Massachusetts General Hospital

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY / ABSTRACT The airway epithelial surface is constantly exposed to the environment and serves as a barrier to many types of injury - toxins, infections, allergens, and other insults. The airway epithelial surface must regenerate after denuding injury. Both these aspects – barrier function and regeneration – can become dysregulated in diseases such as asthma, COPD, and bronchiectasis. We have discovered a new airway epithelial cell type, called the hillock, which survives various airway injuries and regenerates the epithelial surface to restore mucociliary transport. The overall hypothesis of this proposal is that the hillock plays a critical role in barrier function and regeneration in the airway and may become dysregulated in disease. Through single cell RNA sequencing and histological characterization, we find that the hillock highly expresses keratin 13 and desmoglein 3. These proteins are important in barrier function in of the skin, and we hypothesize that they similarly play a major role in the airway hillock. In Specific Aim 1 of this proposal, we will define the mechanism of hillock injury resistance. Using a hillock specific driver mouse we have developed, we will test this hypothesis by disrupting keratin 13 and desmosomes and assess for injury resistance by live/dead staining, proliferation, and electron microscopy. Hillocks also play an essential role in airway regeneration. We find when stimulated with retinoic acid, hillocks regenerate ciliated cells. In Specific Aim 2 of this proposal, we define the mechanism of ciliated cell differentiation from hillocks and restoration of mucociliary transport. It has been shown that different retinoic acid receptors may affect ciliogensis and oncogenesis. We hypothesize that retinoic acid receptor alpha and best leads differentiation of hillocks into ciliated cells which could then restore mucociliary transport. We will test this hypothesis in both mouse and human hillock explants. After performing luminal injury, we will regenerate the epithelia in the presence of retinoic acid receptor selective agonists and characterize ciliogenesis by immunocytochemistry, electron microscopy, cilia length measurements and functional assessment by micro-optical coherence tomography. This work is potentially paradigm shifting as it implicates a specific cell type, hillock cells, in disease pathology of multiple airway diseases and interrogates this cell type to uncover novel therapeutic targets. Dr. Shah will carry out this work under the mentorship of Dr. Jay Rajagopal, a leader in stem cell biology with an outstanding record of guiding young investigators to independence. Through this proposal, Dr. Shah will obtain training in (1) inducible genetic models, (2) developmental and stem cell biology, (3) explant models of disease, and (4) functional real-time imaging, which is only possible in the environment of the Rajagopal lab and collaborators at MGH. An advisory team of complementary and diverse scientists have been assembled to provide breadth and depth to the training plan. Hands-on training will be supplemented with coursework in stem cell biology and advanced imaging, necessary for Dr. Shah to become an independent R01 funded investigator.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Chronic obstructive pulmonary disease (COPD) is the leading respiratory cause of death worldwide, and by 2050, sub-Saharan Africa (sSA) will bear the greatest COPD burden despite the lowest smoking rates in the world. HIV is an important yet underappreciated driver of COPD risk that leads to more aggressive and fatal disease, particularly among women, for reasons that have not been fully elucidated. The intersection between HIV, sex, and COPD risk is particularly important in sSA, where most of the global population with HIV lives, over half of whom are women and girls. Consequently, two major public health priorities include understanding the mechanisms of COPD in women with HIV in sSA and developing methods to identify at-risk individuals and interventions to prevent or reduce its impact on their health. To accomplish these priorities and respond to the growing epidemic, data are needed in this population prior to the development of COPD at potentially intervenable timepoints. Unfortunately, longitudinal lung health data prior to COPD and mechanistic data to elucidate pathobiology and identify at-risk populations are rarely available in sSA. This proposal is led by experts in adult and adolescent lung disease epidemiology, quantitative chest imaging analytics, proteomic biomarker science, and risk prediction scores in sSA. In this proposal, we will build upon our preliminary data that suggest that women with HIV have particularly elevated risk of HIV-associated lung disease by establishing the Uganda Lung Health Study, a cohort of 800 adolescents and adults that is evenly balanced by sex and HIV and spans the entire adult lifespan from 16 to 90 years of age. We will follow these 800 individuals with three annual study visits where we will collect post-bronchodilator spirometry, study questionnaires, blood samples, and non- contrast chest CT imaging to accomplish the following aims: Aim 1: To identify trajectories of lung dysfunction across the lifespan and their key risk factors in adolescent girls and women with HIV in Uganda. We will estimate lung function trajectories via group-based trajectory modeling using 2,400 spirometry measures from 16 to 90 years old. Our secondary outcomes are categorical lung dysfunction and parenchymal lung disease. Our primary predictors are HIV and sex, and we will also consider demographic, environmental, clinical, and HIV-specific factors. We will identify key risk factors through clinical prediction scores and clinical prediction algorithms. Aim 2: To identify candidate biological pathways that predict lung dysfunction among adolescent girls and women with HIV in Uganda. We will assess biological pathways using proteomic signatures, and will compare signatures predictive of abnormal lung function trajectory and parenchymal lung disease by sex and HIV serostatus, with the goal of motivating future work to develop HIV- and/or sex-specific interventions. Our work will provide insights into potentially targetable risks and biological pathways in Uganda and will guide mechanistic and intervention studies to preserve lung health among PWH, which is directly responsive to NHLBI, Office of AIDS Research, and Office of Research on Women’s Health research priorities.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Increasing numbers of adolescents are presenting for mental health care with symptoms consistent with such serious mental illnesses (SMI) as schizophrenia and depression. Many SMI exhibit pluripotent risk profiles during the prodromal phase, underscoring the need to develop novel interventions that target transdiagnostic mechanisms and ameliorate distressing symptoms in adolescents at risk for SMI. Repetitive negative thinking (RNT), maladaptive, repetitive, intrusive, unproductive thought patterns, are associated with a broad range of SMI, and with symptom severity, treatment non-response and relapse. At the neural level, RNT is characterized by elevated functional connectivity within the default mode network (DMN), and similarly, prior research has consistently demonstrated patterns of DMN hyperconnectivity in SMI. Interestingly, mindfulness meditation, which trains attentional focus to the present moment, reduces RNT. Adolescents can apply mindfulness practices to decrease perceived stress, increase sustained attention, and suppress DMN activity. Although mindfulness has profound mental health benefits, for some, mindfulness alone may not be sufficient to mitigate RNT because RNT itself and other mental health symptoms may impede progress in successfully acquiring and utilizing mindfulness strategies. To directly address this challenge, we propose using real-time fMRI neurofeedback to enhance the acquisition and utilization of mindfulness skills to better target DMN hyperconnectivity and transdiagnostic RNT symptoms. In our previous research, we developed a novel, 15-minute mindfulness-based, real-time neurofeedback (mbNF) paradigm whereby people observe a visual display of their brain activity and practice mindfulness to volitionally reduce DMN activation. In the R61 phase, 50 at-risk adolescents and 50 healthy control adolescents (ages 13-16) will receive a localizer resting state scan. We will first confirm at-risk adolescents exhibit higher DMN connectivity than controls (Go criterion 1). At-risk adolescents will be further randomized to receive a 15- or 30-minute ‘dose’ of mbNF (n=25/group). We predict reduced DMN connectivity across doses (target engagement; Go criterion 2). We also predict at-risk adolescents will show greater reduction of DMN hyperconnectivity after 30 minutes of mbNF compared to 15 minutes. In the R33 phase, a new sample of 90 help seeking adolescents (ages 13-16) with RNT will participate in a double-blind randomized controlled trial of two sessions of either active or sham mbNF (n=45/group) added to mindfulness training. We will test–using clinician-administered instruments and self-reports –whether two sessions of active mbNF compared to two sessions of sham mbNF contributes to a greater reduction in RNT (primary outcome) and psychiatric symptom severity (secondary outcomes) across the 1-month, 3-month and 1-year assessments. As a whole, mbNF is directly in line with precision medicine initiatives, and if successful, could revolutionize clinical care for at-risk adolescents.

