Brigham And Women'S Hospital

NIH Research Projects · FY 2026 · 2024-09

Project Summary Migraine, a chronic intermittent headache disorder, ranks in the top five causes for years lived with disability. Approximately 15% of the US population experiences migraine, with women afflicted approximately twice as often as men. Although pharmacologic medications are often used as first-line treatments for migraine, these treatments may have difficult side effects and may increase the risk for migraine chronification. As a result, individuals with migraine often turn to other treatment modalities, including the use of natural products and dietary supplements, for long-term migraine management. Preliminary evidence from the COcoa Supplement and Multivitamin Outcomes Study (COSMOS) suggests a cocoa extract supplement may have beneficial effects on migraine. COSMOS was a randomized, double-blind, placebo-controlled trial testing a cocoa extract supplement containing 500 mg cocoa flavanols/d (including 80 mg (-)-epicatechin, plus 15 mg caffeine and 50 mg theobromine) for the prevention of cancer and cardiovascular disease in 21,442 older adults. Every six months, participants were asked about the occurrence of adverse events, including migraine. Individuals randomized to the cocoa extract supplement were significantly less likely to report migraine (hazard ratio (HR)=0.85; 95% confidence interval (CI): 0.78, 0.93) than those assigned to placebo. However, several important gaps in knowledge remain. First, COSMOS enrolled only older adults but the prevalence and disability burden of migraine is highest at younger ages (i.e. 15-49 years) and it is unknown if cocoa extract is effective in a younger population. Second, COSMOS did not collect detailed information on changes in migraine frequency, as recommended by current guidelines for trials of migraine treatments. Finally, an animal study suggested a potential dose-response relationship between cocoa consumption and migraine pathophysiology, highlighting the importance of considering higher amounts of cocoa extract and bioactive content in future human studies. Our long-term goal is to conduct a fully powered trial evaluating the effectiveness of a cocoa extract supplement on reducing the frequency of migraine attacks. As a first step, we propose to conduct a three-arm pilot study. We will recruit and randomize (1:1:1 allocation ratio) 114 adults with episodic migraine to receive one of the following treatments for 12 weeks: 1) 1000 mg of cocoa extract (including 160 mg/d (-)-epicatechin, 100 mg theobromine, and 30 mg caffeine); 2) 500 mg of cocoa extract (including 80 mg/d (-)-epicatechin, 50 mg theobromine, and 15 mg caffeine); or 3) placebo. Our proposed pilot study will allow us to address the following aims: 1) to assess the feasibility of recruitment, retention, and adherence; 2) to determine the acceptability of higher doses of cocoa extract supplement to this patient population; 3) to optimize data collection and data management and establish the infrastructure needed for a large-scale trial.

NIH Research Projects · FY 2025 · 2024-09

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. Leveraging the past success of Harvard Medical School’s (HMS) affiliated four major teaching hospitals and basic science departments, the Professional Development Core (PDC) will focus on preparing trainees for a smooth entry into the program and supporting them with a series of resources and individualized opportunities to optimize research and career development experiences within the program. This Core will produce a tailored combination of formal programs, seminars, and networking events, and personal interactions with faculty and other trainees including opportunities for introduction to and acquisition of data analytic and other technical skills integrated with an online informational and learning hub. Trainees will be exposed to and explore complementary sciences and technologies through intentional interactions with peers and mentors in other research areas, creating multiple cross-discipline opportunities. These interactions are designed to inspire innovation and promote entrepreneurship. The PDC will support the ongoing development and training of trainees and mentors through the following Aims. Aim 1: Coordinate the selection of trainees, composition of Individual Oversight Committees (IOCs) and Individual Training Plans (ITPs). The PDC will adopt a structured approach to assist trainees in project and mentor selection, and in formation of the IOC. Emphasis will be placed on trainee educational goals ensuring an individualized training experience, and an ITP will be created with input from mentors, trainees, and technical experts. Formal evaluative tools will be used to monitor trainee experiences, trainee progress and mentor effectiveness which will inform the evolution of the overall program. Aim 2: Develop and present specific course curricula, interactive events, and materials to enhance trainee education. In collaboration with the Network Core, the PDC will develop new didactic experiences in areas important for the immediate and long-term success and competitiveness of academic researchers with an introductory KUH minicourse, monthly seminar series and annual retreat. These will benefit from input from the Industry/Entrepreneurism Council and Patient Input Council. The Core will also leverage existing ecosystem resources summarized on the HKUHTI web Hub to ensure training in scientific writing, mentoring and team science. Aim 3: Provide technical electives that facilitate incorporation of state-of-the-art technologies into trainee career development. The PDC will offer a variety of curated opportunities to facilitate trainee competence in data analytics and large databases, as well as exposure to novel technology platforms to enhance training in cross-disciplinary research. These interactions will enhance the trainee’s professional network and create opportunities for interdisciplinary connections among Kidney, Urology and Hematology disciplines. The PDC will work closely with other Cores to develop multi-disciplinary trainees at various stages of career development with a deep passion for innovation and team-based KUH research.

NIH Research Projects · FY 2024 · 2024-09

Project Summary/Abstract Early detection and sensitive tracking of cognitive changes related to Alzheimer’s disease (AD) pathology (amyloid-Aβ and tau), particularly those that are scaleable to the large at-risk population of older adults, are urgently needed. Promising digital tools to measure cognitive changes must be validated in demographically diverse groups to ensure that research advancements are generalizable to those most at-risk for cognitive decline. Our recent work suggests that diminished learning over repeated evaluations (i.e., testing memory for the same content for 12min/day for 7 days) reveals subtle Aβ-related memory failures that are otherwise undetectable using single timepoint measures. More specifically, 29 Aβ+ clinically unimpaired (CU) older adults failed to improve at the same rate as their 97 Aβ- CU peers when asked to learn and recall identical content daily with this group difference emerging after ~3 days. We now seek to determine whether this Personal Learning Curve (PLC) paradigm can be used to detect early memory failures related to AD biomarkers in a larger and more diverse and representative sample and whether PLCs can sensitively track longitudinal cognitive decline when repeated bi-annually. We have the unique opportunity to recruit an extremely well-characterized and representative at-risk sample (i.e., 32% from URG, family history of cognitive decline, hypertension, geographic diversity, English and Spanish speaking) with AD biomarkers (global Aβ, temporal tau quantified via Positron Emission Tomography-PET) and annual cognitive testing. Recruited participants (n=300) will have completed the active phase of U.S. POINTER, a two-year clinical trial of two different multi-domain lifestyle interventions to protect brain health. We will explore whether a PLC, collected 12min/day for 7 days on personal devices, is a valid and sensitive marker of memory decline in this representative population and whether repeating a PLC every 6 months for 12 months will reveal AD-relevant longitudinal cognitive decline. In exploratory analyses, we will examine PLCs in relation to cardiovascular risk factors (e.g., systolic blood pressure, white matter hyperintensities via Magnetic Resonance (MR) Imaging) and non-specific markers of neurodegeneration (e.g., entorhinal tau, hippocampal atrophy). If successful, this proposal will provide a rapidly obtainable, repeatable, high-resolution snapshot of clinically relevant memory failures to facilitate early detection of cognitive decline as well as provide a novel method to assess treatment response more rapidly. Doing so in a representative population will ensure that the most promising novel and accessible digital cognitive tools are generalizable to the broader at-risk population.

