Univ Of Massachusetts Med Sch Worcester

NIH Research Projects · FY 2026 · 2026-02

Abstract. The Bacillus Calmette-Guerin (BCG) vaccine has been in use for over a century and is currently administered in more than 200 countries to protect against tuberculosis (TB). Despite this, TB remains one of the leading causes of death due to an infectious disease. Efforts to improve or enhance BCG has been met with limited success in part due to the lack of an immune correlate that is associated with vaccine-induced protection in the human population. Although it is unclear why BCG remains ineffective against adult pulmonary TB, traditional animal models such as the C57BL/6 (B6) mice display a relatively homogenous Th1 response following BCG vaccination and fail to replicate the heterogeneous clinical response observed in human populations. Using the Collaborative Cross (CC) mice, we have demonstrated that host genetics is a critical determinant of both primary susceptibility and BCG-induced protection against Mtb, with the hope of using the CC model to identify immune signatures associated with protection that is not observed in the traditional B6 mice. Within several CC strains, we have identified a hybrid-Th1/17 population that is correlated with BCG-induced protection against Mtb, suggesting that this is a signal that is associated with vaccine- induced protection across multiple host genotypes. Using CC037, which is the strain that is the most protected against Mtb following BCG vaccination and has the strongest hybrid-Th1/17 signal, we will ask how BCG vaccination modifies host immunity in CC037 to generate this population in response to Mtb infection. As dendritic cells (DC) are considered the primary cell type that bridges the innate and adaptive immune systems, our overarching hypothesis is that modification of the DC compartment by BCG is leading to the generation of hybrid-Th1/17 response in CC037 mice. We will ask whether the ability to generate a hybrid-Th1/17 response is inherent to the host T cells or is due to certain signals produced by DC in response to BCG vaccination. We will also use a combination of scRNA-Seq and ATAC-Seq to examine how BCG modifies host T cell and DC responses to Mtb in CC037, from both a transcriptional and epigenetic approach. CC037 represents an important tool for understanding how hybrid-Th1/17 responses are generated and how they contribute to protective immunity against Mtb. This study aligns with the NIH NOT-AI-24-054, where we expect our study to identify novel mechanisms that will lead to the induction of this response and enhance TB vaccine development in the context of diverse genetic backgrounds.

NIH Research Projects · FY 2026 · 2026-02

Project Summary/Abstract The world's population is aging rapidly, and by 2050, the population over 60 years will exceed 2 billion people or 25% of the total global population. Tissue repair is critical for the survival of all organisms. Aging causes a gradual decline in tissue integrity, partly due to a decline in stem cell function, a major hallmark of aging. Due to its accessibility and temporal predictability, cutaneous wound healing provides a valuable model framework to characterize the effects of aging on tissue injury1. Aging disrupts the precisely coordinated immune cell response that orchestrates the three phases of cutaneous wound healing. However, it is unknown whether this disruption is initiated by the aging of the tissue itself or the aging of the immune system. Recently, a breakthrough study showed that the aging immune system or “immunosenescence” precedes and drives organ aging2. Thus, an inference from this concept is that the mechanism by which aging impairs wound healing is largely dependent on how aging impairs the immune system. Our long-term goal is to identify how aging impairs immune cell function and show that we can restore youthful wound healing by reversing immune aging. Hematopoietic stem cells (HSCs) are remarkable cells in our bone marrow that make at least one trillion new blood cells daily. We have shown that HSCs produce all of our circulating immune cells and regulate their gene expression by “epigenetic reprogramming”. We will use sophisticated “single-cell epigenomics,” some of which were developed in our laboratories, to characterize how aging affects the gene expression of individual immune cells and how that expression is regulated by epigenetic modifications. We have three Aims: Aim 1: Determine if aging impairs wound healing by an HSC-Autonomous mechanism. Aim 2: Identify the aging-specific- HSC oxidant stress that drives immune aging and impairs wound healing. Aim 3: Identify the master epigenetic enzyme(s) in aged HSCs that epigenetically reprogram the gene expression of wound macrophages and their cross-talk with fibroblasts and keratinocytes. Once we identify the “master epigenetic enzyme” affected by aging in HSCs that reprograms the gene expression of immune cells, we will reverse the effects of aging on the master epigenetic enzyme to restore youthful wound healing. The fact that the effects are “epigenetic” implies that the effects of aging, at least on the immune system, are reversible and, by proof of principle, would open the door to new molecular therapies to reverse the effects of aging.