NIH Research Projects · FY 2025 · 2025-09

Project Summary/Abstract Systemic lupus erythematosus (SLE) is a chronic autoimmune disease associated with excess morbidity and premature mortality. Up to half of all patients with SLE will develop lupus nephritis, which also contributes to heightened risks of cardiovascular (CV) events and end-stage kidney disease (ESKD). These adverse outcomes are driven by a combination of uncontrolled lupus disease activity, chronic tissue damage including proteinuric nephropathy, and glucocorticoid toxicity, which exacerbates conventional CV risk factors. A major unmet need in lupus care is to improve kidney and CV outcomes, and novel treatment approaches are needed. Glucagon-like peptide-1 receptor agonists (GLP-1RAs) are a class of anti-obesity and hypoglycemic agents with pleiotropic cardioprotective, nephroprotective, and anti-inflammatory effects. These medications have been found to prevent major adverse cardiac events (MACE) and reduce kidney disease progression in non- SLE populations, including those with overweight/obesity and/or type 2 diabetes. Our central hypothesis is that GLP-1RA medications will have similar to greater benefits in patients with SLE. The goal of this proposal is to investigate the potential role for GLP-1RA to improve outcomes for patients with SLE and lupus nephritis. In Aim 1, we will leverage a large SLE cohort in an administrative healthcare database to emulate target trials to test the hypothesis that GLP-1RA use versus no GLP-1RA use will reduce the risk of MACE and kidney disease progression, including new-onset ESKD, among patients with SLE and overweight/obesity. We will also utilize an electronic health record (EHR)-based SLE cohort from multiple academic and community hospitals to test the hypothesis that GLP-1RA use among patients with SLE will be associated with improvements in conventional CV risk factors (including blood pressure, cholesterol, and body weight/body mass index) and a reduction in proteinuria in the subgroup of patients with lupus nephritis. In Aim 2, we will use the same EHR-based SLE cohort to test the hypothesis that GLP-1RA use will be associated with a lower risk of lupus flares and reduced biomarkers of disease activity. The anticipated findings will provide critical evidence for the potential benefits of GLP-1RAs in treatment of patients with lupus. If successful, the anticipated findings will lead to a new treatment paradigm and improvements in SLE care.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT: Gout is the most common inflammatory arthritis worldwide caused by the deposition of monosodium urate (MSU) crystals. The disease burden of gout is high and continues to rise, with >12 million US adults being affected by gout. In addition to causing recurrent and excruciating flares of arthritis, gout is independently associated with cardiovascular disease (CVD) and contributes to premature mortality. This premature mortality among patients with gout persists even after adjusting for atherosclerotic CVD risk factors, suggesting that gout-specific mechanisms contribute to premature mortality among patients with gout. One hypothesized gout-specific mechanism is the direct deposition of MSU crystals in the vasculature, which can activate the NLRP3 inflammasome and interleukin-1ß pathway locally; this same pathway is heavily implicated in the pathogenesis of atherosclerotic CVD in response to cholesterol crystals. Multiple pathology and optical coherence tomography studies have demonstrated evidence of microscopic MSU crystals in the vasculature. Subsequently, there has been surging interest in the use of dual-energy computed tomography (DECT) for the non-invasive identification of macroscopic MSU crystals in the vasculature. Building on this interest, and our recent multi-center cross-sectional coronary DECT study (CORODECT) in which we found that patients with gout had 2.7-fold higher odds of having DECT findings consistent with vascular MSU deposition compared to controls, we propose to conduct the first-ever longitudinal study to determine if effective urate-lowering therapy leads to the reduction in these vascular MSU lesions. This imaging study will be ancillary to the multi-center Treat-to-Target Serum Urate versus Treat-to- Avoid Symptoms in Gout: A Randomized Controlled Trial (TRUST; U01 AR080985). In this time-sensitive proposal, we will obtain DECT scans of the coronary arteries and bilateral feet at baseline and month 18 among TRUST participants (all with serum urate ≥6 mg/dL) randomized to treat-to-target serum urate (TTT-SU, intensive urate-lowering) versus treat-to-avoid-symptoms (TTASx, symptom-focused management). Aim 1 will assess whether TTT-SU is associated with greater reductions in vascular MSU deposit volumes and numbers than TTASx over 18 months (we hypothesize TTT-SU will be more effective), whilst Aim 2 will assess the correlations between baseline MSU volumes in the feet and vasculature, and correlations in changes in MSU volumes at these sites. TRUST provides an unparalleled opportunity to conduct a longitudinal DECT study among patients with well-phenotyped gout recruited from primary care practices, taking full advantage of the randomization of the intervention, to fill a critical evidence gap in our understanding of the role of vascular MSU deposits as a gout-specific mechanism contributing to CVD and premature mortality. The anticipated findings will provide compelling rationale and immediately actionable evidence for the use of ULT as a cardioprotective intervention among patients with gout.

NIH Research Projects · FY 2025 · 2025-09

Project Summary/Abstract Over 5 million Americans aged 65 and older identify as Hispanic or Latino (hereafter Latino), nearly two-thirds of whom speak Spanish at home. Compared to non-Latino White older adults, Latino older adults are also more likely to have a serious illness, a health condition that carries a high risk of mortality and negatively impacts a person’s daily function or quality of life. Despite high rates of serious illness, Latino older adults have less engagement in serious illness communication (SIC) compared to non-Latino counterparts. SIC is a process of structured, emotionally supportive conversations about prognosis, values, and priorities to inform care decisions. High-quality SIC is associated with improved quality of life, care aligned with patient preferences, and enhanced clinician satisfaction, while poor communication is linked to emotional distress and lower-quality care. Multiple tools to teach SIC exist, but early research examining their acceptability and impact was conducted primarily in White, English-speaking populations whose communication needs may not reflect those of more diverse populations. Critical care gaps remain for SIC with Latino older adults: (a) clinicians cite barriers to initiating SIC with diverse patients, (b) interpreters report that interpreted SIC conversations usually go well only half of the time, and (c) clinicians receive little guidance on how to navigate SIC when the patient and clinician have language discordance, which is more common in older adults. To fill these gaps, the applicant has conducted initial research on the SIC experiences of hospitalized Latino Spanish-speaking patients and inpatient palliative care clinicians. However, SIC differs between care settings and requires different skills in community-based (clinic and home-based) settings: conversations about urgent, real-time decisions in the hospital setting differ from conversations outside the hospital where clinicians support patients in care planning over time with more time for rapport-building and addressing coping. The objective of this proposal is to explore SIC needs and experiences of Latino Spanish-speaking older adults engaging in community-based palliative care from the perspective of patients, palliative care clinicians, and professional medical interpreters. This proposal investigates two specific aims: (1) to explore the SIC needs and experiences of Latino Spanish-speaking older adults in community-based palliative care settings (half who had a language-concordant visit and half who had a language-discordant visit), and (2) to explore monolingual and bilingual palliative care clinicians and professional medical interpreters’ perspectives on the unique needs, experiences, barriers, and facilitators with conducting SIC with Latino Spanish-speaking older adults in community-based palliative care settings. This work will inform future clinician- and interpreter-facing SIC skill development interventions and improve serious illness communication and care for Latino Spanish-speaking older adults. This work will lay the groundwork for expanding SIC skill development interventions to different clinicians (e.g., primary care and oncology) who support Latino Spanish-speaking older adults.

