Brigham And Women'S Hospital

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY The digit tips of many mammals, including humans, innately regenerate following amputation. Multi-tissue digit tip regeneration requires coordinated proliferation and interaction of many cell types in a structure termed the ‘blastema’. Significant progress has been made in defining the origin and cellular composition of the mouse digit tip blastema, but there is a gap in our understanding of how the heterogeneous blastema becomes a correctly organized and shaped composite tissue. This proposal focuses on defining the molecular mechanism(s) of patterning the regenerating mouse digit tip. In general, broad similarities between the regenerative blastema and the embryonic limb bud underlie the hypothesis that gene networks required for embryonic limb patterning are re-deployed in the regenerative blastema. While studies support this hypothesis in some non-mammalian models of limb regeneration, our data demonstrate that this is not true for dorsal- ventral patterning during mouse digit tip regeneration. This suggests that mammalian and non-mammalian limb-derived blastemas may employ different mechanisms for re-establishing limb tissue morphology. We propose that the mammalian digit tip blastema utilizes non-developmental patterning mechanisms and receives molecular patterning cues from the nail epithelium. In this proposal, we focus on the dorsal-ventral and proximal-distal anatomical axes; we aim to determine the patterning mechanism(s) in the mouse digit tip blastema and to determine how manipulation of these pathways during regeneration impacts morphology. Toward this, we will model and molecularly define proximal-distal patterning genes using computational and genetic approaches. We will also use grafting studies to determine the tissue patterning autonomy of the digit tip blastema. Finally, we will visualize putative dorsal-ventral electrochemical gradients in the blastema and perform gain and loss of function studies to manipulate these signals. Collectively, the data generated from this project will reveal patterning mechanisms for mammalian digit tip regeneration and highlight how this may be fundamentally different from non-mammalian limb regeneration models. This research will ultimately inform blastema induction efforts for mammalian full limb amputations because exogenous patterning information may need to be provided.

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY/ABSTRACT Chronic obstructive pulmonary disease (COPD) is projected to become the third leading cause of death globally by 2030. While smoking is a primary risk factor, a growing body of epidemiological research points to the role of nutrition in modulating the risk for COPD and COPD-related manifestations, including acute COPD exacerbations (AE-COPD). However, the mechanisms underlying the impact of dietary components on COPD and AE-COPD are not well understood. The primary objective of this proposal is to develop and implement a new set of machine learning and network science-based methods specifically designed to explore the impact of dietary xenobiotics (DXs) on AE-COPD. DXs are small molecules directly derived from food digestion, absorption, and related metabolism, captured by untargeted metabolomics. This study ultimately aims to gain a precise molecular understanding of the mechanisms driving the systemic effects of diet on AE-COPD. The research design involves developing new methods that leverage the multi-omics data from COPDGene and SPIROMICS, including longitudinal clinical and molecular profiling of smokers with and without COPD. First, we will develop ensemble learning techniques using plasma metabolomic data to identify a DX signature predictive of AE-COPD. Multiple statistical strategies will be integrated to rank and group DXs based on their ability to predict AE- COPD frequency and severity (Aim 1). Second, we will combine metric learning algorithms and spectral theory to quantify the coordinated effect of DXs and circulating proteins on AE-COPD. The goal is to capture the interplay between proteins and DXs in discerning exacerbation-prone phenotypes (Aim 2). Third, we will utilize pre-trained deep learning models and network science to implicate potential mechanisms of action of DXs on AE-COPD. A pipeline will be designed to annotate and predict DXs’ protein targets and assess their proximity on the interactome to different subregions of the COPD disease module, to key inflammatory proteins and epigenetic modifiers (Aim 3). This K25 award will facilitate Dr. Menichetti’s comprehensive training in longitudinal analysis, pulmonology, inflammatory processes, and integrative omics methods. Addressing these training gaps will enhance her ability to develop reliable and translationally relevant methods establishing the foundation for personalized dietary interventions based on distinct COPD endotypes. Dr. Menichetti has crafted a detailed training plan and assembled a mentoring team with complementary expertise, ensuring she will receive the necessary guidance and support to successfully complete her proposal. This K25 will ultimately enable Dr. Menichetti’s independent career in the methodological foundation of Precision Nutrition for lung disease.

NIH Research Projects · FY 2026 · 2024-04

The goal of this proposal is to validate glycated CD59 (GCD59) in human urine as a novel diabetes biomarker. This validation will be conducted across several hyperglycemic conditions including pregnancy-induced glucose intolerance/gestational diabetes. The proposal is highly translational and addresses major Public Health priorities because: 1) women with pregnancy-induced glucose intolerance and their fetus are at an increased risk of adverse clinical outcomes, 2) the frequency of pregnancy-induced glucose intolerance is increasing at alarming rates, 3) appropriate treatment reduces the associated risks for mother and newborn, and 4) glucose load tests, currently the gold standard identifying pregnancy-induced glucose intolerance, are expensive, time consuming, unpleasant, and have poor reproducibility on repeat testing. These facts highlight why a simpler, shorter, easier-to-use, cost-effective, sensitive and specific test would be a much better tool to screen for pregnancy-induced glucose intolerance. The applicants have 1) discovered that human CD59 is inactivated by glycation, 2) provided evidence for a link between the complement system and the complications of diabetes, and 3) developed key reagents and established specific and sensitive assays that allows quantification of GCD59 in blood. With this assay, we have conducted several published studies including ≈4,000 subjects with diabetes, pre-diabetes or gestational diabetes. Based on the robust preliminary data from these studies and available resources, we now propose the highly focused aim of assessing the clinical utility of urine GCD59 as a simple, easy-to-use test for screening and diagnosis of diabetes, pre-diabetes and gestational diabetes. All necessary tools and expertise to accomplish our aims are available in the laboratory of the applicant and expert collaborators, including monoclonal antibodies specific for GCD59 and assay calibrators, diagnostic tools, equipment and expertise necessary to conduct all studies proposed in the application. Through the Crimson Biomaterials Collection Core Facility we have access to urine and plasma samples from ≈ 12,000 pregnant women seeking prenatal care every year at the two main Partner’s Healthcare Hospitals. Successful accomplishment of our aim will provide a clinically useful biomarker that could simplify screening, diagnosis and management of conditions characterized by hyperglycemia.

NIH Research Projects · FY 2026 · 2024-04

Project Summary Pulmonary fibrosis (PF) is one of the most common forms of interstitial lung disease (ILD) and is often progressive leading to loss of lung function and death. Given that anti-fibrotic therapy can reduce progression, even in patients with less severe disease, there is more urgency to detect early stages of PF. Dr. Rose’s research group has helped to define imaging patterns, called interstitial lung abnormalities (ILA), that can help to predict early stages of PF. The fact that ILA is so much more prevalent than IPF, however, compels us to identify the subsets of ILA most likely to experience adverse outcomes. Dr. Rose’s initial work in the COPDGene cohort demonstrated that a combination of advanced imaging findings and pulmonary function identifies a high-risk group of ILA. The optimal method for predicting adverse outcomes, however, remains unclear and replication in other populations is warranted. Furthermore, novel biomarkers are needed, and the mechanisms of severity in this disease need to be better understood. The overall goal of this project is to identify the best method for the risk-stratification of ILA in order to identify a subset that may benefit from early intervention. In the first aim, Dr. Rose will assess the ability of clinical variables to predict a range of longitudinal outcomes and identify the optimal set of clinical criteria that helps to classify high-risk ILA. In the second aim, he will utilize proteomic measures to evaluate plasma proteins and multi-protein models that help to risk-stratify ILA. Finally in the third aim, he will explore genetic predictors of the proteins most significantly associated with high-risk ILA. This work will be performed in the Division of Pulmonary and Critical Care Medicine, at Brigham and Women’s Hospital (BWH), a core teaching hospital of Harvard Medical School. Dr. Rose will perform this work under the mentorship of Dr. Hunninghake, an expert in the field of early IPF characterization and Dr. Raby, an expert in genetic epidemiology. With the guidance of his mentors and scientific advisory committee, Dr. Rose has developed a comprehensive five-year training program to develop the skills needed to complete the project and become an independent investigator with expertise in imaging characterization, machine-learning techniques, complex multi-omic analyses, and their integration. Dr. Rose is dedicated to a career in academic medicine. His goal is to become a physician-investigator using the knowledge and skills gained during this project to better our understanding of adverse outcomes in pulmonary fibrosis and the biologic processes that lead to progressive disease, in a way that will ultimately benefit the patients suffering from this disease.