NIH Research Projects · FY 2025 · 2024-09

The methods used to measure NAD+ and NADH in human samples include the enzymatic cycling assay, high pressure liquid chromatography and liquid chromatography tandem mass spectrometry (LC-MS/MS). The LC- MS/MS method offers superior sensitivity, precision, accuracy compared to the other methods. However, all current methods suffer from one or more limitations: 1) some sample processing methods use strong acids (e.g., trichloroacetic acid or perchloric acid) which degrade NADH and NADPH; 2) separate methods are often used to measure oxidized and reduced pyridine nucleotides which poses a barrier to measuring the full spectrum of NAD+-related metabolites; 3) scarcity of stable isotope-labeled internal standards for NAD+-related metabolites that are imperative for precise quantification of analytes; 4) Commutable reference materials used to calibrate assays and provide accuracy, metrological traceability, and comparability of measurements by different laboratories are not available, nor are there mechanisms to harmonize assays across laboratories. We propose two integrated aims to fill these methodological gaps. In Aim 1 we will (a) optimize and validate standardized sample collection and storage protocols that will preserve the integrity of the pools of NAD+, NADH, NADP, NADPH and related metabolites; (b) finalize the validation of our advanced LC/MS-MS method (NADome Quant) that simultaneously measures this set of metabolites in human tissues; and (c) in collaboration with Dr. Vesper's laboratory at the CDC, generate higher order commutable reference standards and establish processes for harmonizing assays across laboratories. In Aim 2, we will conduct a randomized, double-blind, placebo-controlled trial (n=90) to determine whether oral administration of β nicotinamide mononucleotide (NMN), an immediate NAD+ precursor, is more efficacious effective than placebo in raising myocardial NAD+ levels and reducing myocardial injury in adults undergoing elective coronary artery bypass graft (CABG) surgery. Patients will be randomized 1:1:1 to placebo or to one of two dosing regimens of NMN (administered for seven vs. two days prior to surgery) to test whether NMN administration increases myocardial concentrations of NAD+ and its metabolites and attenuates myocardial and extracardiac (e.g., kidney) injury. CABG surgery offers a unique opportunity to harvest a variety of human tissue that would otherwise be discarded, including myocardium, aortic endothelium, subcutaneous fat, and skin. This research proposal will establish robust sample collection and storage protocols that preserve NAD+ as well as NADH and NADP(H); more advanced LC/MS assays for measuring NAD+ and related metabolites; and commutable reference materials and procedures for harmonizing assays across laboratories through the auspices of the CDC. The proof-of-concept trial will provide key foundational data to guide the design of larger efficacy trials of NAD+ augmentation strategies for preventing myocardial and extracardiac injury during CABG surgery, an important unmet clinical need.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Heart failure affects 6 million Americans, 1.8% of the US population. Despite substantial progress, heart failure mortality remains ~50% at 5 years. Mechanisms of cardiac dysfunction in heart failure remain poorly understood. In this project, we will characterize two cardiac diseases involving mechanisms of infiltration and toxicity: light- chain amyloid cardiomyopathy (AL-CM), and cancer-therapy-related cardiac dysfunction (CTRCD). AL-CM results from plasma cell clones producing abnormal immunoglobulin light chains, which aggregate as amyloid fibrils in the heart. Median untreated survival is <6 months. CTRCD appears in 7–65% of patients undergoing anthracycline chemotherapy. One-year mortality is >50% in symptomatic patients. There is an unmet need for a better understanding of mechanisms of cardiac dysfunction, including the reasons that abnormal light chains or toxic chemotherapy cause cardiac dysfunction in some individuals and not in others. Our scientific premises are that (1) cardiac dysfunction is accurately detected and quantified by imaging, and (2) cardiac dysfunction alters circulating levels of proteins involved in pathogenic mechanisms. Therefore, our overall approach is to join cardiac imaging and plasma proteomics to identify circulating proteins from relevant mechanistic pathways and to characterize their relationships with imaging metrics of cardiac dysfunction. To achieve the aims of this research project, my advisory team and training plan will cover cardiac imaging, proteomics, AL-CM, CTRCD, and data science. Our first aim is to identify circulating proteins involved in mechanisms of AL-CM by comparison to non-amyloid participants and by correlation with imaging metrics of cardiac dysfunction. In the K99 phase, we will analyze a panel of plasma proteins and compare their levels in participants with AL-CM vs. multiple myeloma (abnormal immunoglobulins without amyloidosis). Then, we will measure correlations between protein levels and cardiac amyloid burden by 18F-florbetapir PET, extracellular expansion by extracellular volume (ECV) on MRI, and contractile dysfunction by global longitudinal strain on echocardiography. Our second aim is to identify novel markers of CTRCD by group comparisons of plasma proteins levels and by correlations of protein levels with changes in imaging metrics of cardiac dysfunction. In the R00 phase, I will analyze a panel of plasma proteins and compare their levels in participants undergoing anthracycline chemotherapy with CTRCD, without CTRCD, and in matched participants with heart failure. Then, I will measure correlations between changes in protein levels and changes in ejection fraction or in ECV by MRI over 12 months. Lastly, I will compare baseline protein levels in patients with vs. without subsequent CTRCD. The successful completion of this project will support my goal of becoming an independent clinician-researcher on heart failure, deepen our mechanistic understanding of AL-CM and CTR-CM, improve risk stratification and prevention strategies, and identify novel drug targets to improve the outcomes of patients with heart failure.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT Immune checkpoint inhibitors (ICIs) have drastically improved cancer survival over the past decade, but this survival comes at the cost of a new class of immune-related adverse events (irAEs) characterized by inflammatory and auto-immune pathologies that occur and persist long after ICI discontinuation. These irAEs can have major impacts on long-term quality-of-life, but our ability to appropriately address them is limited by an insufficient understanding of irAE rates and severity profiles. Automated methods to identify and monitor irAEs could improve clinical care, biomedical research, and pharmacovigilance, however, irAEs are often only documented in clinical text and cannot currently be automatically extracted from the EHR at scale. The overarching objective of this proposal is to create applied informatics technologies for cancer surveillance research and survivorship care in patients treated with ICIs. Our central innovation is the development and clinical validation of natural language processing methods, particularly neural language models, that can handle the complexities of the EHR for irAE extraction using unstructured and structured data streams. In Specific Aim 1, we conduct a clinical trial of informatics-assisted irAE detection from the EHR, measuring feasibility and effectiveness in improving registration onto Alliance A151804, an NCI cooperative group irAE biorepository. This will be the first trial of informatics-based adverse event detection for cancer care and a major step toward clinical translation. In Specific Aim 2, we develop new methods to extract irAEs according to their severity grade for detailed and standardized computational phenotyping, and perform external validations. In Specific Aim 3, we optimize generalist large language models for irAE information extraction without task- specific fine-tuning, including innovative methods to tailor models’ diagnostic reasoning to each patient. This work is highly significant for developing, applying, and validating informatics methods that take full advantage of the EHR to support the long-term goal of improving quality-of-life and survival in patients treated with ICIs. This clinical translational work will be carried out by an expert team of cancer clinicians, clinical trialists, informaticians, and computer scientists.