NIH Research Projects · FY 2025 · 2025-09

The overarching goal of this proposal is to improve the quality of life of lymphoma survivors. To achieve this goal, we propose to test the efficacy of a virtual, resilience group program (SMART3RP-lymphoma) for lymphoma survivors delivered during early posttreatment survivorship (0-2 years after active lymphoma treatment [surgery, chemotherapy/immunotherapy/radiation] with curative intent). Given the nature of their disease (i.e., a cancer of the immune system) and the use of intensive, multi-agent therapies, lymphoma survivors experience a broad and persistent array of psychosocial challenges after treatment that complicate their adjustment and negatively impact their quality of life.1-8 As a treatable cancer that commonly presents in younger patients (20% occur in young adults, aged 18-39), these survivors often face longer lifespans grappling with the impact of lymphoma and its treatment.3,4 Given their potential to live for decades posttreatment, how lymphoma survivors adjust to the aftermath of their disease can have a profound impact on their health and wellbeing. Unfortunately, due to perceptions of lymphoma as a “good cancer” and a general lack of attention to the posttreatment period, the needs of these survivors are often overlooked.8,11 The end result is a population left at elevated risk for experiencing significant and enduring declines in their quality of life. To address these evidence gaps, we aim to test the effects of an evidence-based, multicomponent, resilience intervention (SMART3RP-Lymphoma)7 in a randomized controlled trial with early post-treatment lymphoma survivors (n=254) from 3 NCI-designated cancer centers. Specifically, this study aims to test the efficacy of SMART3RP- Lymphoma, compared to a Health Education Program in improving quality of life (primary outcome) as well as anxiety, depression and physical functioning (secondary outcomes). We will also explore how changes in resilience impact quality of life – a critical yet understudied priority in cancer survivorship. Further, in subgroup analyses, we will examine how SMART3RP-lymphoma impacts patient subgroups. Together with stakeholder and participant qualitative data, this trial will provide critical information to guide next steps in evaluating the cost, implementation, and sustainability of SMART3RP-lymphoma across settings. The proposed research represents a substantive departure from the status quo by offering a new post-treatment psychosocial care paradigm that emphasizes strengths-based approaches to improve quality of life in survivors. If effective, this model can be adapted and tested with other survivor cohorts. This work thus has the potential to have a significant impact on clinical practice and public health.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Obesity affects more than 40% of the United States population and is a major risk factor for acute hypoxemic respiratory failure (AHRF). AHRF patients with obesity have more ventilator-related morbidity and may be less responsive to standard respiratory care treatments, such as prone position and positive end expiratory pressure (PEEP) titration, compared to patients of normal body weight. Excess truncal adiposity in obesity elevates intrathoracic pressure (ITP) and compresses lung tissue. Our prior research has shown that obesity also modifies ventilator-induced lung stretch. In addition to the mass of adiposity, published and preliminary data suggest that the distribution of adiposity influences ITP and may affect the magnitude and pattern of lung collapse. Imaging the topographic effects of truncal adiposity on lung tissue may therefore 1) provide insight into AHRF severity and treatment response in obesity and 2) identify viable tools that can quantify the effects of truncal loading at bedside. Our research group has extensive expertise in applying novel computed tomography (CT) image analysis involving registration and artificial intelligence-based segmentation to quantify lung collapse and injury progression in AHRF. In this proposal, we will use CT to test our hypothesis that the magnitude and distribution of truncal adiposity governs lung collapse and severity of AHRF in obesity. Using CT as reference, we will further evaluate how measurements of ITP (esophageal manometry), global respiratory mechanics (forced oscillation technique), and regional ventilation (electrical impedance tomography) can detect the effects of truncal loading on lung collapse at bedside. To achieve the aims of this proposal, we have developed a ventilated swine model of AHRF and a robust clinical infrastructure to monitor patients with AHRF in our hospital. We will first measure in our animal model the effect of load distribution on imaging, biological, and physiological markers of AHRF progression during prolonged mechanical ventilation for 24 hours. We will then evaluate the effects of prone positioning and PEEP titration on the regional inflation characteristics and radiological abnormalities using sequential imaging in conditions of truncal loading. Lastly, in ventilated patients with AHRF and in healthy volunteers with and without obesity, we will measure associations between thoracic adiposity, ITP, and lung collapse in supine and prone position. Data obtained from our proposal will advance our understanding of obesity in AHRF, informing individualized clinical management to assist in the early identification and treatment based on the morphological and biomechanical characteristics of each patient.