NIH Research Projects · FY 2026 · 2024-04

Project Summary/Abstract Traumatic brain injury (TBI) affects millions of people each year who recover to widely disparate levels of independent function. It is not currently possible to reliably predict who will remain functionally dependent on caregivers. Most patients assessed with existing models are assigned an intermediate likelihood of recovery, remaining in a prognostic ‘grey area’. Accurate prediction is important, because withdrawal of life sustaining therapy based on perceived prognosis is the leading cause of death after TBI. Though advances in brain imaging have enabled precise localization of focal brain injuries, injury location is not incorporated into existing TBI prognostic models. This is because it is not known whether dependency results from injury to specific brain structures or networks. This knowledge gap creates ongoing heterogeneity in clinical practice and limits the development and evaluation of targeted therapeutics. This K23 award addresses this knowledge gap, using multi-modality neuroimaging to identify brain structures, connections and networks that produce dependency when disrupted, and testing whether injury location improves prognostication after TBI. The principal investigator, Dr. Samuel B. Snider, is a Neurocritical Care Neurologist at Brigham and Women’s Hospital (BWH), whose goal is to become a translational neuroscientist using advanced imaging techniques to better understand recovery mechanisms and predict outcomes after acute brain injuries. Dr. Snider has an established early career track record in advanced MRI, and in measuring and predicting TBI outcomes. His preliminary data demonstrate the feasibility of mapping focal traumatic brain injury with CT and MRI at the scale required for this project. Through novel analysis of three existing datasets and one prospectively enrolling TBI study at BWH, Dr. Snider will test whether the locations of hemorrhagic contusions on CT scans (1), axonal injury on diffusion MRI (2a) and functional network disruption on resting state functional MRI (2b) independently improve the prediction of functional dependency after moderate or severe TBI. Using multiple sources to create one of the largest TBI imaging datasets ever assembled, this project will generate novel insights into mechanisms of recovery from brain injury and improve existing prognostic models. The mentored research and structured training in multi-center data harmonization and analysis, resting-state functional MRI, and prospective clinical data collection will provide Dr. Snider with the skills, experience and preliminary data needed to submit an NIH R01 validating the first imaging-based TBI prognostic model. His career development plan utilizes the resources of the world-class Harvard Medical School training environment, bringing together a diverse and multidisciplinary team of mentors and collaborators centered at the BWH. Under the guidance of primary mentor Dr. Michael Fox, co-mentors Drs. Brian Edlow and Nancy Temkin, and advisors Drs. Alexandra Golby, Yogesh Rathi, and Sonia Jain, Dr. Snider will find the support necessary to develop an independent research program translating better prognostic models into better outcomes for patients with brain injuries.

NIH Research Projects · FY 2026 · 2024-04

Project Summary Effective treatments for COPD are disappointingly limited because the subtypes of this complex disease are not well defined. COPD may be initiated by susceptibility of different cell types to pollutants, including epithelial cells, endothelial cells, fibroblasts, dendritic cells, T cells, or any other cell type. We have shown that primary human airway epithelial cells (HAECs) with an arginine (HAECArg ) compared with a proline ( HAECPro) at locus 72 of the p53 protein, express higher mucin levels and that the same is found in nasal brushings from humans. Single- cell RNA sequencing showed that differentiated HAECArg show increased numbers of mucous cells while HAECPro cultures show squamous cells. Therefore, genetic variants may cause susceptibility depending on the cell type and the type of injury that initiates the chronic inflammatory condition. To begin investigating this possibility, we generated whole genome genotyping and bulk RNA sequence data from HAECs of 185 individuals cultured as basal cells. We identified 1,014 genes that harbored at least one expression quantitative trait locus (eQTL) and 16 distinct loci containing known COPD-associated genetic variants including a region near MUTYH. Based on these findings, we propose to test the hypotheses that multiple genetic loci associated with COPD harbor eQTLs or protein QTLs that are active in airway epithelia exposed to cigarette smoke (CS). Colocalization analysis of these QTLs with COPD genome-wide association study (GWAS) results will identify airway-specific genes that initiate COPD development. Aim 1 will identify CS-responsive genes and pathways and link eQTLs and protein QTLs to COPD-associated genetic variants, with MUTYH being the first identified gene to investigate. Aim 2 will determine the role of the Tp53 variants in airway epithelial differentiation. Isogenic HAECArg and HAECPro cultures will be compared for cell types by single cell RNA sequencing and investigate whether the p53Arg or p53Pro variants facilitate a mucous or squamous differentiation process depending on culture conditions. Mechanisms underlying the p53Arg-mediated expression of mucin genes will be elucidated. These studies will help define pathways that characterize a specific COPD subtype and provide biomarkers for early diagnosis, patient stratification. The data from HAEC185 will also provide a resource for future studies on any chronic disease that originates from susceptible airway epithelial cells.

NIH Research Projects · FY 2026 · 2024-04

Macrophage migration inhibitory factor (MIF) is one of the first cytokines reported nearly 60 years ago. Yet, it took 40 years to find out its receptor CD74. CD74 is also called invariant chain that chaperones MHC-II maturation from ER to endosome/lysosome where CD74 gets degraded by cathepsins. CD74 does not contain a signaling component on its short cytoplasmic tail. This nature of CD74 inspired later discoveries that CD74 does not work alone, but forms complexes with CD44 and chemokine receptors CXCR2 and CXCR4. Since we first reported a direct role for MIF in atherosclerosis, and suggested MIF activity in abdominal aortic aneurysms (AAA) at the time of the CD74 report, we have also started searching for novel MIF receptors. Using mouse aortic smooth muscle cells (SMCs) and recombinant MIF as bait, we discovered a 110-kDa MIF-binding protein: chloride channel accessory protein-1 (CLCA-1). We initially thought it might be a new MIF receptor. CLCA1 is a Ca2+-activated chloride channel (CaCC) expressed in airway epithelial cells, where it lowers the energy barriers for ion transportation as a mechanism to enhance mucin expression and respiratory dysfunction. CLCA1 contains three domains: the CLCA-N domain located on the N-terminus with metallo-hydrolase activity that cleaves CLCA1 into two pieces, the 75-kDa N-terminal fragment and 35-kDa C-terminal fragment, the von Willebrand factor type A (vWA) domain within the 75-kDa fragment to engage the channel to increase Ca2+- dependent Cl– current, and the fibronectin type-III (FN-III) domain within the 35-kDa C-terminal fragment with untested function. Although CLCA1 is not a MIF receptor as we wished, we found that the 35-kDa fragment binds to MIF and acts as a decoy inhibitor that blocks MIF-induced macrophage expression of cytokines and chemokines. We did not detect significant correlations between plasma 35-kDa CLCA1 fragments with AAA size or AAA growth rate from an AAA screening trial, but plasma 75-kDa fragment levels correlated strongly with AAA size and growth rate. We revealed deficiency of the 35-kDa fragment in human and mouse AAA lesions. This fragment is expressed in ECs and SMCs from healthy or diseased aortas. In contrast, the 75-kDa fragment is expressed mainly in macrophages. In CaPO4 injury-induced AAA, MIF deficiency (Mif–/–) reduced AAA growth, but CLCA1 deficiency (Clca1–/–) expedited AAA growth. The increased AAA growth in Clca1–/– mice was fully muted in the absence of MIF (Clca1–/–Mif–/–). CLCA1 with mutations at the cleavage site or the protease active site failed to undergo self-cleavage. CLCA1 knock-in at the cleavage site or protease active site increased AAA growth. We hypothesize that, instead of serving as a MIF receptor, CLCA1 uses its C- terminal 35-kDa FN-III domain-containing fragment to act as a MIF decoy inhibitor that binds to MIF, blocks MIF-induced activation of aortic wall inflammatory and vascular cells, and slows AAA growth. We proposed two Aims to examine the decoy inhibitory role of CLCA1 and its 35-kDa FN-III fragment in MIF-induced AAA and to explore CLCA1 fragment-mediated decoy inhibition of MIF activity on macrophages and vascular cells.