NIH Research Projects · FY 2026 · 2024-09

Enter the text here that is the new abstract information for your application. The goal of the Harvard Kidney, Urology and Hematology Training Institute’s (HKUHTI) Training Program is to educate the next generation of leaders of KUH research by leveraging the intellectual, collaborative, and physical resources of our rich Boston ecosystem. Five partnering institutions request support for a TL1 Training Core to fund 10 pre- and 24 postdoctoral trainees in the steady state of the grant (after some adjustments in the first year taking into account the end of existing T32 grants). Trainees will benefit from HKUHTI-specific, as well as collaborative training curricular offerings that will be tailored to the trainee’s individual scientific and career goals. The HKUHTI will employ a rigorous selection process that will result in appointment of a scientifically-varied group of trainees from across the country to a program that provides rigorous training with a world-class roster of mentors in KUH fields. Trainees will benefit from a primary mentor and an individual oversight committee (IOC) throughout their tenure in the program. The HKUHTI will expose trainees to a range of innovative research opportunities, including those offered by established mentors, and will leverage a clinical and translational research accelerator, the Harvard Catalyst. The TL1 will address many identified challenges to training the next generation of KUH research leaders and will build on a strong history of success in developing and retaining leaders in KUH biomedical sciences. The requested positions will bring together K, U, and H across pediatric and adult divisions. The TL1 is closely linked to the U2C, incorporating rich Professional Development and Network Cores, and the supported trainees will benefit from a robust Administrative Core with a digital Hub, oversight committees, and evaluation infrastructure. Formalized structures are established for recruitment, selection, and retention, as well as mentor and trainee selection and evaluation. The TL1 will focus on offering trainees the knowledge needed to perform state-of-the-art science and prepare for a productive and rewarding career in KUH fields. Additionally, the TL1 is structured to provide oversight to trainees and ensure that the trainee, mentor(s), and IOC co-develop an Individualized Training Plan in a cooperative manner, such that the needs, emerging research interests, and future career of the trainee are always at the center of their training experience. The structure of the TL1 program was intentionally designed to address seven major challenges identified by surveying the trainees (see Training Plan). In addition, the HKUHTI will benefit from validated evaluation tools to collect data and address emerging trends and scientific challenges to ensure that trainees, mentors, and the entire HKUHTI community achieve measurable personal, professional, and scientific growth.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY / ABSTRACT Heart failure with preserved ejection fraction (HFpEF) affects 2.5 million Americans and is associated with high morbidity and mortality. Few effective treatments are available. This proposal will investigate an important cause of HFpEF: myocardial interstitial fibrosis (MIF). Hypertension, diabetes, and systemic inflammation lead to MIF by activating fibroblasts, which secrete regulatory proteins that deposit, process, and cross link collagen fibers in the myocardial extracellular matrix. Treatment of MIF is a potential strategy to prevent or treat HFpEF, but will require scalable diagnostic tests and proven anti-fibrotic therapies. Dr. Cunningham, a heart failure cardiologist and early career investigator at Brigham and Women’s Hospital in Boston, proposes to study MIF using cardiac MRI, large-scale proteomics, and clinical trial methods. Preliminary data show that inhibition of neprilysin (an enzyme which degrades natriuretic peptides) by the heart failure drug sacubitril/valsartan reduces plasma levels of several extracellular matrix regulatory proteins. These results suggest that neprilysin inhibition may attenuate MIF. However, it remains unknown 1) whether plasma levels of extracellular matrix regulatory proteins are associated with MIF, 2) which extracellular matrix regulatory proteins are modified by neprilysin inhibition, and 3) whether neprilysin inhibition reduces or slows progression of MIF. In this proposal, Dr. Cunningham hypothesizes that plasma levels of extracellular matrix regulatory proteins identify MIF and that neprilysin inhibition reduces MIF through its effect on these proteins. In Aim 1, he will evaluate whether plasma levels of pre-specified extracellular matrix regulatory proteins are associated with MIF measured by cardiac MRI in community participants from the UK Biobank. In Aim 2, he will investigate the effect of neprilysin inhibition on plasma levels of these proteins in a previously conducted HFpEF randomized trial. In Aim 3, he will conduct a pilot randomized trial directly assessing the effect of neprilysin inhibition on the most reproducible marker of MIF, extracellular volume measured by contrast cardiac MRI. If successful, these studies will form the foundation for future R01 applications identifying patients with MIF using proteomics and cardiac imaging and evaluating the benefit of neprilysin inhibition or other anti-fibrotic therapies in these patients. These specific aims are part of a comprehensive career development program designed to provide Dr. Cunningham with the skills to become an independent R01-funded physician-scientist. Dr. Scott Solomon, an expert in heart failure clinical trials at Brigham and Women’s Hospital, and Dr. Patrick Ellinor, an expert in cardiovascular epidemiology at the Broad Institute, will mentor Dr. Cunningham. Dr. Cunningham will develop expertise in the biostatistical analysis of large-scale physiological data, the interpretation of cardiac MRI, and the execution of heart failure clinical trials. Dr. Cunningham’s long-term goal is to lead clinical trials that leverage physiological data to precisely target therapies to individual heart failure patients.

NIH Research Projects · FY 2025 · 2024-09

Summary/Abstract Asthma is a common chronic respiratory disease and is considered to be an umbrella diagnosis for several sub-diseases with distinct biological pathways and clinical presentations. Moreover, asthma has a strong genetic predisposition and is associated with early-life factors and environmental exposures. Previous studies identified several associations between asthma and omics factors, but the exact mechanisms underlying most findings remain unclear. Multi-omics approaches were proposed as a tool to dissect asthma endotypes and heterogeneity. In this application, we aim to utilize multi-omics data to disentangle asthma heterogeneity, identify endotypes, and investigate biological mechanisms underlying established risk factors. The analyses will take advantage of three cohorts of (asthmatic) children with high-quality multi-omics data and extensive phenotype information. In Aim 1, we will analyze the interplay between genetic, epigenetic, and metabolomic factors in the development of asthma. In Aim 2, we will identify asthma endotypes based on genetically predicted multi- omics levels, directly inferring the genetic basis of these distinct mechanisms. Furthermore, we will investigate omics signatures of parental asthma history at birth and during childhood (Aim 3). We will characterize the clinical and molecular features associated with significant results throughout the aims. The aims require innovative methodological advances derived as part of the project to unfold the full potential of the data resources and utilize the unique advantages of the family-based designs. Overall, achieving the research objectives demands skills covering respiratory biology, statistics, multi-omics expertise, and data science/causal inference. Julian Hecker, Ph.D., MS, is a statistical geneticist whose long-term career goal is to establish himself as an independent research scientist in the study of asthma and respiratory diseases. The detailed career development plan will support Dr. Hecker to achieve the following training goals and close knowledge gaps: 1.) gain expertise and experience in integrative omics analyses and the processing of multi-omics data, 2.) deepen understanding of asthma pathophysiology, and 3.) improve apprehension of causal inference techniques in epidemiological analyses. Dr. Hecker’s strong theoretical background, in combination with his experience with family-based approaches and analyses, will position him to accomplish these training goals and the aims of this proposal, as well as prepare him for the transition to an independent research role. He will take advantage of the unique research environment in the Channing Division of Network Medicine and Harvard Medical School. The support from his experienced mentoring team and the advisory committee will ensure the proposed project's success.

NIH Research Projects · FY 2025 · 2024-09

Project Summary Anti-retroviral therapy (ART) has protected many HIV patients against the development of AIDS. HIV infection has become a treatable disease. Increased risk for atherosclerotic vascular diseases in long-term HIV survivors or people living with HIV (PLWH) with no detectable virus load, however, is projected to become a global health burden. Despite its clinical impact, underlying mechanisms for accelerated atherogenesis in PLWH remain obscure. Macrophages promote the formation of high-risk plaques prone to vascular events (e.g., macrophage-rich, large necrotic core, think fibrous cap). Macrophages are heterogeneous and the imbalance of their subpopulations may accelerate atherogenesis. Evidence suggests that extracellular vesicles (EVs) that contain the HIV-associated protein Nef (Nef EVs) promote chronic inflammation. We will examine the effects of Nef EVs in the heterogeneity and functions of macrophages and the formation of atherogenesis. Our study represents a dynamic interplay of biology and data science to identify novel mechanisms and therapeutic targets. We will use a systems approach to test the central hypothesis that Nef EVs modulate macrophage heterogeneity, shifting the balance toward an atherogenic or less atheroprotective phenotype. Our preliminary data have led us to the specific biological hypothesis that Nef EVs impair efferocytosis and contribute to the formation of high-risk atherosclerotic plaques that we will test in three Specific Aims. In Specific Aim 1, we will conduct a systems-based macrophage profiling, involving unbiased multi-omics, data integration, and network analysis to identify novel mechanisms for macrophage activation by Nef EVs. Pilot multi-omics data suggested that Nef EVs suppresses efferocytosis. In Specific Aim 2, we will validate omics data in vitro and in vivo and address underlying mechanisms for impaired macrophage efferocytosis by Nef EVs.