NIH Research Projects · FY 2025 · 2025-09

PROJECT ABSTRACT/SUMMARY Chronic obstructive pulmonary disease (COPD) and asthma are the two most common chronic respiratory diseases, with COPD as the third leading cause of death globally and asthma as the most common chronic disease of childhood. They are treatable, heritable, strongly affected by the environment (e.g. smoking, pollution), and exhibit some of the largest disparities in prevalence and survival among populations. Genetic and multi-omic technologies provide powerful opportunities to identify both shared and distinct heritable risk factors among asthma and COPD, connect those genetic variants with function, and delineate relationships between genetic, environment, and disease risk using multi-omic molecular assays. However, most genetic studies of lung diseases and comorbid traits to date have focused narrowly on one or few traits at a time without modeling the dense correlation between risk factors and have been vastly Eurocentric. This is problematic, as minority communities who face a higher burden of respiratory diseases disproportionately experience adverse exposures that can be challenging to study because they are often imprecisely measured, and the level of exposure that is biologically meaningful for disease onset and progression is unclear. In this R01 application, we will integrate multi-trait, multi-ancestry, and multi-omic data from massive biobanks. We will bring together genetic and phenotypic data across the All of Us Research Program, Million Veterans Program, UK Biobank, FinnGen, and other global biobanks, totaling more than 2 million participants, including approximately 240,000 COPD cases, 260,000 asthma cases, and 60,000 proteomic profiles. In Aim 1, we will conduct powerful genetic studies of lung diseases and comorbidities, model genetic correlations among related traits and ancestries to maximize genomic discovery, refine putative causal variants, and predict genetic risk accurately across populations. In Aim 2, we will connect asthma and COPD genetic risk loci to function by using their pleiotropic relationships to dissect them into sets with similar effects, then test for enrichments and colocalizations among these sets of variants with multi-omic molecular assays to elucidate their molecular mechanisms. Lastly, in Aim 3, we will bridge the gap between inherited and environmental influences to better understand phenotypic heterogeneity in asthma and COPD by leveraging proteomic and metabolomic profiles to develop scores that predict incident disease and well-known risk factors. This R01 application will deliver accurate and reproducible genetic, environmental, and multi-omic molecular predictors of asthma and COPD that will identify causal mechanisms and modifiable risk factors. Quantifying the relative importance of risk factors as proposed here is critical for advancing clinical models, nominating targeted interventions to improve public health, and ultimately reducing vast health disparities in lung disease.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT The goals of this Mentored Patient-Oriented Research Career Development Award are two-fold: (1) evaluate the feasibility, acceptability, and preliminary clinical utility of measuring dyadic relational processes during early recovery from alcohol use disorder (AUD) using ecological momentary assessment (EMA) to identify novel proximal dynamics influencing early recovery outcomes and inform intervention designs targeting relational mechanisms in both patients and romantic partners, and (2) establish the principal investigator as an independent researcher at Massachusetts General Hospital-Harvard Medical School. The specific aims of this research will be accomplished through an innovative study leveraging the strengths of dyadic ecological momentary assessment and qualitative interviewing to collect interpersonal dynamics data within couples during initial recovery. This research will serve as the methodological foundation for future studies illuminating relational dynamics between people in recovery and close supportive others to understand how the proximal social context impacts recovery outcomes. Further, this innovative study will inform interpersonal treatment approaches and will characterize the cycle of shame and intentional withholding of information in initial recovery. Aims of the principal investigator’s career development and training plan include: (1) Extend expertise in etiology of addiction to recovery science; (2) Cultivate independent skills in social processes, couples’ interactions, and dyadic momentary data collection and data analysis; (3) Develop expertise in qualitative research; and (4) Foster transition from mentored postdoctoral training to independent translational clinical scientist. These goals will be achieved through an interactive training plan comprised of mentorship, formal coursework, seminars, conferences, and manuscript preparation. Knowledge gained via the training plan will be augmented and enhanced through research endeavors planned throughout the funding period. Drs. John Kelly, Jaye Derrick, Aidan Wright, and Christina Psaros will serve as mentors on this award, and will provide expertise in couples interactions, dyadic processes and dyadic ecological momentary assessment, dyadic analytic approaches, qualitative measurement, and addiction recovery science. Massachusetts General Hospital-Harvard Medical School provides an exceptional environment in which to conduct research and launch a career as an independent scientist engaged in alcohol use disorder treatment development. By the end of the 5-year award period, the goals are to validate an interpersonally-focused ecological momentary assessment battery to be used in a larger study of early recovery with key supports beyond romantic partners in the context of an R01, to accumulate initial content necessary for an R34 submission to create just-in-time interventions for couples experiencing an initial recovery attempt, and for the PI to be established as an independent investigator. This award is consistent with NIAAA’s 2024-2028 strategic plan to explore the behavioral, environmental, and social mechanisms that impact recovery outcomes to advance research on recovery from AUD (Goal 4, Objective 3).

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY: Maternal mental health disorders are the leading identifiable cause of childbirth-related maternal death in the U.S. and a significant contributor to maternal morbidity and health disparities. Among these disorders, childbirth-related posttraumatic stress disorder (CB-PTSD) remains an underdiagnosed and debilitating condition. CB-PTSD affects approximately 240,000 American women annually who experience traumatic childbirth. It is marked by symptoms that emerge soon after delivery and are triggered by reminders of the traumatic birth, with the child often becoming an unfortunate symptom enhancer. This dynamic can impair mother-infant bonding during critical developmental stages. The urgency of addressing CB-PTSD is heightened by its intersection with life-threatening complications during childbirth and severe maternal morbidity (SMM), which are highest in the U.S. among Western countries and disproportionately affect minoritized women. Hence, the need to understand and mitigate the psychological aftermath of medically complicated childbirth. Despite recommendations for universal maternal mental health screening in U.S. hospitals, no tools currently exist to identify women at risk for CB-PTSD before symptoms fully develop. This proposal seeks to address this critical gap by developing an innovative, multi-modal screening tool for early detection of CB-PTSD. Leveraging the unique opportunity afforded by childbirth trauma, a clearly defined event with early symptom onset, we propose a prospective, longitudinal study to assess women with complicated, potentially traumatic deliveries. Beginning in the first postpartum days, we will collect oral unstructured short childbirth narratives at the bedside depicting the subjective birth experience. We will measure psychophysiological responses during recollection of the childbirth trauma using validated methods to assess objective emotional reactivity. Obstetrical data, a proxy of the magnitude of the traumatic childbirth event, will be obtained from electronic medical records. Participants will be followed at multiple postpartum time points (Week 2, and Months 1, 3, 6) to assess acute and chronic CB-PTSD via validated psychometric tools and diagnostics. In our prior work, the potential for artificial intelligence (AI) models to detect CB-PTSD via childbirth narratives is demonstrated. We will expand this innovative line of research in this application. Using state-of-the-art AI and large language models (LLMs), we will analyze these narratives and develop a machine learning framework that integrates subjective accounts, objective physiological markers, and obstetrical indicators to predict CB-PTSD risk. The anticipated outcomes include a cost-effective, scalable screening tool to identify CB-PTSD risk early, enabling timely interventions and setting a new standard for trauma-focused postpartum mental health care. This project addresses an unmet clinical need, aligning with national priorities to reduce adverse maternal and child outcomes and to mitigate racial disparities in maternal health.

NIH Research Projects · FY 2025 · 2025-09

Project Summary Chronic kidney disease (CKD) affects up to 13.4% of the global population, imposing significant economic burdens on healthcare systems. While kidney tubular cells can self-repair, severe or repetitive damage may lead to degenerative fibrosis, revealing critical gaps in understanding adaptive cellular repair mechanisms. Our recent work has identified unique molecular profiles in repairing tubules, indicating that FANCD2, a key protein in the Fanconi anemia pathway and crucial for interstrand crosslink (ICL) repair, may regulate adaptive repair. Despite extensive studies on FANCD2 in DNA repair and cancer, its role in the kidneys and CKD is poorly understood. No studies have reported the kidney phenotype of FANCD2 knockout (KO) mice; however, data from the International Mouse Phenotypic Consortium (IMPC) suggest kidney degenerative changes in heterozygous KO males. Furthermore, clinical studies indicate an association between Fanconi anemia and CKD, emphasizing the relevance of this research. Our investigation revealed a unique pan-nuclear distribution of FANCD2 in proximal tubules, distinct from typical DNA repair foci. Additionally, chromatin immunoprecipitation sequencing (ChIP-seq) analysis showed FANCD2 binding to the promoters of multiple genes, including those involved in kidney development. In cisplatin-induced acute kidney injury (AKI) mouse models, we observed protective effects of inhibiting FANCD2's canonical pathway. These findings suggest FANCD2 has multifaceted roles beyond its ICL repair, significantly impacting kidney development and repair through non-canonical pathways. Specific Aim 1 is to define the non-canonical role of FANCD2 as a transcriptional regulator in nephron epithelia. We will use diverse models, including conditional FANCD2 KO mice, transgenic human kidney organoids, and patient samples, employing techniques such as immunohistochemistry, 3D imaging, western blotting, qPCR, RNA-seq, and ChIP-seq. These integrative analyses will assess FANCD2 chromatin binding and transcriptional regulation. Specific Aim 2 is to investigate FANCD2’s roles in AKI and CKD through both canonical and non-canonical pathways using in vivo and in vitro models. We will employ genetic and pharmacological approaches to uncover these roles. Our evaluation will include nephron epithelial integrity, fibrosis, DNA damage and repair, and other processes through single cell analyses, providing insights into FANCD2’s pathways as a potential therapeutic target. These aims utilize diverse models to rigorously test our hypothesis and provide evidence for potential CKD interventions. Our comprehensive approach aims to enhance understanding of FANCD2-mediated kidney repair mechanisms. Unraveling the non-canonical roles of FANCD2 in CKD may lead to targeted therapies that halt fibrotic progression and improve renal function in CKD patients.