NIH Research Projects · FY 2026 · 2024-03

Summary: While ABO(H) blood group antigens were the first human polymorphisms described and corresponding anti-ABO(H) antibodies continue to be the most common immunological barrier to transfusion and transplantation, the factors responsible for anti-ABO(H) antibody development remain relatively unknown. In order to overcome barriers that result from anti-ABO(H) antibody formation, key processes that drive anti-ABO(H) antibody development need to be defined. Our long-term goal is to define the factors that regulate naturally occurring anti-ABO(H) antibody formation. Our central hypothesis is that exposure to microbes that decorate themselves with ABO(H) blood group-like antigens drives innate-like B1 B cells to produce naturally occurring anti-ABO(H) antibodies. Our hypothesis is formulated on the basis of our recent discovery that microbes that decorate themselves with carbohydrate structures that mimic blood group antigens stimulate the formation of anti-blood group antibodies capable of causing hemolytic transfusion reactions (HTRs). Our data demonstrate that anti-ABO(H) antibodies isolated from patients display unique specificity for distinct types of ABO(H) antigens and when examined against microbial glycans isolated and presented on a microbial glycan microarray, engage unique microbial determinants, strongly suggesting that microbial glycans may shape an individual's anti-ABO(H) antibody response. However, as ABO(H) blood group antigens are carbohydrate structures largely confined to humans, preclinical models capable of formally testing this have not been available. To overcome this limitation, we developed a preclinical model that recapitulates key features of naturally occurring anti-blood group antibody formation. Knocking out the enzyme required for the synthesis of the murine blood group B-like antigen (murine B or Bm), we generated blood group O-like (murine O or Om) mice that spontaneously develop varying levels of anti-Bm antibodies capable of causing Bm RBC HTRs following transfusion. Sorting and culturing anti-Bm reactive microbiota identified a strain of Klebsiella pneumonia that specifically expresses the Bm antigen, providing a possible link between the microbiota and anti-Bm antibody development. As blood group Om mice and O individuals possess innate-like B1 B cells with blood group specificity, these collective data suggest that microbial stimulation of B1 B cells drives the formation of anti-blood group antibodies capable of causing HTRs. To test this hypothesis, we will pursue the following specific aims: Aim 1. Define the role of B1 B cells and anti-blood group antibody reactive microbiota in the development of anti-ABO(H) antibodies capable of causing HTRs. Aim 2. Define the requirement for B1 B cells in microbiota-induced anti-Bm antibody formation using a preclinical model. These aims will not only define the impact of the microbiota on the development of anti-blood group antibodies, but provide a rich training opportunity for me to weld my previous training in transfusion immunology with new training in glycobiology to define the governing factors that regulate the development of the most common immunological barrier in not only transfusion, but also transplantation.

NIH Research Projects · FY 2026 · 2024-03

PROJECT SUMMARY Asthma exacerbations are a major cause of disease morbidity and lead to progressive loss of lung function, airway remodeling, and enhanced disease severity. Their associated economic burden is substantial, totaling approximately 15 million outpatient visits, 2 million emergency room visits, and 500,000 hospitalizations each year in the United States alone. Predisposition to asthma exacerbations is influenced by genetics and triggered by environmental exposures that result in a dysregulated immune response. Viral and bacterial infections are the leading risk factors for asthma exacerbations. While a few virulent species have been well-studied, a comprehensive assessment across a range of viral and bacterial stimuli, as well as their interactions with multiple omics, to understand how these are linked to individual susceptibility has not been performed. Recent technological advances in Phage ImmunoPrecipitation Sequencing (PhIP-Seq) have facilitated a high- throughput, cost-effective evaluation of global virulent exposures, enabling for the first the estimation of cumulative exposures in the context of asthma exacerbations. As such, our overarching objective is to identify the impact of prior viral, bacterial, and other virulent exposures on asthma exacerbations and severity by employing PhIP-Seq profiling coupled with multiple omics (genetics, epigenetics, and metabolomics). Our overall hypothesis is viral and bacterial exposures interact with host genetics and downstream omics differentially across individuals; these relationships are relevant to disease exacerbations and will differ across the life course. We will first characterize lifetime virulent exposures through PhIP-Seq profiling, then identify exposures and multi- omic variants associated with asthma exacerbations- as well as interactions between these complementary omic layers- and finally, construct universal and personalized predictive models of the impact of these exposures on asthma severity through a cooperative learning methodological approach. We will leverage four well- characterized child and adult asthma cohorts, totaling 4,000 individuals, with existing genetic, epigenetic, and metabolomic data, and apply novel approaches to data integration. The demographic diversity of these cohorts allows us to interrogate both life course effects, as well as sex as a biological variable and differences by race and ethnicity. These results hold unprecedented potential to enlighten the complex relationships between virulent exposures, omic profiles, and asthma exacerbations, providing novel avenues for more personalized therapeutic targets and management strategies.

NIH Research Projects · FY 2026 · 2024-03

The obesity epidemic will affect 1 in 2 adults in the US by 2030. While several medical and surgical weight reduction strategies are available, with newer, transformative medications recently approved, little is known about their safety and effectiveness in clinical practice. Real-world evidence (RWE) studies based on routinely generated health insurance claims from millions of patients have the size and the breadth of data capture needed to provide critical information on the safety and effectiveness of weight loss strategies in clinical practice, complementing evidence from RCTs. In this setting, however, the adiposity measure that guides weight loss treatment, i.e., body mass index (BMI), although captured with high specificity, is often missing. Yet, BMI information is necessary to accurately identify study populations, adjust for confounding, and assess effect modification on a large scale in claims data. Similarly, confounding and other biases may be present in RWE studies. Thus, identifying methodological approaches that increase the validity of RWE studies comparing weight loss strategies is critical to generate unbiased findings that can validly address questions not answered or answerable by RCTs. Our overarching goal is to develop, implement, and test approaches to produce large scale, high-quality RWE on the comparative safety and effectiveness of medical and surgical weight loss strategies to complement RCTs. To achieve this goal, we will leverage (1) the Nurses’ Health Studies (NHS) I and II and the Health Professionals Follow-up Study (HPFS), large longitudinal cohort studies with rich lifestyle and dietary data, linked with their insurance claims data, and (2) large U.S. national (federal and commercial) claims databases, resulting in a heretofore untapped new data infrastructure. We will improve existing algorithms to measure BMI in national claims data using primary data from longitudinal cohort studies (NHS I and II, and HPFS) linked with their claims data (Aim 1); assess the success of approaches to reduce confounding, including comparator choice and propensity score adjustment, with respect to measures of achieved comparability between medical and surgical weight loss strategies identified in claims-based RWE studies with regard to information only measured in the linked data (Aim 2); evaluate when RWE studies of weight reduction strategies emulating target trials provide causal conclusions (Aim 3); and fill gaps in the RCT evidence in targeted questions regarding the safety and effectiveness of medical and surgical weight reduction strategies in clinical practice (Aim 4). This work will develop more accurate, methodologically rigorous approaches to conduct RWE studies of weight reduction strategies using large scale, real-world data and will create an invaluable infrastructure to conduct studies on the comparative safety and effectiveness of various cardiometabolic treatments.

