GR conformational changes, I will perform chromatin immunoprecipitation sequencing analysis. In summary, the overarching goals of this proposal are to elucidate the precise mechanisms by which chronic glucocorticoid exposure affects metabolism and reproduction, focusing on the role of AgRP neurons, and to advance our comprehension of the transcriptomic and epigenetic alterations induced by chronic glucocorticoid exposure. Successful completion of this project may identify potential therapeutic targets to improve the quality of life for patients with Cushing’s syndrome. Furthermore, it may discover unique biomarkers in genetically susceptible individuals, thereby aiding in the prediction of patients at risk of developing these comorbidities.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY/ABSTRACT This proposal addresses an important, unmet need in cancer symptom management: cancer patients are routinely impacted by chronic symptoms caused by their disease and also by cancer-directed treatment. These long-term sequalae of cancer and cancer-therapy can be devastating to patients' physical, psychological, social, and financial wellbeing. However, high-quality symptom care is often limited by patient accessibility, patient and clinician education, and communication with and among the care team. Chatbots for symptom monitoring and management have been shown to improve patient empowerment, healthcare utilization, and outcomes, but have previously been limited to rules-based systems which can be inflexible, require significant manual effort to develop, and cannot easily adapt to new guidelines and user preferences. The overarching objective of this proposal will be to advance new methods and benchmarks for the development of guideline-grounded chatbots for improved cancer symptom education and communication. Our central innovation will be the research and development of efficient, extensible, and user-centered methods that leverage advances in large language models (LLMs) to convert clinical resources into usable, interactive technologies with improved performance and accessibility. In Specific Aim 1, we will improve the factuality of LLM question-answering about symptoms, which we will approach by developing new methods to link LLMs with knowledge in multi-modal clinical practice guidelines. In Specific Aim 2, we will investigate high-fidelity methods for LLM-based text simplification of recommendations in guidelines and the electronic health records to accommodate the different needs of end- users. In Specific Aim 3, we will study the needs, perceptions, concerns, and ethical considerations of patients and clinicians, which will inform technical developments is Specific Aims 1/2 and guide user-centered design for chatbot interfaces. The resulting interface will be evaluated in empirical studies. Patient and clinician stakeholders will be centered throughout all aims of our research. This work will be highly significant and innovative because it uses advances in artificial intelligence to amplify the availability of reliable information resources for cancer symptom care. These methods may thereby improve cancer outcomes and quality-of-life, while providing broad generalizable insights for the use of artificial intelligence-based information and education resources across biomedical fields.

NIH Research Projects · FY 2026 · 2026-05

Project Summary/Abstract Mutations in leucine-rich repeat kinase 2 (LRRK2) are linked to familial Parkinson’s disease (PD) with Lewy body and/or tau pathologies and are also associated with idiopathic PD, dementia with Lewy bodies (DLB), and tauopathies. Despite the importance, pathogenic mechanisms of LRRK2 mutations remain unresolved. Because of early discoveries that LRRK2 mutations increase its kinase activity, over the past two decades major pharmaceutical companies and academic labs worldwide have focused on the development of kinase inhibitors. However, LRRK2 has several other functional domains, suggesting that it has additional functions beyond kinase. Furthermore, our genetic studies revealed unexpectedly that LRRK2 and its functional homologue LRRK1 play an overlapping, essential physiological role in the protection of dopaminergic (DA) neurons in the substantia nigra pars compacta (SNpc) and DA terminals in the striatum during aging. Moreover, development and multidisciplinary analysis of DA neuron-specific conditional knockout mice further demonstrated that this critical, relevant physiological role is cell autonomous to DA neurons in the SNpc, highlighting the importance of LRRK2 in protecting DA neurons from degeneration in the aging brain. These findings suggested that pathogenic mechanisms of LRRK2 mutations might be more complex than simply enhancing kinase activity. Importantly, there has been no experimental evidence demonstrating that increased LRRK2 kinase activity causes DA neurodegeneration in the aging brain. In this application, we propose to investigate whether mitochondrial and/or autophagy-lysosomal dysfunction underlies LRRK2-dependent DA neuronal survival in a cell autonomous or non-cell autonomous manner. We will further investigate the impact of two unique, important LRRK2 mutations, R1441C and G2019S, using the physiologically relevant knockin alleles compared to the null alleles, on DA neuronal survival during aging, mitochondrial and autophagy-lysosomal function in DA and cortical neurons, and whether LRRK2 mutations affect cognition, synaptic plasticity, and mitochondrial calcium homeostasis in the cerebral cortex. Completion of the proposed studies will test whether increased kinase activity by LRRK2 mutations causes DA neurodegeneration and will provide new insights into pathogenic mechanisms of LRRK2 mutations in these human diseases as well as LRRK2-based therapeutic development.