NIH Research Projects · FY 2026 · 2026-01

Abstract Current therapeutic interventions for treating neurodegenerative disorders, including Alzheimer’s Disease (AD), frontotemporal dementia (FTD), multiple sclerosis (MS), and Amyotrophic lateral sclerosis (ALS), are highly limited in both number and efficacy, suggesting novel approaches are needed. Most prior research has focused on developing strategies for treating each of these diseases individually, but these endeavors have largely failed in advancing efficacious therapeutics. We have recently generated exciting data that offers the promise to overcome this gap in neurodegenerative research. We find that genetic deletion of one of the members of the membrane spanning 4a (Ms4a) family of genes, Ms4a6c, significantly improves cognitive, behavioral, and cellular phenotypes in mouse models of AD, FTD, ALS, and MS. Moreover, we and others have demonstrated that polymorphisms in Ms4a genes are strongly linked to altered susceptibility to neurodegenerative disorders, including ALS and AD. The link between Ms4a genes and neurodegeneration has been best studied in AD, where current data suggest that Ms4a polymorphisms are among the strongest genetic modifiers of AD risk. Together, this research suggests that therapeutic strategies targeting Ms4a genes are likely to be beneficial in treating many neurodegenerative disorders. However, to date we have only examined the effect of deleting a single Ms4a gene on neurodegeneration. Although Ms4a6c deletion results in significant improvement in all phenotypes examined across many neurodegenerative disorders, the phenotypic rescue remains incomplete. The Ms4a family consists of 17 genes, and other family members, including Ms4a4a, Ms4a4e, and Ms4a6e have also been strongly genetically linked to altered risk of developing neurodegenerative disorders. However, most of these AD associated polymorphisms localize to non-coding regions of this gene family, and their effect on MS4A function remains largely unknown, making it unclear whether loss of function or gain of function of these genes is protective against neurodegeneration. We have found that similarly to Ms4a6c, the expression of many other Ms4a genes is upregulated in multiple mouse models of neurodegeneration. Together, these observations have led us to hypothesize that deletion of other Ms4a family members might also improve neurodegenerative phenotypes, and we have generated significant pilot data in support of this hypothesis. Our first aim will therefore take advantage of novel mouse genetic reagents we have generated to enable us to probe the effect of deleting (or over-expressing) other Ms4a family members individually or deleting multiple Ms4a family members simultaneously on neurodegenerative phenotypes. In our second aim, we will build on considerable pilot data to elucidate the mechanisms by which Ms4a genes regulate neurodegeneration, a process which to date remains entirely unexplored. Together, these aims will provide significant insight into the role that Ms4a genes play across neurodegenerative diseases and take us one significant step closer to developing therapeutic approaches targeting these genes.

NIH Research Projects · FY 2026 · 2026-01

Project Summary The brain’s ability to process sensory cues and adapt behavior occurs in virtually all animals and is crucial to survival. Understanding how individual neurons integrate multimodal inputs to orchestrate complex behaviors remains largely unknown. The goal of my work is to investigate molecular mechanisms regulating sensory input to a single neuron in the nematode, Caenorhabditis elegans. The robust genetics, and invariant neural organization of C. elegans allows for single-neuron mechanistic investigation in intact, behaving animals. My project aims to uncover molecular regulators encoding foraging behavior. Upon immediate removal of food, animals engage in area-restricted search (ARS) behavior, characterized by increased turning events to maximize re-entry to food. Previous work in the Francis lab has demonstrated that the NLP-12 neuropeptide, released solely from the interneuron DVA, drives ARS. The lab has further suggested that dopaminergic signaling to the DVA couples to NLP-12 release, although a direct mechanism has not been elucidated. My preliminary data suggest that the TRPN (NOMPC) non-selective ion channel, TRP-4, potentially inhibits NLP- 12 release. This suggests that dopaminergic, and proprioceptive signaling via TRP-4, act antagonistically to modulate NLP-12 release and drive ARS. In Aim 1, I will investigate how dopaminergic signaling to the DVA drives DVA activity, NLP-12 release, and ARS. In Aim 2, I will uncover where TRP-4 regulates ARS, DVA activity, and NLP-12 release In Aim 3, I will investigate how disruptions in both dopaminergic and TRP-4 signaling regulate ARS, DVA activity and NLP-12 release. This project will define how sensory information encoding food availability and proprioceptive information are integrated to regulate neural activity, neuropeptide release, and behavior.