NIH Research Projects · FY 2026 · 2024-03

PROJECT SUMMARY The plasticity of cell identity is evident through nuclear transfer and transcription factor (TF)-mediated reprogramming. Despite this, current reprogramming methods often yield developmentally immature and heterogeneous cell populations and therefore are unsuitable for therapeutic application or disease modeling. We seek to address the fundamental questions of why direct reprogramming is inefficient, representing a critical gap in knowledge that will be widely applicable across many cell fate engineering strategies and has broader significance for understanding how cell identity is regulated. Over the past five years of NIGMS- supported research, we have developed and applied innovative single-cell multiomic lineage tracing and novel computational technologies to dissect the mechanisms of pioneer TF-mediated reprogramming. Our proposed research builds on this work, focusing on two primary objectives: 1. Current evidence supports the hypothesis that successful reprogramming events arise from rare 'reprogramming permissive' cell types or states in which normally inaccessible target genes are engaged by ectopic TFs to drive fate change. However, the lack of understanding about the origins of reprogramming presents a significant challenge in characterizing reprogramming permissive states. We will develop and apply multiomic lineage tracing to identify the origins of successfully reprogrammed cells across various cell fate conversion strategies. Elucidating reprogramming initiation mechanisms will identify new avenues to enhance reprogramming efficiency, uncovering common and cell-type specific regulation of cell identity. 2. The identification of new, more effective reprogramming cocktails represents a current gap in cell engineering. We hypothesize that expanded cocktails of precisely delivered TFs targeting the gene regulatory networks controlling terminal cell identity will more faithfully reprogram fate. However, it is currently experimentally and computationally intractable to predict these cocktails de novo. We will use cell fusion in combination with our genomic technologies to empirically deconstruct gene regulation, providing unique insights into efficient and accurate reprogramming, informing the development of novel TF cocktails. The outcomes of this research will have significant impacts on multiple levels: a) It will facilitate improved conversion efficiency and fidelity across different cell engineering strategies, overcoming a current barrier in regenerative medicine; b) The experimental manipulation of cell fate offers a valuable model system to deconstruct and model dynamic changes in cell identity. Our study of diverse reprogramming strategies will uncover general and cell type-specific rules for cell fate specification and maintenance, providing broad biological relevance beyond cell engineering; c) We will continue to develop our innovative genomic technologies, which provide insight into the regulation of cell identity across diverse cell biology paradigms.

NIH Research Projects · FY 2026 · 2024-03

Project Summary Although neuronavigation systems are of crucial assistance during cerebrovascular surgery, they do not integrate hemodynamics information needed to treat complex cerebrovascular malformations. The present project aims at developing an Augmented Reality (AR) neuronavigation tool that will enable the visualization of cerebral hemodynamics information in the surgical view. Our long-term goal is to contribute toward the development and clinical adoption of visualization tools that allow for safe and accurate treatment of cerebrovascular malformations. Our overall objectives in this project are to (i) develop a novel approach based on deep neural networks that can classify and reconstruct 3D dynamic cerebral vasculature from 2D Digital Subtraction Angiography (DSA) image series, (ii) compose an AR visualization that will enhance the surgical view of the brain, and (iii) validate and evaluate our technology in real clinical settings. The rationale for this project is that such technology will provide a clear and interpretable visualization tool to surgeons that will support their decision-making process and reduce the time and complex spatial reasoning required to treat cerebrovascular malformations. To attain the overall objectives, the following three specific aims will be pursued: 1) develop and validate a method to classify artery and veins in DSA image series to visually disentangle AVMs, 2) develop and validate a method to build dynamic, virtual 3D model of cerebral vasculature from pairs of DSA image series and 3) build an AR visualization that aligns DSA image series with the surgical view and assess its impact providing surgical guidance. In addition, we will examine, through a clinical retrospective study, and through tests in the operating room on phantom data, the impact of this visualization in providing surgeons with guidance during cerebrovascular surgery. The proposed project is innovative because it will be possible to merge the true DSA-derived 3D cerebral hemodynamics with images of the brain surface seen through a surgical microscope. The proposed project is significant because it will provide visual guidance and confirmation to surgeons that will facilitate decision-making in the surgical treatment of complex AVMs. The results are expected to have an important positive impact because they will provide novel neuronavigation tools to improve the surgical treatment of cerebrovascular malformations and ultimately reduce the risks of intraoperative hemorrhaging and postsurgical deficits. Furthermore, the methods described here are cost-effective, adapted to low-resources settings, and can be easily implemented on a large scale, bringing advanced imaging techniques to far more patients.

NIH Research Projects · FY 2026 · 2024-03

ABSTRACT The goal of this project is to test the hypothesis that a cerebellar-striatal circuit regulates satiety in humans. Obesity currently affects 42% of U.S. adults, with projections that by 2030, 50% of U.S. adults will have obesity. These trends forecast a parallel increase in risk of obesity-related morbidity and mortality. Behavior, emergent from brain function, likely governs several primary sources of obesity, yet therapeutic intervention based on known brain circuits has not yielded dramatic success, suggesting a need to identify and rigorously evaluate neural circuits associated with appetite and food intake. We used a reverse-translational approach to identify the cerebellum as a regulator of appetite and feeding behavior. Animal studies determined that this effect is due to cerebellar regulation of food related reward activity in ventral striatum (VS). Our prior studies of this circuit in humans have been observational. Rigorous examination of the causal role of this circuit in modulating reward- related food intake in humans requires well-designed acute, mechanistic, state-of-the-art neuromodulation studies. Our group has evidence that cerebellar transcranial magnetic stimulation (TMS) can selectively modulate network function in cerebellar networks in healthy adults, and when applied therapeutically, can reduce symptom severity in patient populations. Thus, given the nascent nature of human data collected to date, we will integrate TMS and neuroimaging techniques to validate the role of the cerebellar-VS circuit in satiation in humans. We hypothesize that TMS modulation of the cerebellar-VS circuit will elicit neural and behavioral responses that are consistent with state-dependent cerebellar-induced satiation. These include increases in cerebellar activation, reductions in VS activation, and reduced food reward behaviors and intake. This proposal will innovatively combine cerebellar TMS with neuroimaging and behavioral measures of food-related reward under varying appetitive states. We will study the acute impact of cerebellar TMS on the satiation response to palatable food in neural systems and behavioral endpoints using a double-blinded, randomized, sham-controlled, mechanistic parallel trial involving cerebellar TMS neuromodulation in 150 healthy adults with BMIs spanning the healthy weight to obese range. The approach, completed under fasted and fed states to identify state-specific functioning of the cerebellar-VS circuit, will include functional MRI to ascertain neural reactivity to high-palatable food in the cerebellum and VS, as measures of target engagement. Food reward behavior will be measured using objective assessments and subjective ratings, in addition to ad libitum intake of palatable food. This proposal has public health impact because it will provide a mechanistic understanding of how this novel cerebellar-ventral striatal circuit functions to induce a reward-related satiation response in humans, which in turn will allow development of obesity therapeutics that target these validated and novel neurobiological pathways.