NIH Research Projects · FY 2026 · 2026-04

PROJECT SUMMARY The two-day scientific conference entitled, "The Science of Tai Chi & Qigong as Whole Person Health: Deepening our Physiological Understanding of Mind-Body Integration” will be held at the Harvard Medical School in Boston, MA on April 30-May 1, 2026. The key goals of this conference are to provide a forum for active dissemination of the current evidence base of Tai Chi, Qigong and related mind-body practices (TCQMB), and exploration and thoughtful dialogue on cross-relevant topics integral to the understanding of physiological mechanisms and the impact of mind-body movement in whole person health within the future of healthcare. The primary sponsor of this conference will be the Osher Center for Integrative Health at Brigham and Women’s Hospital and Harvard Medical School. This scientific conference will build upon the success and momentum of the first-of-its-kind, inaugural conference in 2023 that showcased TCQMB in a rigorous academic setting. This upcoming 2026 conference aims to attract a heterogeneous group of international research scholars, practitioners, healthcare administrators and policymakers to engage in a deeper dive into the science. The conference program will include four plenary panel sessions from leaders in the field focused on the following themes in the context of TCQMB: 1) Aging Brain, Aging Body, and Mind-Body Health; 2) Leveraging Technology in Research and Implementation; 3) Cancer and Integrative Oncology; and 4) Research at the Frontier: Fascia, Fluids, Mitochondria, and Biofields. These plenary panels will each include complementary 20-30 minute plenary presentations followed by an extended panel discussion including audience participation. A Call for Sessions will solicit additional concomitant one-hour ‘breakout’ symposia sessions with priority to the following topics: i. remote delivery/virtual platforms/telehealth, use of wearable sensors, and other biotechnology; ii. Impact of TCQMB on neural, physiological, and biomechanical biomarkers of interoception and resilience; iii. TCQMB for specific populations (e.g., stroke, pediatrics, cancer); iv. TCQMB as part of multicomponent interventions; v. Citizen science, big data, and AI. In addition, we will solicit high-quality scientific abstracts with opportunity for both junior and established investigators to present oral and poster presentations. An international Scientific Review Committee, composed of external reviewers, will employ a rigorous peer review process to ensure rigor and relevance of research. Marketing efforts will aim to target a broad audience nationally and internationally with conference content relevant across the broad field of mind-body medicine. Our overarching Conference Aims are 1) To provide an international forum for researchers to disseminate scientific findings relevant to TCQ and related mind- body practices, assess the evidence base in the context of whole person health, and shape the future research agenda, and 2) To foster interdisciplinary dialogue and collaboration to develop innovative strategies that address current challenges in mind-body research, dissemination and implementation.