NIH Research Projects · FY 2025 · 2025-09

Project Summary Interferon (IFN) signaling plays an important role in the host defense against invading pathogens. Activation of this innate immune response can be protective or detrimental for the host, depending on the stimulating agent. Major IFN-producing pathways, like the STING pathway, have been historically studied in the context of viral and bacterial infections. However, a recent transcriptomic study revealed upregulation of type I IFN pathway components in response to Candida albicans (Ca) infection, suggesting a significant role for IFN pathways in fungal pathogenesis as well. My preliminary data revealed that deletion of essential STING pathway components improved host survival following a Ca infection. Furthermore, I demonstrated that Ca DNA packaged in extracellular vesicles (EVs) triggers the STING pathway causing a lethal and hyperinflammatory response that clears the fungal infection. Still, the questions of pathway regulation, functional consequence, and broad application to other fungal organisms remain unanswered. By completing this mentored career development award, I will gain invaluable training in EV isolation and characterization, Nanostring analysis, flow-based analytics, and bioinformatics while also fine-tuning my skills with confocal microscopy and animal work. During the mentored phase of this award, I will optimize EV and EV DNA preparation from various fungal pathogens in the Vyas laboratory, I will design and validate any knockout cell lines or visualization needed to complete these aims (including Nanostring and RNA sequencing), and I will attend bioinformatics courses and career development seminars relevant to my future. In the independent phase I will focus on the animal survival studies (including breeding) outlined in Aim 1 and dedicate time to writing manuscripts and grants for my independent research. I will also use the training I received during my K99 phase in flow-based immunophenotyping and multiplex ELISAs as well as sequencing analysis to complete the experiments outlined in this grant. This proposal will continue to uncover the mechanism of STING-dependent type I IFN induction in response to clinically relevant fungal pathogens. In Aim 1, I will identify the functional significance for fungal EV induction of a specific ISG, viperin. In Aim 2, I will elucidate the regulatory mechanisms involved in STING pathway activation by fungal EVs and fungal DNA by exploring the role of an exonuclease (Trex1) known to regulate this pathway. Finally, in Aim 3, I will assess the extent to which fungal EVs activate the STING pathway. These findings will elucidate the functional consequence of this innate immune signaling pathway activation by fungal pathogens and uncover a new role for fungal EVs in host cell reprogramming.

NIH Research Projects · FY 2025 · 2025-09

Abstract The success of COVID-19 mRNA vaccines has showcased the effectiveness of this technology in rapid design, manufacturing, and adaptation to emerging viral threats. However, several imitations remain, including their instability requiring cold-chain storage that limits their global distribution, the necessity for two doses for optimal efficacy, limited immunity to closely matched viral strains, and quick-waning immunity following the initial two doses. These limitations can be potentially addressed by potent adjuvants. However, traditional adjuvants, co-delivered with vaccines to enhance antigen-presenting cell (APC) recruitment and functionality, are unsuitable for mRNA vaccines, because the co-delivered adjuvant provokes innate immune responses that interfere with mRNA translation, reducing its immunogenicity. Our recent investigation demonstrated that mRNA vaccines, particularly when delivered intradermally, could be substantially augmented by adjuvant delivery at different times and locations relative to mRNA vaccination, effectively circumventing the interference with mRNA transcription and translation. The current proposal aims to determine whether intranasal (IN) or intradermal (ID) delivery of a newly developed adjuvant can expand the breadth and longevity of trivalent influenza (flu) mRNA vaccine administered via a microneedle array patch (MNP). Specifically, we will construct a trivalent self-amplifying mRNA (saRNA) flu vaccine, encapsulated within liposomes free of cationic or ionizable lipids using our proprietary technology, and then loaded into our caved MNP after lyophilization. The antigen expression, immunogenicity, and efficacy of this saRNA flu vaccine will be optimized and evaluated in mice. Aim 2 will define the optimal time window of IN or ID adjuvant delivery following MNP-saRNA- flu immunization. Antigens-specific T-cell responses and broad neutralization antibodies across homosubtypic and heterosubtypic flu strains will be assessed and compared with or without the adjuvant and between IN and ID adjuvant delivery. The longevity of the specific immune responses will be also monitored over several months. Aim 3 will validate the efficacy of MNP immunization, alongside delayed adjuvant delivery, against various flu viruses in young and aging mice. If successful, the innovative strategy can be extended to other respiratory viral vaccines, such as those for COVID-19, respiratory syncytial viruses, etc., broadening the breadth and efficacy of various RNA-based vaccines delivered parenterally.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY This project aims to develop machine learning and artificial intelligence tools to enhance prediction of disease manifestation in individuals with pathogenic genetic variants. We will focus on Alzheimer's disease and cardiomyopathy, leveraging diverse data from large biobanks and healthcare systems to create a more comprehensive and nuanced understanding of how genetic variants interact with other factors to influence disease onset and progression. Our interdisciplinary team will create an ethical framework to guide tool development and implementation, integrating genomic, clinical, imaging, and other data types into a common model. We will build and validate tools to predict disease onset, progression, and treatment response. We will then perform rigorous cross-validation to assess the generalizability of these tools across different datasets and clinical settings, ensuring their robustness and applicability in diverse healthcare contexts. Using a combination of advanced AI and interpretable machine learning methods, we aim to identify novel biomarkers for disease that could be more readily translated into clinical practice. Integrated with the technical development, we will conduct research on the ethical, legal, and social implications (ELSI) of incorporating these tools into genomic medicine. The results of these ELSI research projects will be further integrated into our ethical framework to inform the development of the optimal AI tools. This project will advance the field by improving risk stratification for variant carriers and identifying factors influencing disease beyond genetics alone. We aim to disseminate validated tools, best practices, and lessons learned to the broader research community, potentially transforming how genetic risk is assessed and managed in clinical practice. By combining cutting-edge computational methods with careful ethical consideration, we hope to create resources that can significantly enhance patient care and our understanding of complex genetic diseases. Ultimately, this work will lead to more targeted prevention strategies, personalized treatment plans, and improved outcomes for individuals at risk of Alzheimer's disease, cardiomyopathy, and other genetically influenced conditions.