NIH Research Projects · FY 2025 · 2024-03

Project Summary Fibrous dysplasia (FD) affects approximately 1 in 5000 individuals and causes focal bone lesions that result in pain, deformity and fracture susceptibility. FD lesions are characterized by an accumulation of immature osteoblast lineage cells as well as robust osteoclastogenesis with increased bone turnover. Receptor activator of NFkB ligand (RANKL), the key cytokine driving osteoclast differentiation, is highly expressed by osteoblastic cells in FD lesions. Neutralizing antibody against RANKL, aRANKL, prevents osteoclast formation and decreases lesion size in mouse models of FD. In patients with FD, denosumab, which targets RANKL, is the only effective treatment. However, denosumab can inhibit skeletal growth and withdrawal can cause rebound bone resorption and fractures. Thus, alternative therapies are needed. If we could understand how aRANKL inhibits FD bone lesions, we might be able to discover new treatment targets. To investigate the effect of aRANKL antibody treatment in FD, we used an established mouse model where GnasR201H expression is induced post-zygotically in skeletal stem cells; bone lesions in this model are osteoblastic precursors which are a mosaic of wildtype and lineage traced mutant osteoblastic cells. We demonstrated that aRANKL inhibits the fibrotic bone lesions of FD in this model. At the cellular level, we demonstrated that aRANKL treatment altered the phenotype of Gnas mutant cells and also had a significant impact on wildtype osteoblastic cells, inhibiting proliferation and promoting differentiation of both. The cellular mechanism responsible for the effect of aRANKL is not known. While RANKL is best known for its role in driving osteoclast differentiation, it has many other biologic functions. For example, RANKL is critical for the development of lymph nodes and some specialized epithelial cells, and it can stimulate osteoblast differentiation through reverse signaling. Thus, we hypothesize that RANKL may promote FD bone lesion pathogenesis in an osteoclast-independent manner and that the critical target of aRANKL is RANKL itself. We will test this hypothesis by comparing the efficacy of osteoclast inhibition with anti-CSF1R (which blocks a key survival factor for osteoclasts) with aRANKL in inhibiting FD lesions in this model. The molecular mechanism by which aRANKL treatment alters the cellular phenotype of Gnas mutant cells and wild-type cells in FD lesions is unknown. To investigate the pathways downstream of aRANKL treatment, we propose to determine the transcriptional changes induced in Gnas mutant cells and adjacent wild- type osteoblastic cells using a spatial transcriptomic approach using the GeoMx digital spatial profiler platform. Through a combination of determining whether aRANKL efficacy is independent of osteoclast inhibition and molecular investigation of tissue and cell level response to aRANKL, this proposal will significantly advance our understanding of FD pathogenesis and identify additional pathways that could be targeted for treatment.

NIH Research Projects · FY 2026 · 2024-03

Project Summary/Abstract The proposed project is focused on the mechanism controlling posterior elongation of the vertebrate embryo, a process which is still poorly understood. In amniotes such as birds or mammals, the embryonic body forms sequentially in a head-to-tail sequence. Progressively more posterior territories are laid in the wake of regression movements of the primitive streak and tail bud (TB) [1]. Unlike anterior regions which mostly depend on convergence extension, posterior elongation of the embryonic body of amniotes relies on volumetric growth. Our past work showed how the paraxial mesoderm, which forms the skeletal muscles and vertebrae, drives these posterior elongation movements [2]. We demonstrated that a posterior-to-anterior gradient of FGF signal- ing established in the most posterior region of the paraxial mesoderm (called presomitic mesoderm (PSM)) im- poses a parallel gradient of random cell motility in this tissue. This gradient is necessary for posterior elongation of the embryo. Our findings led us to propose that the PSM can generate posteriorly oriented compression forces that trigger axial elongation in response to the FGF gradient [3]. Indeed, we demonstrated that an isolated PSM can generate FGF-dependent elongation forces when confined in a PDMS microchannel [4]. Our preliminary data demonstrated that the extracellular volume (EV) is increased in the posterior PSM compared to the anterior. We showed that hyaluronic acid (HA), a naturally occurring polymer composed of repeating disaccharides, is an important component of the PSM extracellular matrix required for PSM elongation [4]. Together, our results sug- gest a model whereby FGF promotes cell motility, triggering an HA-dependent increase in EV, generating com- pression forces extending the embryo posteriorly. Here, we will investigate the role of HA in embryonic elongation. We will combine in vivo experiments in the chicken embryo with in vitro studies in novel human iPS-derived PSM organoids that we recently established [5]. We will first analyze how HA signals to PSM cells to control their motility in vivo in the chicken embryo. We will then examine whether this plays a role in the control of the EV. We will also leverage our novel system of human iPS-derived PSM organoids (called segmentoids), which form elongated structures recapitulating PSM development [5]. This system offers the possibility to genetically manipulate iPS cells and provides the unprec- edented opportunity to explore the role of HA in the control of PSM elongation. Finally, we will use a comple- mentary 3D model of human iPS-derived somites (called somitoids) to probe HA’s role in PSM epithelialization and somite formation. Our studies will shed light on the process of axis elongation and elucidate the role of HA, a poorly studied ubiquitous glycosaminoglycan, in this morphogenetic process. This work will be informative about human malformations associated to defective axis formation, such as caudal agenesis or spina bifida [6].

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY/ABSTRACT This NIH K23 proposal details a comprehensive five-year training plan for mentored patient-oriented research career development and research to address a major problem in neuromuscular medicine. Over half of patients with clinically suspected Mendelian myopathies do not have a molecular diagnosis, largely due to two challenges: detecting pathogenic non-coding variants, and resolving the pathogenicity of ultra-rare missense variants. Here I propose to address this challenge through the application of genome sequencing (Aim 1), transcriptome sequencing (Aim 2), and proteomic (Aim 3) methods to evaluate the strengths and weakness of each method and to improve the diagnostic yield from a cohort of approximately 200 individuals with unsolved Mendelian myopathies. I hypothesize that since the genetic architecture of Mendelian myopathies implicates large genes that genome sequencing combined with RNA-sequencing (RNA-seq) and proteomics can mitigate the current low diagnostic yield after clinical evaluation of Mendelian myopathies. Through these approaches, I will define best practices in applying technologies in the diagnostic evaluation, discover novel disease variants and genes, and expand our understanding of the genetic architecture of these heterogeneous disorders. Gaining the analytical skills to evaluate the real-world application of multiomic methods will complement both my prior expertise in gene discovery for neurologic disorders and my clinical training in neuromuscular medicine. I am uniquely positioned within the collaborative environment between Brigham and Women’s Hospital, Boston Children’s Hospital, Broad Institute, and Harvard Medical School to facilitate my transition to an independent physician-scientist with a long-term translational research goal of developing a center of excellence in the clinical characterization, molecular diagnosis, and evaluation of targeted gene therapy candidates for patients with Mendelian myopathies.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY While opioid drugs are very effective for controlling pain, they are highly addictive. The current clinical decision making for post-surgical opioid prescribing is still based on oversimplified information. Both opioid use disorder and opioid-related pain response have strong genetic underpinnings. A better system is needed to guide the clinical use of genotype data and facilitate better-informed opioid prescribing decision-making. Electronic health record (EHR) offer a largely untapped source of information to conduct clinically oriented studies. By integrating genomic data and EHR dataset in large-scale clinical Biobank, we can now perform "clinical-driven genomic research". This approach is particularly robust for complex diseases with a genetic susceptibility. I propose to develop a model to recommend optimal opioid dosage (oral morphine milligram equivalents) for post-surgical pain relief and predict the risk of post-discharge harmful outcomes. The large-scale clinical and genomic databases from the Mass General Brigham (MGB) healthcare system, including eight hospitals that share a centralized database, will be utilized for model development. The proposed research will not only have strong immediate potential to improve clinical practice in this urgent area but will also provide strong preliminary work to support my first NIH R01 application. From the previous projects, I have developed opioid use disorder clinical phenotypes and identified associated genetic markers. I have also developed machine learning models to predict risks of complex diseases and explored clinical application potentials. The proposed project will combine my existing analytical skills with the newer expertise that I seek to develop through this award program, including a nuanced understanding of analgesic strategies, addiction, and implementation of clinical decision support systems, as well as artificial intelligence applications in healthcare system. I have invited several established investigators to form a strong, multi-disciplinary mentorship team. They include experts in the fields of addiction, pain management, genetic analysis, implementation methods and artificial intelligence techniques. With their support, I intend to dedicate my longer-term research career to study functional genomics in the fields of opioid addiction and pain by using integrated analysis of harmonized big data. The K01 program outlined in this application will provide timely and strategic support for my continued advancement towards those goals.