NIH Research Projects · FY 2025 · 2025-09

Project summary/abstract It is well-understood that life-relevant changes in independence occur as early warnings of lost cognitive resilience, and eventually Alzheimer's Disease and related dementias (ADRD). We and others demonstrated that medication self-administration (MSA) provides exactly the opportunity needed for early identification of ADRD. However, several knowledge gaps hinder routine assessment of this critical health self-management skill in current care. We lack studies directly comparing self-reported measures and objective, convenient MSA assessments in the general population. In addition, the impact of CVD and cognitive risk factors on MSA errors and MSA overestimation, strong predictors of memory performance and daily life functional independence, is unclear. To address these challenges, we propose to perform an objective MSA assessment in the Framingham Heart Study. This cohort has well-characterized cognitive assessment for up to three decades. An estimated 1185 surviving participants from the second-generation and Omni 1 Framingham Heart Study cohorts are expected to participate as part of their 11th /6th comprehensive health examination, starting September 2025. Our central hypothesis is that MSA errors and self- overestimation are early indicators of disabling brain and behavior changes. In Aim 1, we will cross-sectionally associate MSA errors and MSA self-overestimation, using the Hopkins Medication Schedule and a visual vertical scale, with behavioral- (neuropsychological performance) and brain-based ADRD biomarkers (atrophy, white matter change). In Aim 2, we will associate MSA assessment with a trajectory of cognitive decline on the Mini-Mental State (MMSE) and neuropsychological testing, as occurs in ADRD. In Aim 3, we will establish whether MSA assessment predicts greater care needs and life-relevant disability, examining the Allocation of Caregiver Time Survey, ER visits and hospitalizations, physical activities, and physical performance. MSA assessment is brief and feasible, with potentially greater public health value value than standard generic cognitive screening. We expect that our study will establish a role for objective MSA assessment in geriatric and cognitive care, and we also expect our research results to improve the MSA assessment standard used in pharmaceutical trials that enroll the aged.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSTRACT This application in response to Special Interest Project (SIP) 25-004 will establish a Lifestyle Change Implementation Research Network (LCIRN) Collaborating Center (Component A) and the Coordinating Center (Component B) at the Prevention Research Center at UMass Chan Medical School. The proposed Collaborating Center project will examine the effectiveness and implementation of a commercially available lifestyle change intervention (LCI), Noom, among patients using GLP-1s. GLP-1s have become the new standard of care in weight, type 2 diabetes, and cardiovascular disease management. Clinical guidelines recommend physical activity and dietary-focused LCIs to augment GLP-1s and mitigate side effects. Noom, the first CDC-recognized digital LCI, provides an LCI specific to the needs of GLP-1 users. However, its effectiveness and implementation have yet to be examined. The Collaborating Center project has two aims. In Aim 1, we will conduct a randomized controlled trial of patients using GLP-1s comparing a 4-month digital LCI (n=110) with standard of care (n=110). Primary outcomes include change in moderate-to-vigorous physical activity and Health Eating Index score. Secondary outcomes include frequency of muscle strengthening activities, self-efficacy for exercise and diet, as well as lean body mass, weight, waist circumference, and blood pressure. In Aim 2, we will examine multi-level implementation outcomes with patients, LCIs, providers, and healthcare systems. Guided by the PRISM framework, we will use mixed methods to examine reach, implementation, maintenance, and parity of LCI implementation. In response to Component B, the proposed Coordinating Center is designed to provide an efficient, effective infrastructure that will grow, support and continuously improve a vibrant research network. The network will collectively work to address critical gaps in implementation research of LCIs and implementation of LCIs in real world practice. Aim 1 is to establish and maintain the administrative structure for the LCIRN Coordinating Center and Network. This includes units devoted to Administration and Network Coordination, Communications, Capacity Building, Dissemination and Translation and Evaluation. Aim 2 is to establish a broad, engaged, multi-sector LCIRN membership. We will deploy an intensive, multi-faceted membership drive and provide a versatile membership structure that supports member engagement. Members will be implementers, researchers, payors and other interest holders. Aim 3 is to establish and maintain network-wide activities that build capacity and offer other opportunities for members, partners and the broader field. We will deploy a robust Communications Plan for all members and will support four Communities of Practice (COPs) that provide more in-depth opportunities for capacity building and co-learning in four topic areas: (1) LCI Factors, (2) Patient-Level Factors, (3) Health Care Providers and Systems, and (4) LCI Implementation. Aim 4 is to support the LCIRN Coordinating Centers and build cross-site networking and collaboration. Our work will be informed by Annual Action, Dissemination and Translation and Evaluation Plans.