NIH Research Projects · FY 2025 · 2024-09

ABSTRACT Women with severe aortic stenosis (AS), a condition characterized by the narrowing of the aortic valve opening, often experience delayed diagnosis, undertreatment, and higher mortality rates compared to men, indicating both delayed care-seeking and a lack of appropriate diagnostics and monitoring for female patients. Yet, the influence of anatomical and functional differences in the female population on AS presentation, management, and outcomes remains poorly understood. Furthermore, despite the prevalence of symptoms, women with severe AS receive less aortic valve replacement (AVR) treatment and have higher excess mortality rates over a five-year period compared to men. Our proposed project integrates innovative medical image processing and computational modeling methods, such as statistical shape analysis (SSA), convolutional neural networks (CNN), and inverse finite element analysis (FEA), to gain sex-specific insights into cardiac remodeling and dysfunction, with a specific focus on severe AS in women. By focusing on cardiac remodeling, a consequence of prolonged aortic valve disease, our goal is to enhance AS treatment for women by considering sex-specific differences in ventricular responses to AVR-induced afterload. To achieve this, we will develop a personalized, mathematical approach that leverages sex-differentiating anatomical and functional characteristics of the left ventricle (LV), ultimately aiming to improve survival outcomes. Additionally, we will compare the predictive value of these sex- differentiating measures to traditional indices, enhancing our understanding of their effectiveness in guiding clinical management. We hypothesize that advanced anatomical metrics (e.g., shape scores) and material characteristics (e.g., cardiac stiffness) are superior predictors of post-intervention cardiac events and dysfunction compared to traditionally collected clinical measures. Our research consists of two main aims. Aim 1 involves developing a fully automated, neural network pipeline to segment clinical images, creating an advanced SSA model to extract hidden geometrical features, and establishing a correlation between shape scores and post- intervention clinical events. This analysis will assess the predictive power of sex-specific measures compared to the male population, cases where sex is not considered in the model training, and universal clinical indices. Aim 2 focuses on developing a computational tool to estimate patient-specific stiffness of the inhomogeneous LV tissue, with an examination of its potential value in predicting diastolic dysfunction and AVR outcomes. Our research serves as a steppingstone to guide clinicians in preprocedural patient selection, optimize surgical timing, and improve survival outcomes. By developing a sex-specific risk stratification tool and a mechanistic framework for effective prognosis, we aim to provide valuable means to enhance treatment and mitigate devastating events associated with severe AS, particularly for female patients.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Glioblastoma (GBM), one of the most fatal of cancers, remains impervious to treatment with the current standard of care (SOC) therapies. Even immunotherapies like immune checkpoint inhibitors (ICI) have failed for GBM despite their success with other cancers. One critical reason for this failure is that the GBM tumor microenvironment (TME) is highly immunosuppressive. To change the GBM microenvironment, we and others have been utilizing direct intra-tumoral administration of gene therapies that express powerful immune-activators, both preclinically and clinically. In fact, we have concluded a phase 1 clinical trial of intratumoral administration of a regulatable interleukin 12 (IL12) immunogene therapy in subjects with recurrent GBM (rGBM)1 (NCT02026271). In this trial, post-injection tumors showed increased inflammatory infiltrates including increased numbers of CD8+ T cells and elevated levels of tumor IFNγ. However, this also was coupled with increases in tumor PD-L1 and PD-1 positive cells, characteristic of chronic exhaustion and immune escape. Therefore, the trial suggested that addition of anti-PD-1 immune checkpoint signaling should be beneficial to IL12 immunogene therapy efficacy. Based on this, we finished accrual to two multi-institutional clinical trials of intratumoral IL12 immunogene therapy (to increase infiltration of IFNγ producing, CD8+ cytotoxic T cells) coupled with neoadjuvant immune checkpoint inhibition (ICI) (NCT03636477 and NCT04006119). These trials have shown that the combination of these two immunotherapies is well tolerated in humans, but unexpectedly there was no improvement in efficacy by adding ICI. Based on this clinical result, we propose to explore two questions using preclinical models of GBM: 1- Does the timing of ICI administration with respect to IL12 immunostimulation change the therapeutic response? 2- Does GBM immune-evasion from IL12 depend on more than single PD-1 signaling? To provide answers to these two questions, we plan to pursue the following aims: Aim 1- Compare efficacy, tolerability, and immune effects of neo-adjuvant vs. adjuvant anti-PD1 therapy in mouse models of GBM treated with intratumoral IL12 immunogene therapy, and Aim 2- Test the efficacy, tolerability, and immune effects of silencing the immunoevasive noncoding RNA, INCR1, when combined with intratumoral IL12 immunogene therapy. The impact from these studies will be to significantly inform whether IL12 (or other cytokines) immunogene therapy can be improved by changing the timing of ICI administration or by targeting a different pathway that regulates multiple immunoevasive signals, leading us to the next design of clinical trials using cytokine immunogene therapy.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Low-resource settings have a high burden of Neisseria gonorrhoeae infection, but insufficient laboratory infrastructure for diagnostic tests and antimicrobial susceptibility determination, leading to syndromic management, which misses a high proportion of cases and results in the overuse of antibiotics. Together, those factors contribute to driving the emergence of antimicrobial resistance (AMR), a global public health threat. Two novel strategies can combine to address those challenges. First, the use of CRISPR-based technology permits the development of low-cost, point-of-care (POC) assays, which can be used in resource-limited settings. Second, determination of molecular resistance markers can facilitate resistance-guided therapy, thereby reducing the selective pressure towards AMR. The study aims are: 1) adapt and validate existing CRISPR-based assays for POC detection of N. gonorrhoeae and ciprofloxacin susceptibility, 2) develop a multiplexed CRISPR-based assay for N. gonorrhoeae cefixime susceptibility, and 3) evaluate the feasibility and acceptability of the POC N. gonorrhoeae and AMR assays among two health centers in Botswana. The study is based upon our preliminary work in the development of Cas13a-based diagnostics for N. gonorrhoeae and ciprofloxacin susceptibility, and will leverage clinical specimens collected as a part of an ongoing trial at the Massachusetts General Hospital Sexual Health Clinic, as well as through existing collaborations in Botswana. This Mentored Patient-Oriented Research Career Development Award (K23) supports the career development of Dr. Lao-Tzu Allan-Blitz, an instructor of medicine with experience in the epidemiology of and novel diagnostic assays targeting AMR sexually transmitted infections (STI). The K23 will support Dr. Allan-Blitz to develop expertise in 1) assay development for POC detection, and 2) assay implementation and evaluation. To achieve the proposed research and training aims, Dr. Allan-Blitz has assembled a team of mentors who are world experts. Dr. Jacob Lemieux, who is an expert in metagenomics, DNA sequencing, and CRISPR-based assay development, will serve as the primary mentor. In addition, his Co-Mentorship team includes Dr. Pardis Sabeti, a world expert in computational genetics, microbial genomics, diagnostic technologies, and disease surveillance in Africa, and Dr. Shahin Lockman, a world leader in implementation science research with longstanding collaborations in Botswana. The proposed research and training aims will leverage strong, longstanding collaborations and the robust research infrastructure between Brigham and Women's Hospital, Massachusetts General Hospital, the Broad Institute, and the Botswana Harvard AIDS Institute Partnership. Finally, the K23 award will advance Dr. Allan-Blitz's career goal of becoming an expert in assay development for AMR STI and assay implementation, as well as facilitate his transition to research independence.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY This K01 career development award will position the candidate to become an independent translational re- searcher studying mind-body movement therapies (MBMT) for chronic pain conditions linked to chronic inflam- mation, beginning with persistent post-surgical pain (PPSP) as a model. BACKGROUND. Incidence of PPSP varies according to surgical type, with clinically meaningful pain developing in roughly 30% of patients. PPSP has physiological, cognitive, and emotional effects and is associated with reduced quality of life. Within the do- main of chronic pain, PPSP is a unique pathological entity that combines nerve injury and inflammatory pro- cesses. PPSP involves peripheral surgical injury of skin, connective tissue, and myofascial-associated tissues, as well as peripheral and central pain sensitization. A growing body of research supports the benefit of MBMT, such as yoga, in treating chronic pain conditions; however, little attention has been devoted specifically to spe- cific needs of individuals with PPSP. This K01 proposes a pilot translational RCT of yoga for PPSP with two research Aims. AIM 1: To design and conduct a pilot feasibility RCT of yoga in individuals with PPSP. This aim contains three specific sections: Aim 1a will adapt a validated yoga program to individuals with PPSP. Aim 1b will assess recruitment, retention, adherence, and acceptability. Aim 1c will assess feasibility of collecting bio- logical and clinical outcomes of PPSP. Exploratory AIM 2: To explore the longitudinal relationship between yoga or an education control group on changes in PPSP-related inflammatory markers and other known psy- chophysical and psychosocial modulators of PPSP. TRAINING. The candidate will achieve a well-defined set of training goals leveraging a resource-rich institutional environment and a cohesive training plan to develop and acquire expertise in: (1) the biological basis of PPSP, (2) the design and conduct of translational clinical trials, and (3) longitudinal analytic experience for translational research. In addition to ongoing mentorship meetings and experiential training through the research plan, the candidate will complete targeted coursework, didactic training and clinical shadowing, present at conferences, and publish in peer-reviewed journals. MEN- TORSHIP. The candidate will be supported by an interdisciplinary mentoring team with requisite expertise in: developing and delivering MBMT interventions for pain conditions, quantitative assessment of pain, analysis of multiple mediators of inflammation, and assessment of patient-centered outcomes relevant to chronic pain con- ditions, and academic career development. The team includes: Drs. Peter Wayne and Kristin Schreiber (co- primary mentors) and Gloria Yeh, Pamela Rist, Robert Saper, Michael Irwin, and Karen Kilgore (co-mentors). IMPACT. The current K01 proposal aligns with NCCIH’s scientific objective to advance the fundamental sci- ence and methods development relevant to MBMT. This K01 will combine didactic, mentoring, and hands-on experience that will facilitate my transition to an independent clinical-translational scientist and lay the founda- tion for broadly studying the used of MBMT for managing and preventing chronic pain conditions.