NIH Research Projects · FY 2026 · 2024-02

Oxidative stress is a hallmark of neurodegeneration and has been implicated in the pathobiology of vascular contributions to cognitive impairment and dementia (VCID). Our “chemogenetic” approach allows us to dynamically modulate reactive oxygen species (ROS) in target cells in vivo using the yeast enzyme D-amino acid oxidase (DAAO) to produce the ROS hydrogen peroxide (H2O2). We generated two new transgenic mouse lines that express DAAO by crossing a conditionally activatable DAAO transgenic mouse line (which we developed) with commercially available mouse lines expressing Cre recombinase under control of two distinct putatively “endothelial cell-specific” Cdh5 or Tie2 promoters. Both the DAAO-TGCdh5 and DAAO-TGTie2 lines express DAAO and generate H2O2 in endothelial cells. Within 2 days of providing D-alanine to DAAO-TGCdh5 mice, the animals develop a striking sensory ataxia, and have a highly specific pattern of neurodegeneration and mitochondrial disarray in dorsal root ganglia and nodose (vagal sensory) ganglia. Importantly, DAAO-TGCdh5 mice treated with D-alanine also develop cardiac hypertrophy. The combination of sensory neuropathy and cardiac hypertrophy is similar to the phenotype of Friedreich’s ataxia, the most common form of hereditary ataxia in humans. By contrast, the DAAO-TGTie2 transgenic line, which expresses DAAO in endothelium under control of the Tie2 promoter, shows no ataxia, has no transgene expression in DRG, and does not develop cardiac hypertrophy. But the DAAO-TGTie2 mice does develop marked disruption of the blood barrier following long-term D-alanine feeding. Here we propose studies to test the hypotheses that neurovascular oxidative stress leads to neurodegeneration, BBB disruption, cognitive dysfunction, and cardiac hypertrophy. We propose to pursue transcriptomic, proteomic, metabolomic, and biochemical studies of DRG, nodose ganglia, brain, and cardiac tissues following chemogenetic oxidative stress in vivo, and we will study the behavioral and physiological phenotypes in the DAAO-TGCdh5 and DAAO-TGTie2 mouse lines. We will identify the temporal sequence and the molecular mechanisms whereby neurovascular oxidative stress causes degeneration of DRG and nodose ganglia and leads to cardiac hypertrophy. We will establish the pathways by which vascular oxidative stress disrupts the BBB and leads to cognitive impairment. The Specific Aims are: Aim 1: Identify the molecular mechanisms whereby neurovascular oxidative stress causes degeneration of DRG; Aim 2: Characterize the molecular processes whereby neurovascular oxidative stress causes disruption of the blood-brain barrier and leads to cognitive impairment; Aim 3: Define the pathways whereby neurovascular oxidative stress causes cardiac hypertrophy. The proposed studies may lead to the identification of new pharmacological targets for prevention and treatment of Alzheimer’s Disease and related dementias, Friedreich’s ataxia, adverse cardiac remodeling, and the many other chronic disease states in which oxidative stress has been implicated.

NIH Research Projects · FY 2025 · 2024-02

PROJECT SUMMARY The overall goal of this proposal is to systematically characterize the cellular functions of transcription factor isoforms. Transcription factors (TFs) are master regulators of gene expression and as such play key roles in a variety of biological processes, including cell growth and differentiation. The human genome is estimated to harbor ~1600 TF genes; however, most of these are expressed as a series of protein isoforms arising from mRNAs with alternative starts, ends, or splicing. Though a handful of alternative TF isoforms are known to play functionally important (and distinct) roles in the cell, the overwhelming majority— thousands of proteins—remain entirely uncharacterized. Moreover, splicing aberrations are a hallmark of cancer, and mis-expression of TF isoforms can contribute to tumorigenesis. Thus, decoding the roles of TF isoforms is key to a systems-level understanding of gene regulatory networks (GRNs) in development and disease. In my previous work, I found widespread changes in DNA binding, protein-protein interactions, and transcriptional activation in exogenous assays across a collection of >700 TF isoforms. These results underscore the need to perturb and characterize TF isoforms in their endogenous cellular context to truly understand their roles in GRNs. Functional genomics approaches such as high throughput perturbation screens have revolutionized our understanding of gene functions. I aim to apply and extend these functional genomics approaches to study isoforms, which have remained elusive due to technical limitations. I propose to use a combination of state-of-the-art genomic technologies to decode the functions of TF isoforms in breast cancer cells. I will leverage long read RNA-sequencing to perform rigorous isoform-aware analyses (Aim 1) and isoform-specific high throughput experiments (Aims 2 and 3). In Aim 1, I will re-analyze existing CRISPR/Cas9 knock-out databases from the Cancer Dependency Map Consortium to identify candidate isoform-specific phenotypes in breast cancer. In Aim 2, I will establish a platform for robust and specific knock-down of individual isoforms in mammalian cells using RNA-targeting CRISPR/Cas systems. In Aim 3, I will use tunable libraries of TF ORFs to quantitatively tune isoform expression across a range of physiological levels. The proposed work includes technology development in the mentored K99 phase, which I will then leverage during the independent R00 phase to probe the mechanisms of TF isoform function in breast cancer via single-cell screening approaches. Successful execution of these aims will begin to decode the “dark matter” of alternative TF isoforms, laying a foundation for future studies of alternative isoforms more broadly. By combining new training in high- throughput screening approaches during the K99 phase with my existing expertise in gene regulation, TF biology, and bioinformatics, this transition to independence award will position me well to start my own research group that uses interdisciplinary genomic methods to probe GRNs in development and disease.