NIH Research Projects · FY 2025 · 2025-09

Over the last several decades, rates of excessive alcohol use among women (i.e., binge drinking, heavy episodic drinking) have been increasing faster than among men. Alcohol related disease and death are also increasing among women, raising concerns among health professionals, policy makers, and even the White House. Trajectories of binge drinking in young adulthood have shifted; young people are beginning to binge drink later, faster, and longer into their twenties. Women are steadily and significantly increasing their alcohol use into their 30s and 40s, but men’s alcohol use either decreases or remains stable as they age. Globally, men drink more than women, but the difference in those rates vary over time and by contexts. Biological differences between men and women and well-established risk factors of excessive alcohol use are unable to sufficiently explain developmental, cultural, and historical trends in alcohol use. Unique social-cultural and psychological factors contributing to differential trends of alcohol use between men and women in middle-age have yet to be fully delineated. In the U.S., cultural changes have elongated the period of young adulthood (i.e., emerging adulthood) and middle-age is increasingly when adults are getting married and having children. Women are participating in the workforce more than ever before yet are still responsible for the bulk of childcare and household duties. Compared to men, women experience more of the “mental load” associated with these tasks and social roles, are more susceptible to resulting stress and depression, and are more likely to self-medicate their stress and depression with alcohol and other drugs. This study will use the National Longitudinal Study of Adolescent to Adult Health (Add Health) to assess whether and how occupancy of multiple social roles, stress, and negative affect influence excessive alcohol use in two unique stages of life and how those factors form differential pathways to excessive alcohol use (binge drinking, average number of drinks per sitting, and frequency of drinking in past month) among women compared to men. Aim 1 will compare rates of excessive alcohol use between men and women in young adulthood (ages 24-32) and early middle-age (ages 33-43). Aim 2 will delineate the role of these social and psychological factors (stress, negative affect, and occupancy of multiple social roles) as predictors of excessive alcohol use in young adulthood and middle-age and determine if the influence of these predictors is moderated by sex. Aim 3 will characterize mediating and moderating pathways between these hypothesized risk factors and excessive alcohol use using structural equation modeling and traditional moderator analyses. Results will fill critical gaps in our knowledge about how and why the historical gap between men and women in excessive alcohol use is narrowing and uncover potential targets of age- and sex-specific interventions to reduce excessive drinking patterns in early middle age, especially among women.