NIH Research Projects · FY 2026 · 2026-04

PROJECT SUMMARY: Mycobacterium tuberculosis (Mtb) is the leading cause of death worldwide due to a single pathogen. Mtb is exceptionally reliant on specialized lipids, yet little is known about how lipids and lipid- linked glycans transit across the membranes and aqueous periplasm of mycobacteria. My objective is to reveal how Mtb controls the localization and effects of important lipidic antigens, virulence factors, and cell envelope components. I found that one of the primary mycobacterial lipid transport systems, Mce4, is not restricted to exogenous lipid import as previously thought, but maintains steady-state levels of hundreds of native mycobacterial lipids. I will define how periplasm-spanning Mce channels mediate directional lipid transport in Mtb, using unbiased lipidomic methods, a new reverse micelle separation technique, and collisional mass spectrometry. This will push the field beyond previous studies of selected exogenous lipids, and will identify the roles that Mce channels play in native lipid transport within laboratory and clinical strains of Mtb. I also found that the putative transport lipid mannosyl phosphomycoketide (MPM) is covalently linked to one or more Mtb glycans, and that the gene pks12—which is required for the synthesis of MPM—is important for glycan localization. This suggests that MPM is at the center of an undiscovered lipoglycan transport mechanism. I will uncover this mechanism through chromatography and mass spectrometry-based identification of MPM-linked glycans. Through the same methods along with electron microscopy and glycan labeling, I will determine the effects of pks12 on exported Mtb glycans. I will finish by determining the role that pks12 and glycans play in a growth defect I discovered, thereby providing a mechanistic basis for the likely in vivo essentiality of pks12 in human disease. This proposed investigation of understudied Mtb lipid and lipoglycan transport will address a critical knowledge gap that hinders development of tuberculosis drugs, vaccines, and diagnostics. I will conduct the K99 phase of this award as a postdoctoral research fellow in Dr. D. Branch Moody’s group at Brigham and Women’s Hospital. The Moody lab is exceptionally well equipped for mass spectrometry analysis of Mtb lipids and cell envelope components. I will rely on advisors including Dr. Laura Kiessling to support my studies of glycan and lipoglycan transport, and will leverage the scientific networks of Harvard Medical School, its teaching hospitals, the Harvard T.H. Chan School of Public Health, and the broader Boston research community. In my mentored training I will gain expertise in analytical chemistry, manage large collaborations from inception to publication, and enhance my mentoring and teaching skills. These training objectives are in service of my long-term career goal of leading an independent research team focused on the synthesis, localization, and essential roles that Mtb cell envelope lipids and glycans play in bacterial survival and human disease. In my R00 phase I will apply for independent R01 and other funding, beginning a career with meaningful impact on the treatment, prevention, and diagnosis of human tuberculosis.

NIH Research Projects · FY 2026 · 2026-04

PROJECT SUMMARY Poly (ADP-ribose) polymerase inhibitors (PARPi’s) are a promising treatment for BRCA-associated breast cancer, exerting synthetic lethal effects in homologous recombination (HR) repair-deficient tumor cells, yet clinical results are not durable. Our team has demonstrated that PARPi-mediated DNA damage in tumors also activates cGAS/STING signaling resulting in immune infiltration, including T-cells. Although these immunomodulatory effects are critical for maximal efficacy of PARPi’s, to date, the addition of PD-1/L1 blockade has not improved PARPi efficacy in preclinical models or in clinical trials. Our recent preclinical work has demonstrated that PARP inhibition also results in infiltration of CSF-1R+ tumor-associated macrophages (TAMs) that carry a highly immunosuppressive phenotype. CSF-1R blockade depletes these TAMs and significantly augments PARPi efficacy in the K14CreBrcaf/f;Tp53f/f mouse model in a CD8+ T-cell-dependent manner. Based on these data, we recently activated a Phase 1 clinical trial, AXALAP, that combines the anti-CSF-1R antibody axatilimab and olaparib in patients with PARPi-naïve or sensitive metastatic BRCA-associated breast cancer. In Aim 1, these preclinical findings will be validated in tumor biopsies procured pre-treatment, after a 2-week lead- in of olaparib alone, and after combined olaparib plus axatilimab from patients enrolled in the AXALAP clinical trial. Bulk and single cell RNA sequencing and cyclic immunofluorescence (CyCIF) will be used to deeply characterize both the T-cell and TAM components of the tumor microenvironment in response to olaparib plus axatilimab. TAM diversity before and on-treatment will be explored based on transcriptionally defined unique subsets. The topology and spatial relationships of TAMs with T-cells will also be explored. In Aim 2, we will leverage our recent discovery that CSF-1R(neg)CD40+ TAMs are maintained in murine tumors after PARPi plus anti-CSF-1R therapy and are also present in human breast tumors treated with PARPi therapy. CD40 agonists have been shown to activate TAMs and dendritic cells (DCs) to induce potent anti-tumor T cell responses in both the preclinical and clinical setting. Therefore, CD40 agonism may enhance the activity of the PARPi plus anti- CSF-1R combination. Here, we will assess the effects of PARPi + anti-CSF-1R in combination with CD40 agonism on tumor growth in long term efficacy studies and on the TME using sophisticated immunophenotyping in Brca1-deficient mouse models. In Aim 3, we will characterize DNA repair-based and immunologic mechanisms of resistance in olaparib-resistant tumors. Combined CSF-1R blockade and PARP inhibition will be studied in our preclinical immunocompetent models of Brca1-deficient breast cancer that have been trained to be PARPi-resistant. Combined PARP inhibition and CSF-1R blockade will also be studied with CD40 agonism to activate anti-tumor macrophages and DCs to overcome PARPi resistance. Taken together, these studies will define the T-cell and TAM components of the TME in response to PARP inhibition in patient samples and develop the strategy of combined CSF-1R blockade and PARP inhibition in PARPi-sensitive and resistant tumors.