NIH Research Projects · FY 2026 · 2024-08

This proposal establishes a rigorous and supportive interdisciplinary training program in clinical pain research at Mass General Brigham (MGB) in Boston. The program will support multidisciplinary cohorts of postdoctoral fellows, who will enroll in the program for 2-3 years, with the goal of expanding and enhancing the pool of early career stage investigators who are able to launch and maintain successful careers addressing the nation’s scientific needs in clinical pain research. The program provides protected research time and dedicated training in the context of a unique cohort approach, allowing not only a network of formal mentors, but also peer- mentoring integrated into a rich training experience. In addition to learning through practical engagement in the research process, trainees will undertake coursework tailored to their individual experience, needs, and goals, which can include the completion of graduate certificates or degrees at Harvard University. Required training in responsible conduct of research and research rigor will further establish a solid foundation in clinical research. A national pool of candidates will be recruited, with systematic efforts to establish rigorous academic standards together with a breadth of training backgrounds. The 12 mentors in the program have been selected on the basis of their excellence in clinical pain research and strong mentorship track records. They vary in career stage, background, and expertise, coming from fields beyond those traditionally represented in pain research (e.g., psychology, neurology, neuroscience, psychiatry, epidemiology). These mentors are well-positioned to provide training for eight prioritized pain research content areas: (1) Characterizing and ameliorating group differences in pain outcomes, (2) Understanding and treating chronic overlapping pain conditions, (3) Implementing effective interventions for pain and co-morbidities, (4) Characterizing biopsychosocial mechanisms contributing to persistent pain, (5) Optimizing non-opioid pharmacological treatments for pain, (6) Designing and delivering nonpharmacological interventions for pain, (7) Treating pain across the lifespan, and (8) Preventing the transition from acute to chronic pain. Following participation in the program, trainees will be well-positioned to establish a career of independently-funded clinical pain research and enrich the next generation of research leaders and mentors in pain. The program’s long-term goal is to enhance the number clinical pain researchers with interdisciplinary training and excellent team science skills, in order to promote innovative non-opioid solutions for pain management and improve patient care.

NIH Research Projects · FY 2025 · 2024-08

This proposal establishes a rigorous and supportive interdisciplinary training program in clinical pain research at Mass General Brigham (MGB) in Boston. The program will support multidisciplinary cohorts of postdoctoral fellows, who will enroll in the program for 2-3 years, with the goal of expanding and enhancing the pool of early career stage investigators who are able to launch and maintain successful careers addressing the nation’s scientific needs in clinical pain research. The program provides protected research time and dedicated training in the context of a unique cohort approach, allowing not only a network of formal mentors, but also peer- mentoring integrated into a rich training experience. In addition to learning through practical engagement in the research process, trainees will undertake coursework tailored to their individual experience, needs, and goals, which can include the completion of graduate certificates or degrees at Harvard University. Required training in responsible conduct of research and research rigor will further establish a solid foundation in clinical research. A national pool of candidates will be recruited, with systematic efforts to establish rigorous academic standards together with a breadth of training backgrounds. The 12 mentors in the program have been selected on the basis of their excellence in clinical pain research and strong mentorship track records. They vary in career stage, background, and expertise, coming from fields beyond those traditionally represented in pain research (e.g., psychology, neurology, neuroscience, psychiatry, epidemiology). These mentors are well-positioned to provide training for eight prioritized pain research content areas: (1) Characterizing and ameliorating group differences in pain outcomes, (2) Understanding and treating chronic overlapping pain conditions, (3) Implementing effective interventions for pain and co-morbidities, (4) Characterizing biopsychosocial mechanisms contributing to persistent pain, (5) Optimizing non-opioid pharmacological treatments for pain, (6) Designing and delivering nonpharmacological interventions for pain, (7) Treating pain across the lifespan, and (8) Preventing the transition from acute to chronic pain. Following participation in the program, trainees will be well-positioned to establish a career of independently-funded clinical pain research and enrich the next generation of research leaders and mentors in pain. The program’s long-term goal is to enhance the number clinical pain researchers with interdisciplinary training and excellent team science skills, in order to promote innovative non-opioid solutions for pain management and improve patient care.