NIH Research Projects · FY 2025 · 2025-09

Project Summary Allogeneic cell therapies offer transformative potential in clinical medicine, providing universal treatments with donor-derived immune cells and mitigating the variability of autogenic therapies. Among immune cell types, natural killer (NK) cells and neutrophils stand out for their therapeutic potential. NK cells can effectively target tumors and virally infected cells without patient-specific matching, while neutrophils, which do not require ABO matching, show promise in treating infectious diseases and inflammatory conditions. However, widespread adoption of allogeneic therapies is limited by challenges in cell cryopreservation, as current methods often compromise cell viability and function due to osmotic stress and toxicity during CPA loading. To address these limitations, we propose veloporation, a novel mechanoporation technique leveraging viscoelastic stretching to enable rapid, high-throughput CPA loading while minimizing toxicity. Unlike traditional CPA-loading methods, which expose cells to two primary injury mechanisms—molecular toxicity from prolonged CPA exposure and osmotic stress from excessive volumetric excursions—veloporation is the first approach to load CPAs almost instantaneously without altering cell volume. By alleviating both injury mechanisms simultaneously, veloporation preserves immune cell phenotype and functionality post-thaw, offering a transformative solution to the critical barrier of effective cell storage. The goal of this project is to develop and validate veloporation for CPA loading (velCPA) into immune cells, with a focus on NK cells and neutrophils for use in adoptive cell therapy (ACT). Aim 1 develops and optimizes the veloporation device for small immune cells. Aim 2 evaluates the ability of veloporation to load membrane-permeable and membrane-impermeable CPAs, minimizing toxicity and maximizing cell viability compared to traditional methods. Aim 3 assesses the post-thaw functionality of veloporated cells, including viability, immune activity, and potential for CPA co-delivery with therapeutic agents. By advancing cryopreservation techniques, this project addresses a critical barrier to the scalability of allogeneic cell therapies. It has the potential to enable the development of stable, off-the-shelf immune cell products for diverse clinical applications. While here we focus on NK cells and neutrophils, this technology could be adapted for the cryopreservation of other cell types used in regenerative medicine and immunotherapy, such as T cells or stem cells. The versatility and scalability of veloporation make it a promising platform technology with far-reaching implications for cellular therapies and biobanking.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY CAR T cell therapy for solid tumors is hindered by a lack of tumor-specific antigens that are safe to target and homogenously expressed throughout the tumor, difficulty infiltrating the tumor due to dense tumor stroma, and suppression of CAR T cell function by the tumor microenvironment (TME). We plan to address these issues using novel meso-FAP CAR-TEAM cells that simultaneously target mesothelin, a solid tumor antigen that has already been proven safe to target in patients, and cancer-associated fibroblasts (CAFs), which inhibit T cell infiltration and suppress T cell function in the TME. The CAFs are targeted with T cell-engaging antibody molecules (TEAMs) secreted from the CAR T cells that bind to CD3 and fibroblast activation protein (FAP), which is highly expressed on CAFs. The TEAM allows for CAF elimination by CAR and non-CAR T cells in the tumor. We have already demonstrated that meso-FAP CAR-TEAM cells kill pancreatic cancer cells and CAFs in vitro, in vivo, and in patient-derived ex vivo models and have superior anti-tumor function compared to meso-CAR T cells alone. For the UG3 phase of this project, we will further optimize meso-FAP CAR-TEAM cells by determining the best mesothelin binder to use (SS1 vs. a novel binder developed by our lab), optimal route of injection (IV vs. IP) for targeting pancreatic tumors, and rationale drug combinations that address CAR T cell limitations in solid tumors. We will improve antigen density using an ADAM17 inhibitor (INCB7839) to prevent mesothelin cleavage from pancreatic cancer cells, optimize CAR T cell killing and persistence using ibrutinib to polarize meso-FAP CAR T cells to a Th1/Th17 phenotype, and further prevent suppression by the tumor microenvironment using a PD1 inhibitor (pembrolizumab). These drugs will be singly combined with meso-FAP CAR-TEAM cells to determine which best promotes efficacy in our preclinical models. Collectively, these results will inform the design of a phase I clinical trial for pancreatic cancer patients with advanced disease. During the UH3 phase, we will determine the safety and tolerability of meso-FAP CAR-TEAM cells. We have chosen pancreatic cancer as our first solid tumor target due to the dismal prognosis of the disease, the high percentage of patients with mesothelin-expressing tumors, and the known role of CAFs in promoting tumor growth. While our primary objective will be to determine safety, we will also monitor patient outcomes (progression and survival) while performing correlative studies to determine CAR T cell phenotype and function. We will also monitor the tumor and tumor microenvironment for mechanisms of response or resistance, such as changes in antigen expression and immunosuppressive cells. Overall, this project will develop a novel CAR-TEAM design to target a solid tumor and its microenvironment while optimizing the trial design through rigorous preclinical testing. If successful, the meso-FAP CAR T cell product could be directly applied to other mesothelin-expressing solid tumors and the knowledge gained from the UG3 phase will inform on the critical aspects of CAR T cell function to optimize prior to initiating a clinical trial.

NIH Research Projects · FY 2025 · 2025-09

Project Summary Pulmonary hypertension (PH) due to lung disease (Group 3) is one of the most prevalent and most lethal classifi- cations of PH. Two critical issues impede the effective clinical care of this disease: 1) prolonged time to diagnosis, due in part to the challenge of identifying the symptoms of the disease separate from those of the primary lung disease; and 2) complications in treatment due to the presence of multiple etiologies and the corresponding absence of a one-size-fits-all solution. Bronchoscopy is a commonly-employed procedure in the diagnosis, moni- toring, and treatment of the lung diseases of this group, and with the right tools would offer the perfect opportunity to simultaneously investigate the presence and development of PH as well. Our long-term goal is to provide a clinically viable endoscopic imaging platform for the early detection and enhanced personalized treatment of PH in patients undergoing bronchoscopy. This platform, Mechanomicroscopy, has the potential to assess pulmonary arterial pressure and remodeling quickly and with minimal clinical friction during the bronchoscopic procedure, resulting in 1) a significantly shortened path to diagnosis that doesn’t rely on the development of symptoms; and 2) direct structural assessments of pulmonary arteries that are suspected to be relevant to determining effective treatment. The objective of this proposal is to develop and validate Mechanomicroscopy for the transbronchial assessment of pulmonary artery pressure and arterial smooth muscle thickening, and to perform these assess- ments across a diverse population of individuals with Group 3 PH. In Aim 1 we will implement technological developments for our system that will allow us to identify pulmonary arteries in real-time, which will aid in the clinical efficiency of our imaging. In Aim 2 we will evaluate our diagnostic imaging criteria in an animal model, comparing and correlating our measurements of pulmonary artery pressure and pulmonary arterial smooth mus- cle thickness with those of the gold standards of right heart catheterization and histology. We will conduct a human study in Aim 3 in which we will image patients who have already undergone right heart catheterization and who do not have severe PH, again comparing our pressure measurements with right heart catheterization for human translation and analyzing arterial smooth muscle thicknesses across the diverse population to inform future studies focused on personalizing care.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT Post-surgical pain (P-SP) affects up to 30% of U.S. patients and up to 50% for high-risk surgeries like thoracotomies and amputations, with global incidence ranging from 5% to 85%. P-SP imposes an economic burden of $560-$635 billion annually, including direct medical expenses and indirect costs like lost productivity and disability claims. Current treatment relies on generalized pain management protocols, leading to poor outcomes, higher chronic pain risk, and increased opioid use. A more personalized approach is urgently needed to improve treatment effectiveness by accounting for individual pain sensitivity and psychological factors. To elucidate the neural and psychological mechanisms underlying P-SP and to facilitate early identification of patients at risk for chronic pain, enabling more targeted interventions, the proposed secondary analysis project will develop an interpretable and predictive model for P-SP outcomes by 1) identifying the most predictive features of P-SP outcomes using multi-modal data (functional neuroimaging, psychological, behavioral, and medical history); 2) applying Shapley value analysis to the output of supervised machine learning models to interpret the contributions of each feature to the predictions, providing clinicians with patient-specific insights to improve pain management and support personalized treatment planning; 3) developing a time-series model to predict P-SP outcomes and 4) applying Shapley value analysis to identify key temporal patterns that influence these predictions. The expected outcomes of this highly innovative project include the identification of reliable biomarkers for P-SP, which will serve as a foundation for developing individualized pain management strategies. The findings have the potential to transform post-surgical care by improving the early detection of P-SP, optimizing treatment protocols, and reducing opioid use. Ultimately, this project aligns with the broader movement toward personalized medicine and precision healthcare, offering a comprehensive solution for enhancing patient outcomes and alleviating the long-term burden of chronic pain on the healthcare system. This directly supports one of the primary missions of the National Institute of Biomedical Imaging and Bioengineering (NIBIB). 2