NIH Research Projects · FY 2026 · 2026-04

Modified Project Summary/Abstract Section T cells control immune response during vaccination, autoimmunity and infection based on their recognition of antigens. This R01 resubmission proposal studies lipid antigens for T cells by taking advantage of a sensitive lipidomics platform that can comprehensively detect lipids bound to CD1 antigen presenting molecules. By detecting hundreds of lipids bound to human CD1a, CD1b, CD1c and CD1d, we will determine the general patterns of lipid capture by cellular CD1 proteins. By separating CD1 proteins that do and do not bind to T cell receptors, lipidome-scale experiments will identify chemical patterns in natural lipids that lead to activation and blockade of T cell response. Based on a prior program that identified CD1a ligands that block T cell response in vivo, we will determine the structures of CD1b, CD1c and CD1d ligands that are inhibitors for T cells. Finally, optimized blockers will be tested ex vivo using polyclonal human T cells and CD1 tetramers, with an emphasis on CD1a and CD1c blockers. Overall, we seek to develop anti-inflammatory therapies that can act through specific blockade of human T cells. Unlike the MHC antigen presenting molecules, CD1 proteins are non-polymorphic, meaning that once the CD1-lipid binding patterns are described, they will apply to nearly all humans.

NIH Research Projects · FY 2026 · 2026-04

Project Summary Sepsis and septic shock are the leading causes of morbidity and mortality among critically ill patients in intensive care units (ICUs), leading to extraordinary healthcare costs annually. Numerous attempts have been made to identify more specific and effective therapeutic strategies and pharmacological agents for the treatment of sepsis. None has proven effective to date. Thus, there is still a critical need for targeted and effective therapy to reduce mortality from this disease. The objective of this study is to elucidate the role of ROS in modulating IL-1β bioactivity in sepsis. The hypothesis is that ROS-induced irreversible oxidation and its modulation by S-glutathionylation are key regulatory mechanisms controlling IL-1β bioactivity in sepsis. There is overwhelming evidence that ROS and oxidative stress play significant roles in the pathogenesis of sepsis-associated multiple organ failures. Antioxidants, such as N-acetylcysteine (NAC), due to their anti- inflammatory property, have been proposed for the treatment of sepsis. However, the results of the related clinical trials with septic patients have generally been incongruous and disappointing. It turns out that, besides their pro-inflammatory and pro-injury roles in the pathogenesis of sepsis, ROS can also elicit anti-inflammatory effects by inhibiting IL-1β bioactivity. NAC ameliorates oxidative stress and ROS-induced tissue damage. However, such treatment also augments IL-1β bioactivity and IL-1β-mediated pro-inflammatory responses, thereby contributing to the ineffectiveness and even adverse effect of NAC-based sepsis therapy. In this study, it is proposed that combined treatment with NAC and IL-1 receptor antagonist, anakinra, will effectively attenuate sepsis pathogenesis. The premise is strongly supported by the preliminary finding that NAC treatment decreased sepsis-induced mortality more effectively in IL1R1 deficient mice than in WT mice. In current study, the role of ROS-elicited oxidation in modulating IL-1β bioactivity will be further investigated in a murine model of polymicrobial sepsis induced by cecal ligation and puncture (CLP). First, the effects of co- treatment with NAC and anakinra on sepsis pathogenesis will be examined in detail (Aim 1). Mechanistically, to elucidate the role of IL-1β irreversible oxidation in sepsis, the levels of irreversibly oxidized (inactive) IL-1β in septic mice will be measured, and the impact of IL-1β irreversible oxidation on CLP-induced sepsis will be determined (Aim 2). Additionally, the roles of S-glutathionylation and Grx1 in regulating the irreversible oxidation and deactivation of IL-1β in sepsis will be investigated (Aim 3). Finally, the ROS-mediated regulation of IL-1β bioactivity and its correlation with disease severity will be explored using plasma samples from human sepsis patients (Aim 4). Together, the experiments proposed in these four specific aims will advance our understanding of the molecular control of IL-1β activity in general and provide insights into the mechanism of action of ROS-induced irreversible oxidation in controlling IL-1β bioactivity in sepsis, with the ultimate goal of solidifying the combinatorial treatment with NAC and anakinra as a novel therapeutic strategy for sepsis.