NIH Research Projects · FY 2025 · 2025-09

My goal is to become an independent clinical scientist focused on transforming health care systems to prevent perinatal anxiety disorders. This K23 award will provide the advanced research training, protected time, mentorship, and research experience I need to launch the next stage of my career. As many as 1 in 5 perinatal individuals experience an anxiety disorder. These disorders confer significant risks to perinatal individuals and their children and disproportionately impact perinatal individuals with low socioeconomic status (e.g., uninsured, receiving public insurance, or facing housing or food insecurity). A shift in focus to prevent perinatal anxiety disorders before they rise to this level of concern is long overdue. Anxiety Sensitivity Interventions are a promising approach to the prevention of perinatal anxiety. Anxiety Sensitivity Interventions are a cognitive-behavioral-based approach to prevention. These brief interventions (<6 sessions) are designed for prevention by targeting a malleable risk factor for anxiety rather than symptom reduction. While Anxiety Sensitivity Interventions have demonstrated effectiveness in the general population, 1) their application in the perinatal population has not been explored, 2) nor have they been scaled to reach a large population. Traditional, multi-session approaches are not likely to have sufficient reach in the perinatal population. Less than 7% of affected individuals receive treatment for a perinatal anxiety disorder. Evidence-based interventions are even less likely to reach perinatal individuals with low socioeconomic status. Approaches that can reach these groups are needed. In my KL2, we identified key components for adapting Anxiety Sensitivity Interventions. Components included content personalization, additional content describing resources (e.g., doulas), user-provided information, and transparent options for data sharing. We then adapted an Anxiety Sensitivity Intervention for perinatal individuals and for digital health to create the Reaching Calm intervention, a multicomponent digital health intervention to prevent perinatal anxiety in obstetric settings. Reaching Calm includes a 1) digital Anxiety Sensitivity Intervention including SMS text messages and web-based user interface, 2) training for obstetric providers, and 3) implementation protocol for integrating anxiety prevention into the obstetric practice setting. In this K23, we will conduct a pilot cluster RCT to assess feasibility (Aim 1), acceptability, and preliminary effectiveness (Aim 2), and examine user-centered design engagement strategies (Aim 3). We will leverage our community-engaged, user-centered design, and implementation science approach to maximize reach. This work will provide the foundational knowledge and formulate the content needed to prepare an R01 submission to test Reaching Calm in a hybrid effectiveness-implementation trial, with the long-term goal of an intervention that can be sustainably integrated into obstetric settings.

NIH Research Projects · FY 2025 · 2025-09

SUMMARY While current antiretroviral therapies (ART) control viral replication, they are unable to fully restore health or a normal immune status. ART-treated individuals experience several comorbidities, including increased cardiovascular disease, bone disorders, and cognitive impairment. Most importantly, therapy interruption leads to the re-emergence of viral replication and progression to AIDS. Therefore, new approaches aimed at eradicating or functionally curing HIV-1 infection are desperately needed. A promising strategy to eliminate latently infected cells after viral reactivation is the ability of immune cells to mediate antibody-dependent cellular cytotoxicity (ADCC). The RV144 HIV-1 vaccine trial in Thailand elicited only a modest 31.2% protective efficacy. Subsequent analyses indicated that this modest protection was correlated with the generation of IgG antibodies (Abs) with potent ADCC activity with low plasma IgA Abs specific to the HIV-1 envelope glycoprotein (Env). This suggests that ADCC may have contributed to the protection observed in the RV144 trial. But key unanswered questions prevent the development of new therapeutic or prophylactic approaches to specifically utilize the ADCC response: What are the structural and conformational features of Envs that are susceptible to ADCC responses? Specifically, what is the basis for the unique phenotype of Envs from the CRF01_AE subtype of HIV- 1, which predominates the Thai AIDS epidemic, and which are intrinsically susceptible to ADCC? Can this phenotype be induced in other Envs through therapeutic intervention? Answering these questions will prove crucial to the design of improved strategies to eliminate HIV-1-infected cells. The long-term goal of the research proposed here is to inform the development of new strategies for utilizing ADCC to eradicate HIV-1. Our central hypothesis is that the sensitivity of HIV-1 to ADCC is determined primarily by two factors: (1) Env conformation, which dictates the exposure of key epitopes targeted by Abs with potent effector function; and (2) properties of the Ab-antigen complex, including the orientation and flexibility of the Fc domain, which determines engagement with the Fc receptor (FcR). A corollary of this hypothesis is that the intrinsic propensity of Envs of different HIV- 1 subtypes to adopt open conformations contributes to their inherent susceptibility or resistance to ADCC. Determining the molecular basis for diverse ADCC sensitivity phenotypes will inform the development of strategies that manipulate Env conformation to stimulate ADCC. We will accomplish this goal using a multifaceted approach involving biophysical, structural, and virological interrogations.