NIH Research Projects · FY 2026 · 2024-02

Project Summary/Abstract Current inflammatory bowel disease (IBD) therapeutic approaches are insufficient to maintain long-term immune homeostasis and effectively recalibrate T helper cell imbalances in patients. There remains an unmet clinical need for new strategies that sustain immune tolerance in IBD. The programmed death 1 (PD1) pathway has emerged as a critical inhibitory signal which controls T cell responses and maintains immune homeostasis. Altered PD1 signaling can predispose mice and humans to autoimmunity. For example, PD1 blockade can cause colitis in mice and humans. Recently, we identified that Smad7, a major molecule implicated in IBD, sustains intestinal inflammation in mice by limiting PD1 signaling, thereby dampening PD1-induced Tregs. Given the critical role of PD1 in limiting tissue inflammation, PD1 represents a therapeutic target of high clinical interest. We found that enhancing PD1 signaling via recombinant human PDL1-Fc or PDL2-Fc (PD1 agonist) promotes de novo human Treg induction and stability. Interestingly, we also found that agonizing PD1 in myeloid cells inhibits inflammatory cytokines that are known to promote Th1 and Th17 development and destabilize Tregs during IBD. In our effort to identify factors that upregulate PD1, we found that IL-2 directly induces PD1 on human T cells. Excitingly, we found that a combination of low-dose IL-2 with PD1 agonist synergistically promotes human Tregs. Based on these findings, we will test our hypothesis that PD1 agonist monotherapy could effectively restore immune tolerance by directly enhancing Treg homeostasis while quenching inflammatory T cell and myeloid cell responses. We also hypothesize that combining low-dose IL-2, which induces PD1 on T cells, with PDL1/2-Fc will synergistically boost Treg responses to better treat IBD. Because IL-2 can expand inflammatory T cell and myeloid cell responses, combining PD1 agonist with low-dose IL-2 may also restrain this undesired IL-2-induced inflammation. This is particularly exciting, since low-dose IL-2 for Treg induction has shown some efficacy in currently advancing clinical trials in IBD and other contexts, and strategies to improve low-dose IL 2 therapy are being actively pursued. In Aim 1, we will test the translational relevance of PD1 agonist monotherapy and combination therapy with low-dose IL-2 by treating IBD patient T cells in vitro and T helper cell-specific PD1- deficient (KO) mice with colitis. In Aim 2, we will test the translational relevance of PD1 agonist monotherapy and combination therapy with low-dose IL-2 by treating IBD patient myeloid cells in vitro and myeloid cell-specific PD1-deficient (KO) mice with colitis. In Aim 3, we will test the translational relevance of PD1 agonist monotherapy and combination therapy with low-dose IL-2 by treating translationally relevant humanized mice with colitis. In summary, we will explore the efficacy of a never tested PD1 agonist/low-dose IL-2 combination therapy strategy in IBD to address unanswered questions around how PD1 agonism promotes human immune tolerance, the translational potential of PD1 agonist therapeutic strategies, and how to implement them in IBD.

NIH Research Projects · FY 2026 · 2024-02

Project Summary The long-term objective of this project is to develop Dr. Romanos-Nanclares’s capacity to conduct studies that integrate dietary and lifestyle factors with multi-omics data and traditional biomarkers to better understand the biological pathways underlying the etiology of aggressive forms of breast cancer and to advance the field of precision nutrition for cancer prevention. The proposed research combines ‘omics methodologies with advanced statistical techniques to investigate the role of chronic inflammation in the etiology of breast cancer. Chronic inflammation has been identified as a possible risk factor in the development of breast cancer, including estrogen receptor negative breast cancer, which is a subtype of breast cancer that is associated with poorer prognosis and is less responsive to hormonal therapies. The foundation for this proposal is based on our recent findings where a higher adherence to an Empirical Dietary Inflammatory Pattern (EDIP) score (or a more pro- inflammatory diet) was associated with increased breast cancer risk, particularly for tumors lacking estrogen receptor expression. Leveraging the unique design and wealth of resources in the Nurses’ Health Study (NHS) and Nurses’ Health Study II (NHSII), with repeated dietary measures and prediagnostic plasma specimens, this application will combine dietary and ‘omics data from multiple biological dimensions with cutting-edge epidemiological and bioinformatics methodologies to examine the relationship between chronic inflammation and invasive breast cancer risk and estrogen receptor subtypes. In Aim 1 (K99), she will examine the associations of plasma metabolomic profiles of EDIP with subsequent risk of breast cancer in the NHS and NHSII among 1,997 cases and their matched controls. In Aim 2 (R00), she will use an agnostic approach to identify the associations of plasma proteomics with risk of breast cancer in the NHS and NHSII among 200 cases and their matched controls. In Aim 3 (R00), she will integrate multi-omics data, including metabolomics, proteomics, and transcriptomics, to characterize the role of inflammation in breast cancer etiology. This research will provide novel evidence, enhance our understanding of the biological mechanisms involved in breast cancer etiology, and help identify new prevention strategies for more aggressive forms of the disease. Dr. Romanos-Nanclares’s scientific aims are supported by three training objectives that will allow her to advance her trajectory toward becoming an independent investigator: 1) single-omics and multi-omics methodologies and advanced analytics; 2) proteomics study design and execution; and 3) breast cancer etiology and pathophysiology. The training environment at Brigham and Women’s Hospital will provide exceptional support for Dr. Romanos-Nanclares as she pursues these development and scientific goals. She has assembled an outstanding interdisciplinary mentoring team with expertise in cancer epidemiology, multi-omics bioinformatics, breast cancer pathology, proteomics, and metabolomics. This K99/R00 award will help Dr. Romanos-Nanclares acquire the knowledge and experience necessary to launch her career as an independent investigator.