NIH Research Projects · FY 2026 · 2026-04

This proposal outlines a research and career development plan to develop a transposon-based gene drive to combat antimicrobial resistance (AMR). AMR is an existential threat to modern medicine and global public health, causing over 2.8 million infections and 35,000 deaths annually in the U.S. alone. E. coli ST131 exemplifies this threat as a globally disseminated pathogen driving the epidemic spread of AMR. Limited development of new antibiotics coupled with rapidly emerging resistance creates an urgent need for novel approaches to combat AMR in this pathogen. Dr. Basta will address this challenge by harnessing the prolific spread of plasmids and transposons, the primary mobile genetic elements mediating AMR, to develop a transposon-based gene drive (TnDrive) for rapid elimination of resistance genes from bacterial populations, focusing on E. coli ST131 as a model pathogen. The central hypothesis is that the natural gene drive properties of plasmids and transposons creates an ideal solution to AMR that can be deployed in both medical and ecological settings. In preliminary work, Dr. Basta engineered a TnDrive proof-of-concept by combining a self-transmissible plasmid with a programmable transposon targeting an aminoglycoside resistance gene. Starting at a frequency of 1:20,000, TnDrive spread to virtually all cells within one day and eliminated the targeted AMR gene from over 99% of the population. To optimize TnDrive, Dr. Basta will: (1) enhance transmission by identifying and circumventing bacterial defenses and metabolic barriers to plasmid spread and evaluating alternative plasmid chassis for improved environmental and in vivo efficacy; and (2) improve AMR elimination by multiplexing highly efficient targeting guides, establishing guide design rules for minimizing off-target effects, protecting against native transposon- mediated inactivation, and modulating host factors controlling transposition. The long-term goal is to develop TnDrive into a clinically and ecologically viable solution to combat AMR while advancing our understanding of transposon biology, plasmid transfer dynamics, and genome engineering. The candidate, David Basta, MD, PhD, is completing clinical pathology residency at Brigham and Women’s Hospital (BWH) and working with Dr. Matthew Waldor, a leading pathogen geneticist who has trained over 30 independent investigators. The project builds upon novel high-throughput functional genetic tools developed by Dr. Basta in the Waldor lab during his pathology residency. This work will be conducted at BWH, which provides exceptional training resources and access to a large collection of genetically characterized clinical isolates through its pathogen genomics program. Dr. Basta is supported by an advisory committee with expertise in transposon biology, AMR, bacterial genetics and genomics, gene drives, and animal models of infection. This career development award will enable Dr. Basta’s transition to independence as a physician-scientist integrating expertise in clinical pathology, molecular microbiology, genome engineering, and synthetic biology.