NIH Research Projects · FY 2026 · 2024-08

In Alzheimer’s disease (AD), danger signals including dead neurons, dystrophic axons, tau and Ab alter the functional phenotype of microglia from a homeostatic (M0) to a neurodegenerative (MGnD) or DAM (disease associated microglia) phenotype, which in turn drives neuroinflammation and promotes disease. In addition, it is now known that there is a population of disease-associated astrocytes (DAA) in AD which appear at early disease stages and increase in abundance with disease progression. However, the investigation of microglia and astrocytes in AD has been hampered by the lack of understanding their phenotype and function and the lack of therapeutic approaches that can target neuroinflammation associated with these cells. In the mouse model of chronic EAE, we have found that nasally administered anti-CD3 mAb localizes to cervical lymph nodes where it induces IL-10-secreting regulatory T cells (Tregs) that then migrate to the brain and suppress both microglia and astrocyte neuroinflammation. We have recently shown that nasal anti-CD3 improved cognitive deficits in the 3xTg mouse model of AD, an effect independent of amyloid beta deposition, but that was associated with modulation of the microglia phenotype from a MGnD to a M0 gene signature. We have obtained preliminary data in which we found close contact between T cells and microglia in the brains of 3xTg mice treated nasally with anti-CD3 indicating that nasally induced T cells migrate to site of disease in the brain. We hypothesize that induction of Tregs by nasal anti-CD3 will ameliorate disease in the 3xTg AD model by modulating disease associated microglia and astrocytes. Our specific aims are: Aim 1. Investigation of nasal anti-CD3 modulation of microglia and astrocytes in the 3xTg model of AD. We will perform pharmacokinetic (PK) studies to determine the biodistribution as well as optimal dose and regimen of nasal anti-CD3 that modulates microglia and astrocytes. Moreover, mice will be treated with nasal anti-CD3 at different regimens of administration and will undergo behavioral testing. We will 1) quantify Ab and tau levels in the brain; 2) characterize microglia and astrocytes using high throughput RNA sequencing; 3) perform histopathology analysis to measure markers of M0 and MGnD microglia and disease associated astrocytes; 4) investigate microglia function in vivo by measuring phagocytic ability in clearing dead neurons. Aim 2. Investigation of Treg modulation of microglia and astrocytes in the 3xTg model of AD. We will perform fate mapping studies using Foxp3-eGFPCre/ETR2:3xTg-ROSA26Td-TomatoFlox to track Treg cells in the brains of these mice and to determine how long IL-10-secreting Tregs remain active in the brain following nasal anti-CD3 administration. In addition, we will use 3xTg:IL-10Rflox/floxClec7aCreETR2 and 3xTg:IL- 10Rflox/floxGFAPCreETR2 conditional and tamoxifen-induced knockout mice to investigate the role of IL-10 produced by nasal anti-CD3-induced Tregs in modulating microglia and astrocytes. Furthermore, we will use MERFISH to define the spatial location of T cells, microglia and astrocyte subsets and their interactions in 3xTg mice.

NIH Research Projects · FY 2024 · 2024-08

PROJECT SUMMARY/ABSTRACT Aspergillus species are ubiquitous molds that cause clinical disease in hosts with impaired immunity or abnormal mucosal defenses. In particular, individuals with neutropenia or neutrophil defects are particularly susceptible to invasive pulmonary aspergillosis (IPA), a life-threatening respiratory infection with limited treatment options. Neutrophils play a key role in host defenses against this pathogen but little is known about the contribution of these cells beyond their direct microbicidal role. This project examines how neutrophils influence the development of adaptive immunity to Aspergillus in order to better understand mechanisms of host immunity to Aspergillus during neutropenia. In the preliminary data for this proposal, we report that neutrophil- depleted mice have an exaggerated IL-17A response compared to neutrophil-sufficient mice after Aspergillus. We show that following challenge with Aspergillus, IL-17A-producing CD4+ tissue resident memory (TRM) cells persist in the lung up to three weeks after challenge in neutrophil-depleted mice, and that mice that were neutrophil-depleted during a primary Aspergillus challenge are protected from a secondary challenge with Aspergillus when compared to mice that were neutrophil-sufficient during primary challenge. We therefore seek to test the central hypothesis that, in the absence of neutrophils, Aspergillus induces an exuberant IL-17 response which leads to the persistence of IL-17A-producing CD4+ tissue resident memory cells, and that these cells confer protection against subsequent infection with Aspergillus. To test this hypothesis, we plan to pursue the following specific aims: Aim 1: Determine if CD4+ tissue resident memory lymphocytes protect against Aspergillus infection during neutropenia and Aim 2: Identify pathways involved in neutrophil-independent IL- 17A-mediated protection during rechallenge with Aspergillus. The project brings an innovative approach to the study of this infection through the use of a dual challenge model of IPA which mimics repeated exposure to this fungal pathogen in human hosts and the characterization of a novel and previously undescribed protective role for TRM in the setting of neutrophil-depletion. The proposed studies are relevant to public health by defining a new mechanism of host defense against an important human pathogen that should allow for future development of novel therapeutics or preventative strategies.

NIH Research Projects · FY 2026 · 2024-08

Project Summary Reproductive challenges are a growing concern worldwide, marked by a 50% decline in fertility rates over the past 50 years. Approximately 10% of couples face infertility, affecting both men and women. A condition called azoospermia, where semen lacks sperm, impacts up to 20% of infertile men. Azoospermia is categorized into obstructive and non-obstructive types, and assisted reproductive technologies (ART) are often necessary. Microdissection testicular sperm extraction (microTESE) is the preferred surgical method for non-obstructive azoospermia but relies on time-consuming manual examination of samples under an optical microscope. Furthermore, it has been shown that in some cases of non-obstructive azoospermia, rare sperm may exist in the ejaculate, but they can only be identified through laborious manual searches of large-volume samples. Such occurrences can potentially save these individuals from undergoing microTESE surgery. The need for an accurate, reliable, and consistent method for sperm identification and retrieval is crucial for improving the chances of men with azoospermia having biological children. While microfluidics and machine learning have shown potential, they face limitations. Microfluidic solutions suffer from usability and design complexity issues, while machine learning struggles with domain bias. None of the existing systems are suitable for replacing manual sperm examination, highlighting the ongoing need for a clinically viable method to retrieve sperm efficiently and reliably from testicular biopsies, aspirates, and severe factor ejaculated specimens (oligozoospermia/cryptozoospermia). To address these challenges, our proposed approach includes two components: 1. A microwell device designed to digitize cells in ejaculated samples from men with low sperm counts (cryptozoospermia) and microTESE samples. This device creates confined nanowells for efficient sperm localization. 2. A robust deep learning framework tailored to handle diverse and heterogeneous data, enabling swift and precise detection of rare sperm.