NIH Research Projects · FY 2025 · 2025-09

Abstract Copy number variants (CNVs) involve deletions and duplications of genomic segments spanning more than 50 basepairs and represent one of the most penetrant sources of pathogenic variants in neuropsychiatric disorders, with myriad impacts on many other human phenotypes as well. However, the relative impact of CNVs at the resolution of individual genes, exons, or functional categories, and especially across diverse global populations, has never been systematically assessed in neuropsychiatric disorders at scale. This omission can be attributed to the technical barriers in CNV discovery as well as the lack of large-scale, diverse neuropsychiatric cohorts. Traditional cytogenetic methods for CNV detection, such as chromosomal microarrays (CMA), are relatively low- resolution, and have largely precluded gene and exon resolution analyses. Recent advances in sequencing with whole exome (ES) and whole genome sequencing (GS) have dramatically improved our resolution, including the discovery of exon and sub-exon level CNVs. However, neither GS nor ES are perfect. While GS can interrogate the whole spectrum of CNVs across frequency and size, it is expensive. ES on the other hand, though affordable, can only query the rare coding portion of the genome for CNVs. Promisingly, the blended genome exome (BGE) sequencing approach has recently undergone heavy development and rapid adoption in a number of large-scale, diverse neuropsychiatric sequencing efforts, including in the Populations Underrepresented in Mental Illness Association Studies (PUMAS) project, NeuroDev, and Akili studies. BGE is composed of a high coverage ES (~30x) with a low coverage GS backbone (~2-3x), at a cost comparable to traditional exome sequencing. With this blend, BGE has delivered on marrying the affordability of ES with the full range of variant detection of GS when used to detect single nucleotide variants (SNVs) across the entire genome. Leveraging our expertise in computational methods development for CNV detection and association across GS and ES, we believe that in addition to SNVs, BGE is the perfect platform to capture the full range of CNVs across the genome at: a significantly improved resolution compared to CMA and ES; a significantly lower reference-bias compared to CMA; and a dramatically lower cost compared to GS. To achieve this, we will extend our GATK-gCNV pipeline for rare CNV detection in conjunction with our ancestry-aware SV imputation pipeline for use with BGE data. Preliminary results have already shown great promise. We will apply this pipeline to the more than 110,000 available BGE samples across PUMAS, NeuroDev, and Akili to generate a large-scale, diverse CNV callset. These variants will be made publicly available and can immediately be leveraged to significantly advance our understanding of the genetic architecture of neuropsychiatric conditions, especially in context of diverse genetic ancestry groups.

NIH Research Projects · FY 2025 · 2025-09

Fibromyalgia (FM) is a chronic debilitating painful condition, affects millions of people in the U.S. alone, and is often refractory to the current treatment. The etiology of FM is multifactorial in which deranged gut homeostasis has been strongly implicated. Up to 70% of FM patients exhibit signs of deranged gut homeostasis, highlighting a key gastrointestinal component in the pathogenesis of FM. To date, the molecular link between deranged gut homeostasis and FM is poorly understood. The gut brush border enzyme intestinal alkaline phosphatase (IAP), encoded by Akp3 gene in mice and Alpi gene in humans, is a critical regulator of gut homeostasis. We and others have reported that knockout of Akp3 (Akp3-/-, IAP deficiency) in mice disrupts gut homeostasis with increased gut and systemic inflammation markers. Our preliminary data demonstrate that a) Akp3-/- mice recapitulated FM- like phenotypes including i) hypersensitivity to mechanical and thermal stimulation, ii) spontaneous pain behavior, and iii) FM-related comorbid behaviors including depression and anxiety; b) administration of exogenous recombinant IAP reversed IAP deficiency and improved FM-like phenotypes in Akp3-/- mice; c) macrophages and transient receptor potential vanilloid 1 (TrpV1) expression were increased in the dorsal root ganglion (DRG) of Akp3-/- mice; and d) IAP levels were reduced in stool samples of FM patients as compared to healthy subjects. These findings suggest that IAP could be a key molecule link between deranged gut homeostasis and the pathogenesis of FM. We propose that deficiency of Akp3-encoded IAP would lead to FM-like phenotypes through macrophage-mediated sensitization of TrpV1 positive (TrpV1+) nociceptors in the DRG. In Aim 1, we will test the hypothesis that IAP deficiency induces macrophage infiltration into the DRG and FM-like phenotypes. We will examine i) macrophage infiltration into the DRG using flow cytometry, immunostaining, and two-photon imaging with an innovative in vivo DRG window technique to correlate with FM-like phenotypes; ii) the effect of reversing IAP deficiency with exogenous recombinant IAP on macrophage infiltration into the DRG and FM-like phenotypes; and iii) the relationship between IAP deficiency-induced deranged gut homeostasis and DRG macrophage infiltration. In Aim 2, we will test the hypothesis that macrophage infiltration into the DRG plays a critical role in FM-like phenotypes. Genetic and pharmacological approaches will be used to i) deplete systemic vs. DRG macrophages; ii) inhibit monocyte/macrophage trafficking; and iii) suppress inflammatory macrophages. In Aim 3, we will test the hypothesis that TrpV1+ nociceptors in the DRG are sensitized via macrophage- nociceptor interactions in mice with FM-like phenotypes. We will i) inhibit TrpV1 genetically (TrpV1DTR) or pharmacologically and ii) examine a macrophage-mediated signaling in sensitization of TrpV1+ nociceptors using an innovative DRG calcium imaging platform, TrpV1::ChR2 mice, and patch clamp recording. This project will provide insights into a novel molecular mechanism of FM pathogenesis and help transform the current treatment limited to managing FM symptoms to mechanism-based therapy for FM patients with deranged gut homeostasis.