NIH Research Projects · FY 2025 · 2024-02

Project Summary The long-term objective of this project is to develop Dr. Romanos-Nanclares’s capacity to conduct studies that integrate dietary and lifestyle factors with multi-omics data and traditional biomarkers to better understand the biological pathways underlying the etiology of aggressive forms of breast cancer and to advance the field of precision nutrition for cancer prevention. The proposed research combines ‘omics methodologies with advanced statistical techniques to investigate the role of chronic inflammation in the etiology of breast cancer. Chronic inflammation has been identified as a possible risk factor in the development of breast cancer, including estrogen receptor negative breast cancer, which is a subtype of breast cancer that is associated with poorer prognosis and is less responsive to hormonal therapies. The foundation for this proposal is based on our recent findings where a higher adherence to an Empirical Dietary Inflammatory Pattern (EDIP) score (or a more pro- inflammatory diet) was associated with increased breast cancer risk, particularly for tumors lacking estrogen receptor expression. Leveraging the unique design and wealth of resources in the Nurses’ Health Study (NHS) and Nurses’ Health Study II (NHSII), with repeated dietary measures and prediagnostic plasma specimens, this application will combine dietary and ‘omics data from multiple biological dimensions with cutting-edge epidemiological and bioinformatics methodologies to examine the relationship between chronic inflammation and invasive breast cancer risk and estrogen receptor subtypes. In Aim 1 (K99), she will examine the associations of plasma metabolomic profiles of EDIP with subsequent risk of breast cancer in the NHS and NHSII among 1,997 cases and their matched controls. In Aim 2 (R00), she will use an agnostic approach to identify the associations of plasma proteomics with risk of breast cancer in the NHS and NHSII among 200 cases and their matched controls. In Aim 3 (R00), she will integrate multi-omics data, including metabolomics, proteomics, and transcriptomics, to characterize the role of inflammation in breast cancer etiology. This research will provide novel evidence, enhance our understanding of the biological mechanisms involved in breast cancer etiology, and help identify new prevention strategies for more aggressive forms of the disease. Dr. Romanos-Nanclares’s scientific aims are supported by three training objectives that will allow her to advance her trajectory toward becoming an independent investigator: 1) single-omics and multi-omics methodologies and advanced analytics; 2) proteomics study design and execution; and 3) breast cancer etiology and pathophysiology. The training environment at Brigham and Women’s Hospital will provide exceptional support for Dr. Romanos-Nanclares as she pursues these development and scientific goals. She has assembled an outstanding interdisciplinary mentoring team with expertise in cancer epidemiology, multi-omics bioinformatics, breast cancer pathology, proteomics, and metabolomics. This K99/R00 award will help Dr. Romanos-Nanclares acquire the knowledge and experience necessary to launch her career as an independent investigator.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY/ABSTRACT Skull base and face imaging accounts for nearly 20% of clinical neuroradiology examinations. But clinical MRI of the skull base and face continues to rely primarily on conventional 2D T1-weighted and T2-weighted pulse sequences and fat suppression techniques. These techniques generate suboptimal tissue contrasts with suboptimal spatial resolution. The long acquisition times often lead to motion artifact. Contrast weightings and rapid acquisitions tailored to the uniquely small structures of the skull base and face would expand the clinical utility of MRI for diagnosing pathologies of these regions. Dr. Guenette’s goal for this four-year K08 mentored career development award is to gain the skills needed to develop comprehensive skull base- and face-specific MRI sequences and protocols that optimize clinically relevant contrast-to-noise and simultaneously acquire relaxometry biomarkers that are co-registered to the anatomic images at the voxel level with markedly reduced scan times compared with current standard-of-care skull base and face MR imaging. Dr. Guenette is a neuroradiologist with skull base and face sub-specialization at Brigham and Women’s Hospital. His research has included optimizing product pulse sequences for imaging skull base and face structures. The training objectives and research activities of this proposal will provide Dr. Guenette with the mentorship, time, and resources to learn and apply MRI physics, pulse sequence design, and image processing. The main research expertise of Dr. Guenette’s mentor, Dr. Bruno Madore, an MRI physicist, lies in the development of novel acquisition and image reconstruction strategies for MRI. The co-mentor, Dr. Ravi Uppaluri, is an otolaryngology-head/neck surgeon physician-scientist with complementary clinical expertise. In Aim 1 Dr. Guenette will: build a normative atlas of quantitative T2, T1, and relative proton density values of anatomic structures in the skull base and face; use this atlas to generate and test theoretically optimal contrasts for nerve imaging; establish normative T2, T1, and relative proton density values of head and neck squamous cell carcinoma (HNSCC) tumors to generate and test optimal contrasts for visualizing and segmenting HNSCC tumors; and test the biomarker potential of T2, T1, and relative proton density measures in HNSCC. In Aim 2 he will: develop a novel pulse sequence that incorporates scanner gradient strength as a parameter to further decrease scan times and evaluate synthetic pulse sequences across 1.5T, 3T, and 7T field strengths. The training and data will facilitate Dr. Guenette’s long-term goal to independently lead an NIH-funded laboratory focused on developing and implementing more rapid qualitative MR anatomic imaging and quantitative MR methods specifically tailored to the skull base, face, and neck structures and pathologies.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY/ABSTRACT: Recent FDA accelerated approvals of two anti-amyloid antibody treatments, aducanumab and lecanemab, for early-stage Alzheimer’s disease (AD) provide the first disease-modifying treatments to date, albeit with moderate slowing of cognitive decline, and vascular side effects known as Amyloid Related Imaging Abnormalities due to edema (ARIA-E) or microhemorrhages (ARIA-H) in ~12-35% of patients, especially ApoE4 carriers. While the cause of ARIA is unknown, it has been suggested that Ab clearance by anti-amyloid antibodies is mediated by perivascular drainage which may transiently lead to amyloid accumulating in the blood vessel wall and inducing inflammation, which in turn may increase BBB breakdown, leading to edema or microhemorrhages (Cogswell et al., 2022). Therefore, there is still a large unmet need to improve cognitive efficacy and safety of anti-amyloid immunotherapy. Semaglutide is a Glucagon-Like Peptide-1 Receptor (GLP-1R) agonist, that is FDA approved for the treatment of Type 2 Diabetes Mellitus and obesity, that has strong anti-inflammatory, neuroprotective and pro-vascular health effects. Previous studies have shown beneficial effects of semaglutide in stroke models, diabetes models, LPS models and other disease models. Novo Nordisk, the inventor of semaglutide, is conducting two large Phase 3a clinical trials to test the efficacy of semaglutide in early-stage AD (EVOKE, EVOKE+). While other GLP-1R agonists have shown benefits in AD mouse models, semaglutide, a longer lasting form, has not yet been tested in amyloid AD mouse models. The goal of this project is to determine whether the combination of semaglutide with anti-amyloid antibody therapy will enhance the efficacy beyond that of either treatment alone and attenuate the vascular side effects seen with anti-amyloid antibodies. We propose the following 3 Aims: 1. We hypothesize that semaglutide alone will reduce or reverse inflammation and neurodegeneration and spare cognition in 2 amyloid AD-like mouse models: 5XE4 mice which overexpress human mutant APP/PS1 on a human ApoE4/4 background and APPSAA knockin mice which express physiologic levels of human mutant APP. Mice will be dosed for 12 weeks at 2 ages – pre-plaque and after robust plaque and vascular amyloid accumulation. 2. We hypothesize that combination therapy with semaglutide and anti- amyloid antibodies (murine versions of lecanemab and donanemab) will enhance efficacy compared to either treatment alone in the 5XE4 model which develops abundant plaques, vascular amyloid, inflammation and cognitive decline with aging. 3. We hypothesize that acute semaglutide co-treatment with a murine precursor to bapineuzumab (3D6) shown to cause a high amount of ARIA, will mitigate microhemorrhagic side effects in aged 5XE4 mice. Outcomes include behavioral testing, pathological and biochemical measures, bulk RNAseq and in- depth analysis of transcriptomic changes within cell populations in cortex and hippocampus, and mapping of the mouse data to human data available on the NIH-sponsored AMP-AD portal. If successful, combination trials in humans could be initiated to improve the efficacy and safety of anti-amyloid antibody immunotherapy.

NIH Research Projects · FY 2025 · 2024-01

Congenital cytomegalovirus (cCMV) infection is the most common congenital infection in the US, affecting one in every 200 infants each year. Approximately 10% of newborns impacted by cCMV have symptoms at birth, such as small size for gestational age, jaundice, petechiae/purpira, and hepatosplenomegaly. Symptomatic cCMV is associated with a higher risk of long-term sequelae and mortality. Despite its significant clinical consequences, cCMV remains one of the least recognized and understood viruses that can spread from pregnant patients to their unborn children. There is an urgent need for high-quality evidence on the risks of long-term neurologic and non-neurologic sequelae and on the benefits of available treatments for these outcomes among cCMV infected infants. While neurologic sequelae are commonly reported in cCMV infected infants, the available evidence has significant limitations (e.g., limited follow-up and sample sizes) preventing accurate quantification of the risks of long-term neurologic sequelae attributable to cCMV infection. While several studies have reported cases of non- neurological adverse outcomes among infants and children with cCMV, including hepatitis, pneumonitis, and myocarditis, it is currently unknown whether cCMV infection leads to long-term non-neurologic sequelae. Whereas randomized controlled trials have demonstrated the effectiveness of ganciclovir and valganciclovir in preventing hearing deterioration, improving hearing and early neurodevelopmental milestones in symptomatic infants before the age of 2, it remains unclear whether antivirals also reduce the risks of late-onset hearing loss and long-term neurologic and non-neurologic sequelae. Using nationwide cohorts of publicly (Medicaid, 2000-2020) and privately (MarketScan, 2003-2021) insured neonates (≈ 29 million, both cohorts to be updated as new data become available during the course of the study), which provide a unique, highly representative sample of the overall US neonatal and pediatric population, as well as sophisticated epidemiologic design and analytic approaches and novel tree-based scanning signal detection methods, we will address the following specific aims: (i) to quantify the risks of neurologic long-term sequelae among infants with clinically recognized cCMV; (ii) to conduct a systematic surveillance of newborns with clinically recognized cCMV to detect signals of adverse long-term non-neurologic health consequences. For both aims, the effect of antiviral treatment on the risk of these neurologic and non- neurologic outcomes will be evaluated. By harnessing the power of existing real-world data, the proposed studies will have an immediate and important public health impact by informing the value of universal screening, early antiviral treatments and vaccines, as well as by raising awareness among patients and clinicians.