NIH Research Projects · FY 2026 · 2026-04

Project Summary Allergic diseases persist throughout a person’s lifetime, and the mechanisms underlying maintenance of the Th2 cells that drive these diseases are poorly understood. Chronic inflammation canonically drives T cell exhaustion in chronic infection and cancer, while Th2 cells conversely persist despite chronic inflammation We recently created a single-cell transcriptomic atlas from human tissues with allergic type 2 inflammation. A conserved set of Th2 cells was identified across tissues, including an aberrant memory-like population co-expressing TCF1 and LEF1, with evidence of chronic activation. This transcriptional profile mirrors the progenitor/stem-like CD8+ T cells that have been described in cancer and chronic infection, and we hypothesized that this Th2 cell population may serve as the reservoir that perpetuates Th2 responses. We named this cell the Th2 cell multipotent progenitor (Th2-MPP). Ex vivo functional studies established that these cells couple self-renewal with terminal differentiation in Th2, TFH- like, and Treg cells. We propose that aberrant cell-cell interactions and alarmin signaling networks in allergic tissues orchestrate an epigenetic state in Th2-MPP that enables self- renewal in the face of chronic antigen burden. The specific aims of this research strategy will investigate the epigenetic mechanisms and cell- cell interactions driving human Th2-MPP maintenance (Aim 1). Moreover, we will functionally interrogate candidate pathways through the development and characterization of a mouse model of chronic type 2 pulmonary inflammation, with temporal conditional genetic ablation of putative regulators of Th2 maintenance (Aim 2). These complimentary human and mouse immunology approaches will yield fundamental insights into the chromatin states of type 2 lymphocytes in barrier tissues and signaling mechanisms that drive this profile. Concurrently, the mechanistic studies of candidate pathways in mice will establish new avenues for the development of therapeutic modalities. This study combines mechanistic mouse models and primary human tissue analyses with cutting-edge single cell bioinformatics to provide the candidate new training in key elements of translational immunology. Through support from the co-mentors and Advisory Committee, the candidate will be well-positioned to transition to independence as an investigator, with the goal of identifying new therapeutic targets for the treatment of chronic inflammatory diseases.

NIH Research Projects · FY 2026 · 2026-04

Abstract The overarching goal of this project is to elucidate the role of biological sex in modulating the adverse metabolic effects of circadian misalignment, with the ultimate aim of designing sex-specific interventions to mitigate health risks associated with night shift work. Night shift work, prevalent in both male and female workforces, significantly increases the risk of metabolic disorders, including obesity and diabetes. A key contributor to these risks is circadian misalignment—misalignment between behavioral/environmental cycles and the body’s internal circadian system—which disrupts energy balance and impairs glucose tolerance. Despite established sex differences in human circadian physiology and metabolism, few experimental studies have explored whether these differences influence the metabolic consequences of circadian misalignment. Our pilot data from controlled laboratory studies provide proof-of-principle evidence of sex-specific effects. Acute circadian misalignment disrupted energy balance through distinct mechanisms: in women, it decreased satiety and increased hunger hormones, while in men, it elevated cravings for hyperpalatable foods. Preliminary findings also suggest that men are more susceptible to glucose intolerance under circadian misalignment compared to women. To build on this foundation, our proposed study addresses key limitations of prior work and systematically tests the following hypotheses: (Aim 1) Circadian misalignment leads to more adverse changes in appetite-regulating hormones in women than in men. (Aim 2) Circadian misalignment results in greater impairment of glucose control in men compared to women. (Aim 3A) Circadian misalignment increases homeostatic appetite in women and hedonic appetite (cravings for hyperpalatable foods) in men. (Exploratory Aim 3B) Circadian misalignment more adversely impacts food-related neurocognitive control (e.g., impulsivity and motivation) in men. (Exploratory Aim 3C) Circadian misalignment increases overall energy intake in women and promotes higher carbohydrate and saturated fat consumption in men. This study utilizes a balanced parallel group design (men vs. women), with each participant serving as their own control in a randomized, cross-over protocol (circadian alignment vs. circadian misalignment). Aims 1, 2, and 3A&B will be conducted under highly controlled laboratory conditions with energy-balanced diets. Aim 3C will evaluate ad libitum food intake on the final study day to assess potential implications without confounding other measurements. By uncovering sex-specific vulnerabilities to circadian misalignment, this project will provide novel insights into the physiological mechanisms driving shift work-related health risks. These findings hold broad implications for designing tailored shift work schedules, developing sex- specific interventions, and addressing sex disparities in clinical care, ultimately improving the health and well- being of millions of shift workers worldwide.