NIH Research Projects · FY 2025 · 2025-09

Over the past two decades, advances in prenatal diagnostics have improved clinical outcomes by transitioning from low-resolution karyotyping and chromosomal microarray (CMA) to more recent methods like exome sequencing (ES) and genome sequencing (GS). However, diagnostic testing requires invasive procedures, such as amniocentesis, which carry risks to both maternal and fetal health and contribute to high healthcare costs. Non-invasive prenatal testing (NIPT) has mitigated these risks by allowing the detection of chromosomal aneuploidies from maternal blood plasma using cell-free fetal DNA (cffDNA). Despite its widespread adoption—screening 25-50% of pregnancies in the US — NIPT remains limited to low-resolution testing, primarily detecting aneuploidies and changes in a few targeted loci. We propose to validate and optimize our recently published high-resolution non-invasive fetal sequencing (NIFS) technology that is capable of unbiased fetal exome screening at high coverage from a maternal blood draw alone. This innovation could transform maternal-fetal medicine (MFM) and significantly reduce healthcare costs by obviating the need for invasive procedures in fetal genetic testing. Our preliminary studies have demonstrated feasibility of NIFS, with 95.7% sensitivity and 94% precision in surveying fetal genomes across gestational ages relevant to prenatal testing. This project aims to implement and further validate NIFS for potential clinical application through three main objectives. First, we will conduct a comprehensive validation study of anomalous pregnancies (AIM 1) through analysis of 650 retrospective pregnancies with fetal structural anomalies (FSAs) and matched GS from invasive testing to validate pathogenic variant prediction. These analyses will benchmark the efficiency of NIFS in the detection of single-nucleotide variants (SNVs), indels, and copy number variants (CNVs). Second, we will focus on optimization (AIM 2) by developing robust analytic pipelines to discover, interpret, and validate variants at all gestational ages relevant to prenatal testing, conducting stress testing for scalability and estimating unbiased specificity and sensitivity for NIFS as a potential replacement for invasive genetic testing and next step in high-resolution screening for all pregnancies. Third, we will explore the performance of NIFS in non-anomalous pregnancies (AIM 3) by evaluating 500 pregnancies that underwent invasive testing and subsequent GS without an FSA or indication for testing, and 100 pregnancies with aneuploidies or CNVs detected by NIPT. We will estimate NIFS performance metrics and added diagnostic yield over NIPT from these data. This research unites interdisciplinary experts from technological innovation, computational genomics, clinical genetics, and MFM. Our team will rigorously assess NIFS as a non-invasive prenatal screening method that could transform pregnancy care by providing unprecedented access to genetic data and guiding treatment strategies for expectant women.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY: A diagnosis of Young Onset Dementia (YOD) is emotionally difficult and often delayed because these conditions are uncommon and many patients exhibit early symptoms atypical for dementia (e.g., personality changes, reduced motivation and challenging behaviors). The emotional distress of the diagnosis process is amplified after a diagnosis is made due to role changes, relational conflict, poor prognosis, and lack of impactful treatments. If untreated this early emotional distress becomes chronic and impairs patients’ and care-partners’ ability to work together to plan for the considerable adjustments required as the patient experiences loses to independence, function and roles in the prime of their lives. With a supplement to R21 NR01797902, we developed and established the feasibility of Resilient Together-YOD (RT-YOD), the first dyadic (patient and care-partner together) resiliency program for patients with YODs and their care-partners. RT-YOD draws on mechanisms from the dyadic program “Recovering Together,” an NIH funded, feasible, well-accepted program associated with sustained improvement in emotional distress among dyads after acute brain injury. We now propose to conduct a single blind, virtual RCT (NIH stage 2) of RT-YOD versus a Health Enhancement Program control (HEP; N=194 dyads), to establish the efficacy and durability (3 months) of RT-YOD on emotional distress (primary outcome) and quality of life. We will also examine whether improvements occur through putative mechanisms and are moderated by relevant clinical and demographic factors. Our team is uniquely positioned to conduct this study with MPIs (Vranceanu, psychologist; Dickerson, neurologist; Syme, early stage geropsychology investigator) with prior collaboration on R21NR017979-02S1, complementary expertise and established infrastructure to allow for timely recruitment of dyads.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT Up to one-third of older U.S. adults may not return to functional baseline six months after major surgery, with postoperative pain being one of the most significant contributors. At the same time, over a third of the adult population also suffer chronic sleep problems, which are even more prevalent in the 3.5 million adults who will require total knee arthroplasty (TKA) by 2030. Despite evidence-based treatment, perioperative sleep optimization represents a missed opportunity for improvement. One challenge is that the underlying mechanisms of how sleep impacts postoperative recovery remain a significant knowledge gap. Recent evidence suggests poor sleep health may impact pain via the nervous system and gut microbiome. Major surgery is a significant stressor, and habitual sleep disruption results in changes to metabolism, neuroinflammation, or the nervous system that increase the pro-nociceptive response to this stress. The gut microbiota influences host homeostasis for these processes, and insomnia can disrupt the gut-brain axis. Sleep health metrics of low sleep efficiency and poor daytime alertness were linked to increased blood homocysteine and IL-6 levels and decreases in the short-chain fatty acids-producing microbiota. At the same time, gut microbiome composition is altered in chronic pain conditions. Thus, the gut microbiome may mediate the influence of sleep on pain and serve as a target for adjuvant therapy for both pain and insomnia. My research program has built a unique translational perioperative sleep infrastructure for prospective studies and randomized efficacy trials. We focused on developing feasible and scalable at-home methods to estimate multi-dimensional sleep and circadian phenotypes before surgery using wearable actigraphy watches and other portable home devices. We have also collected blood to verify the circadian phase (internal time) using transcriptomic assays utilizing the circadian nature of transcriptomes. We propose using these preoperative sleep/rhythm phenotype measures to optimize surgical recovery. This is my area of expertise; I am an anesthesiologist with formal training in sleep/circadian biology, computational neuroscience, behavioral and psychosocial clinical trials, and translational studies in post-surgical pain and cognition outcomes. This research plan will aim to understand the mechanisms of how preoperative sleep/circadian disturbances may impact recovery outcomes through systemic changes in the gut microbiome and related metabolic factors. To address this, we propose 1) a longitudinal study (pre- to 1-year post-surgery) with rich phenotyping to understand the sleep-gut-pain relationships in TKA patients and 2) a randomized mechanistic trial with a well-known non-pharmacological intervention (CBT-I) to test whether intentionally altering real-world sleep conditions improves microbiome features. This may help us better risk stratify surgical patients and gain mechanistic inferences to novel mechanisms that inform therapeutic targets.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY / ABSTRACT Evidence is slowly building for a more nuanced and complex architecture within human and macaque lateral geniculate nucleus (LGN) than the classic view of three cell classes separated into six laminae. Exhaustive investigations in primate retina have revealed a highly diverse set of retinal ganglion cell (RGC) types that project to the LGN; while the majority of projections by fiber count are from the primary classes of RGCs (midget and parasol) that target the magno- (M) and parvocellular (P) layers in LGN, many of the classes with sparser fiber count (bistratified, thorny, sparse, monostratified, etc.) project to the koniocellular (K) layers, or have undescribed targets in the LGN. Recent single cell RNA sequencing (scRNA-seq) results on neuronal nuclei microdissected from differ- ent layers of LGN suggest a broader diversity than currently understood by classical anatomical and funct- ional results, with eight cell types identified in macaque: four GABAergic cells (presumably interneurons), M, P, and two kinds of K cells. Our analysis of data from the literature, presented here, suggests that additional sub-classes can be identified. But the spatial distribution of those cell types, specifically the multiple GABAergic and K classes, and the novel sub-classes we have found, is as-yet unknown. We will fill that gap by applying the recently available tools of spatial transcriptomics to analyze the layout of geneticaly-defined cell classes across the LGN, creating data sets from human autopsy tissue. These data will be generated in collaboration with the UCLA Technology Center for Genomics and Bioin- formatics on their NanoString CosMx SMI instrument. Human tissue will be obtained in collaboration with the MGH Pathology Department through an unrelated program that requires rapid autopsy following death. These data will allow us to test hypotheses about the spatial distribution of the factors that determine the cellular subtypes above, such the NOTCH developmental pathway, NDUF, COX, and ATP metabolic pathways, STX1A signaling pathway, and others. We will test whether different LGN laminae within the M, P, and K pathways are genetically differentiated, along with variation along eccentricity through anterior vs posterior sections, or along projection columns. The results from this initial project will be used as preliminary data in support of obtaining funding for a larger, more extensive study that will look for variation within and across individuals, asking questions about cell type distribution across the visual field, within and across laminae, then between sexes, through age, and finally about changes due to retinal or other blinding diseases.