NIH Research Projects · FY 2025 · 2024-08

Project Summary/Abstract Research: Psoriatic Arthritis (PsA) is a chronic inflammatory disease that often leads to disability and reduced health-related quality of life. Even with recommended therapy, less than 40% of patients achieve treatment targets such as minimal disease activity (MDA). We previously showed that 73% of patients with PsA experience poor sleep quality compared to 27% of “healthy controls”, and that poor sleep quality is associated with markers of disease activity such as swollen joint counts and pain. Furthermore, our qualitative research suggests a bidirectional relationship between PsA disease activity and sleep quality. Our proposed 12-month longitudinal prospective study will document both objective (i.e., actigraphy) and subjective (i.e., sleep diaries and questionnaires) measures of sleep, novel serum inflammatory markers, and achievement of MDA to test the overarching hypothesis that sleep quality and disease activity are bidirectionally associated, and that these associations are mediated by systemic inflammation. Specifically, we will determine whether (1). poor sleep quality (exposure) reduces the probability of achieving sustained MDA (outcome), (2). moderate-to-high disease activity (exposure) reduces the probability of achieving sustained good sleep quality (outcome), and (3) that systemic inflammation is a mediator in the causal pathway of these associations. The overarching goals of this proposal are to a) better understand the relationship between sleep and PsA disease activity to support the development and testing of interventions to improve sleep health and short and long-term outcomes in PsA and b) provide a training vehicle for Dr. Perez Chada to become an independent investigator. Candidate: Dr. Perez Chada is a dermatologist with current expertise in psoriatic disease and outcome measures research. Environment: Dr. Klerman, the candidate’s primary mentor, and her co-mentors (Dr. Alexis Ogdie, Dr. Monika Haack, and Dr. Joseph Merola) have directly supervised the training of numerous successful junior faculty members. Harvard Medical School and its hospitals and its Division of Sleep Medicine (DSM) offer a rich training environment including resources and expertise. These are available in the NIH-supported DSM Research Training Program in Sleep, Circadian and Respiratory Neurobiology; the Harvard School of Public Health; and Harvard Catalyst. Combined with the expertise available in her mentors’ lab, these opportunities will be important training for Dr. Perez Chada’s career. Career Development: Dr. Perez Chada’s long-term career goal is to become an R01-funded independent, patient-oriented physician-scientist. Dr. Perez Chada’s training will focus on sleep physiology and analysis, advanced biostatical and epidemiological techniques, mechanisms of inflammation, and rheumatology. Completion of the proposed research and career development plan will give the candidate the knowledge and skills to be uniquely positioned to lead research in sleep health and its relationship(s) with skin and musculoskeletal chronic inflammatory disease.

NIH Research Projects · FY 2024 · 2024-08

Caused by advanced cerebral small vessel disease (CSVD), intracerebral hemorrhage (ICH) leaves survivors with a very high risk of incident vascular cognitive impairment and dementia (VCID). While aggressive blood pressure (BP) reduction can reduce the risk of VCID and recurrent stroke, more than 50% of ICH survivors have BP above target, particularly individuals with adverse social determinants of health (SDOH). The parent study, REACH-ICH (R01NS093870), will fill gaps in our knowledge about the role of SDOH after ICH by identifying SDOH-induced environmental risk factors that impact a) VCID after ICH, b) BP treatment resistance, and c) engagement with a program designed to improve treatment of BP. 700 ICH survivors are currently being enrolled through REACH-ICH at multiple centers, divided among White, Black, Hispanic, and Asian participants. Participants are screened with an extensive battery for SDOH exposures, and are followed longitudinally for BP treatment response, cognitive decline, and recurrent stroke. 607 of the participants will receive genome-wide genotyping under the award. Additional analyses building on the REACH-ICH infrastructure could yield biological mechanisms induced by adverse SDOH environments that could be amenable to therapeutic development, at a fraction of the cost of a standalone study. This proposed project, REACH-EpiVCID, will examine epigenetic mediators between SDOH exposures and adverse cognitive outcomes after ICH, as well as BP treatment resistance. Epigenetic changes have already been identified in association with environmental exposures such as air pollution, and have established associations in cardiovascular disease. We will extend these observations to a cohort heavily enriched for CSVD and with an historically high burden of adverse SDOH exposures. REACH-EpiVCID will a) identify epigenetic changes associated with environmental differences induced by SDOH, b) identify epigenetic changes associated with incident VCID and BP treatment resistance after ICH, and c) combine epigenetic associations with DNA genotypes to perform formal mediation analyses and causal inference testing via Mendelian randomization. Epigenetic changes and the mechanistic pathways they highlight are potentially modifiable by small molecules or by behavioral change, and so putative causal associations mediating the effect of SDOH on VCID will represent highly relevant candidates for development of novel treatments to prevent VCID and improve BP treatment resistance after ICH. Further, because epigenetic mediators between SDOH and VCID are likely to be initiated by adverse environmental exposures, treatment targets arising from this approach stand to have the greatest benefits in marginalized populations at greatest risk for these exposures, promoting health equity through biologically informed treatments as strategies to ameliorate the structural contributors to SDOH continue to evolve with time.

NIH Research Projects · FY 2025 · 2024-08

Project Summary/Abstract Calcific Aortic Valve Disease (CAVD) will affect 3% of people over the age of 75. CAVD disease progression is characterized by an active deposition of calcific noduli and extracellular matrix proteins. This excessive deposition results in valvular thickening, outflow tract narrowing, restricted blood flow, left ventricular hypertrophy, and eventual heart failure. Despite the clinical significance of this disease, patients must “watch and wait” until surgical AV replacement and repair is necessary, as currently no pharmacotherapeutics exist. This proposal focuses on identifying novel epigenetic mechanisms underlying calcific aortic valve disease progress and pathophysiology. For preliminary investigation on the role of epigenomic regulators in valve calcification, we re-mined proteomic datasets to specifically probe differential abundance of epigenetic factors – that is, proteins involved in histone post-translational modification reading, writing, and erasing. The preliminary data presented in this proposal shows that enzymes responsible for histone regulation are differentially abundant in valvular tissue as a function of disease stage, structural localization within the valve leaflet, as well as within VIC cultures as a function of calcification induction media (inorganic vs. organic phosphate media). However, the dataset mined was not exhaustive in identification as it was untargeted. Additionally, our preliminary data did not investigate the regulatory role epigenetics plays in downstream translational and post-translational signaling required for cell-cell, cell-matrix, and cell-vesicle mediated signaling. The proposed research capitalizes on an ever-expanding cohort of clinically defined human adult CAVD aortic valve tissue, as well as an extensive biobank of valvular interstitial cells isolated from human donors. It is our central hypothesis that there are unique histone modifications that contribute to pathological development of calcification in human aortic valves. Aim 1 will use novel mass spectrometry approaches to define the histone code of CAVD along with corresponding transcriptional regulation via Chromatin Immunoprecipitation sequencing. Aim 2 will determine cell-mediated spatially localized translational targets downstream of epigenetic regulation, utilizing multi-modal histopathological imaging, laser capture microdissection, and low-input proteomic strategies. Aim 3 will investigate the role of epigenomic modifications on microenvironment signaling mediated by N-linked glycosylation. By mapping the histone code of aortic valve calcification and identifying both upstream epigenetic regulators and downstream transcriptional, translational, and post-translational targets of this epigenetic regulation, we aim to identify potential pharmacotherapeutic targets that may halt progression of CAVD. These studies will be conducted by Dr. Clift under the mentorship of Dr. Elena Aikawa, a pioneer in cardiovascular systems biology, as well as an advisory committee dedicated to proposed research and trainee. By utilizing this mentorship and professional development via the MOSAIC UE5, Dr. Clift is primed for successful independence.

NIH Research Projects · FY 2025 · 2024-08

Project Summary Gynecologic malignancies accounted for over 114,810 new cancer cases and approximately 35,640 deaths in 2023 in the United States. Brachytherapy has been used to treat locally advanced cervical and endometrial cancers since the early 20th century and is now part of the standard of care. Brachytherapy involves the precise placement of short-range radioactive sources near or in direct contact with the tumor through thin catheters, enabling high radiation doses in the target volume with rapid fall-off to protect adjacent normal structures. Today, high-dose-rate (HDR) brachytherapy is commonly employed along with a computer-controlled remote afterloading system, which allows accurate control of radiation dose for each catheter by adjusting the “dwell time.” Though computer-controlled afterloading systems are widespread, the quality of brachytherapy is still limited by suboptimal catheter placement. Clinicians often struggle to properly deploy the catheters in the target volume because of the deviation of the catheters from the intended path during insertion and lack of quantitative catheter position feedback or the dosimetric consequences resulting from the current catheter locations. Intraoperative imaging, either computed tomography (CT) or magnetic resonance (MR), potentially provides such feedback and allows for “adaptive catheter placement,” where the clinician adjusts catheter location until optimal dosimetry is achieved. However, adaptive catheter placement is not practical in the current form because it requires iterative implantation and imaging; each iteration involves positioning of the patient for imaging and catheter placement and moving of the clinician between the imaging room and the control room. To enable adaptive catheter placement in a wide range of clinical settings, we will develop a catheter placement manipulator system that combines (1) a state-of-the-art fiber-optic shape-sensing stylet to obtain real-time quantitative measurement of the catheter trajectories in the patient, and (2) a teleoperated catheter placement manipulator to shorten the turnaround time for catheter placement and evaluation, (3) a visualization framework that provides quantitative measures of a catheter’s deviation from its intended trajectory and real-time evaluation of the consequences to the achievable radiation dose distribution. We hypothesize that the combination of real- time catheter trajectory digitization and quick catheter insertion will allow adaptive catheter placement, where the catheter locations are optimized through frequent iteration of the plan-insert-check cycle with real-time quantitative dosimetry feedback, leading to optimal radiation dose distribution. We will pursue the following specific aims: (Aim 1) Develop a fiber-optic shape-sensing stylet for real-time catheter tracking to achieve real- time tracking and prediction of the catheter trajectory for real-time feedback; (Aim 2) Develop a teleoperated catheter insertion manipulator for quick catheter placement to achieve a shorter turnaround time for the frequent plan-insert-check cycle; (Aim 3) Develop, optimize, and validate the system for teleoperated adaptive catheter placement to test the impact of teleoperated adaptive catheter placement for optimal dose distribution.

NIH Research Projects · FY 2025 · 2024-08

Project Summary Buprenorphine reduces overdose mortality by up to 70%, making it one of the most critical interventions to combat the opioid overdose crisis. With the increasing prevalence of illicit fentanyl, patients with opioid use disorder (OUD) attempting to initiate buprenorphine now routinely report experiencing buprenorphine precipitated opioid withdrawal (BPOW) despite waiting for withdrawal symptoms to first emerge. In response, clinicians today are increasingly recommending a novel strategy called “micro-dosing” or “low-dose” buprenorphine induction (LDBI), where a dose significantly lower than SL buprenorphine (SL-BUP) 4mg is administered before the emergence of any withdrawal symptoms. The requirements for a successful LDBI appear to be the sufficiently low initial dose, the up-titration of the SL-BUP dose over several days, and continuation of the full agonist opioid during the induction. However, in the emergency department (ED) or outpatient settings where individuals with OUD routinely seek care, the slow up-titration over several days and the continuation of the full agonist are difficult to implement. As such, research is urgently needed for buprenorphine induction strategies that can be easily and safely implemented in the ED or outpatient settings. One possible solution is to utilize extended-release buprenorphine (XR-BUP). The “standard” approach to initiating XR-BUP is to administer the first injection after 7 days of SL-BUP. However, XR-BUP is increasingly used in a “rapid” induction protocol, where XR-BUP is given one hour after the successful administration of even a single dose of SL-BUP. Furthermore, an ongoing NIDA-funded trial is demonstrating that XR-BUP can be “directly” initiated without first administering SL-BUP at all, where XR-BUP is administered after the emergence of opioid withdrawal. However, due to the slow rise in the serum buprenorphine levels, XR-BUP may function as a LDBI strategy on its own, allowing for XR-BUP to be administered before waiting for opioid withdrawal to emerge, without using any SL-BUP or concurrent full agonists opioids. If successful, this strategy could facilitate buprenorphine initiation in the ED and outpatient settings. Unfortunately, no prior research has evaluated whether this novel LDBI strategy using XR-BUP is safe. To fill this need, we propose to conduct a double-blind, randomized trial in an inpatient setting with individuals with OUD (n=30). Participants will be randomized to receive XR-BUP 16mg (n=10), 24mg (n=10) or 32mg (n=10) and remain in a controlled inpatient setting for 5 days. The primary aim is to determine the safety of using XR-BUP as a LDBI strategy. The secondary aim is to conduct pharmacokinetic analyses of buprenorphine, norbuprenorphine and metabolites. If successful, this research could facilitate the development of a safe approach to initiating buprenorphine treatment without the need to wait for the emergence of opioid withdrawal, require the need for SL-BUP, or continue the full agonist opioid. Results from this trial will then lay the groundwork for an adequately powered pragmatic randomized controlled trial utilizing XR-BUP as a LDBI strategy in the ED and outpatient setting.

NIH Research Projects · FY 2025 · 2024-08

Summary: ABO(H) blood group antigens and corresponding anti-ABO(H) alloantibodies were discovered over a century ago as the first polymorphisms in the human population and continue to be the most common immunological barrier to transfusion and transplantation. However, despite the fundamental nature of this discovery, very little is surprisingly known regarding the factors that govern anti-ABO(H) antibody formation or the fine details of the ABO(H) alloantigen targets responsible for hemolytic transfusion reactions (HTRs). As a result, current approaches designed to detect ABO(H) antigens and anti-ABO(H) antibodies largely rely on the same agglutination strategy leveraged by Landsteiner over 122 years ago. While ABO(H) incompatible RBC transfusion can result in a hemolytic transfusion reaction (HTR), only half of patients who receive ABO(H) incompatible RBCs experience this outcome. However, the factors that contribute to variable ABO(H) incompatible HTRs remain largely unknown. This limitation in our understanding is a direct consequence of the complexity of the post-translational modifications that comprise ABO(H) antigens and a historical limitation in the tools needed to study both ABO(H) antigens and the antibodies that develop against them. While a variety of highly novel tools have begun to revolutionize the field of glycosciences – the study of carbohydrate modifications – these tools have not been as thoroughly applied to perhaps the most common and arguably most clinically significant carbohydrate structures within the human population – ABO(H) blood group antigens. Fundamental questions surrounding anti-ABO(H) antibodies and their target antigens often require disparate areas of expertise, including glycosciences, immunology, hematology and microbiology, which has directly limited the study of this foundational discovery in transfusion medicine. To overcome this challenge, we have assembled a highly collaborative and integrated team of physicians and scientists with expertise in these fields. The combined collaborative history of the Project leaders (>50 papers and >15 years of collaboration) and the use of newly developed models and tools specifically designed to define factors that govern anti-ABO(H) antibody development and the specific ABO(H) targets on RBCs responsible for HTRs. In doing so, this program project grant (PPG) will provide a unique opportunity to define fundamental features of anti-ABO(H) antibody development and the ABO(H) targets that result in HTRs that have remained incompletely understood for over a century. This will be accomplished through 3 distinct Projects and the support of complementary Cores. Project 1: Examining the impact of microbial dynamics on B cells responsible for anti-blood group antibody formation (Leader: Stowell). Project 2: Convergence of innate immunity and microbial communities in the regulation of anti-blood group antibody development (Leader: Arthur). Project 3. Defining the distinct antigen targets and antibody specificities that govern ABO(H) RBC incompatibility (Leader: Cummings). By leveraging these Projects and Cores, this PPG will answer fundamental questions in transfusion medicine.