NIH Research Projects · FY 2026 · 2025-09

Apoptotic Dysregulation Across Cell Types in Aging and Neurodegeneration Aging and Alzheimer’s Disease (AD) involve the progressive loss of brain matter, but this phenomenon is poorly understood. One possible contributor to the loss of neurons and other cells is cell death via apoptosis, the canonical pathway for the controlled elimination of cells experiencing lethal stress. Apoptotic sensitivity can vary widely by age and cell type but has never been surveyed in the various cell types of the brain over the lifespan. The goal of the present proposal is to understand the regulation of apoptotic sensitivity in the brain in normal aging, and the potential dysregulation of this sensitivity in AD and other neurodegenerative diseases. The PI, Dr. Zintis Inde, has identified differences among neural cell types in apoptotic regulation, including age- dependent differences, that might contribute to the pathogenesis of AD and other diseases. Here, he proposes to use functional methods for the measurement of apoptotic sensitivity, along with novel applications of induced pluripotent stem cells (iSPCs), viral vectors, and flow cytometric analysis, to investigate these phenomena. In Aim 1, Dr. Inde will use his lab’s functional methods for the measurement of apoptosis sensitivity, quantifying changes in apoptotic priming across aging cell types in normal and AD model mouse brain tissue. In Aim 2, he will examine the role of apolipoprotein E4 (APOE4), a common risk factor for AD, in mouse and iPSC-derived oligodendrocytes. In the R00 phase, Aim 3 will investigate the interplay between neural cell types in the progression of AD, and Aim 4 will validate and implement a newly-developed method for the in vivo measurement of cell type-specific apoptotic sensitivity. The completion of these aims will provide critical insights into normal aging and AD pathogenesis and provide the basis for a future R01 application studying the mechanisms underlying apoptotic dysregulation in these contexts. The proposed training plan encompasses four training goals. First, training in iPSC models will enable Dr. Inde to study cell death regulation in human patient-derived cells, examining the effect of AD-associated genes. Second, training in the use of adeno-associated virus (AAV) vectors will allow for the study of cell death in intact mouse brain tissue. Third, specialized training in aging biology will provide a rigorous foundation for the proposed experiments and the candidate’s future career. Finally, professional development activities, along with the guidance of an experienced group of mentors, will facilitate Dr. Inde’s transition to an independent faculty position. The training plan and experimental goals are supported by an expert team of collaborators and mentors, including primary mentor Dr. Kristopher Sarosiek, a leading expert in cell death, and co-mentor Dr. Mark Albers, an expert in the clinical and translational study of AD. As a whole, the K99/R00 award will enable Dr. Inde’s development into an independent investigator, applying cutting edge methodologies to elucidate the role of cell death in neurodegeneration and pursue novel therapeutic targets arising from this study.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Gene regulatory networks (GRNs) underpin cell identity, function, and response to external stimulations in human health and disease. Their systematic reconstruction from data not only promises to understand disease mechanisms at the molecular level in basic science, but also directly assists the translational search for therapeutic targets and disease subtypes. An accurate GRN reconstruction requires two driving factors – data and method. For data, single-cell technology has presented a unique opportunity for data volume and cell type specificity, but also suffers from severe challenges in sparsity and scalability. In addition, every GRN is heavily influenced by the genetic and epigenetic variations that differ greatly between individuals and cell types. This limits the utility of most existing GRNs, which were reconstructed from the data of cell lines or very few donors, in primary cells and the whole population. In terms of GRN reconstruction method, causal inference holds great promise in its capacity to accurately identify causation from reverse causation and confounding, and therefore reproducibly estimate perturbation outcomes for therapeutic development. However, mainstream causal inference methods face major challenges in GRN reconstruction, falling short in modeling causal kinetics, feedback loops, measurement noise, and GRN rewiring that are widespread in complex biological systems and measurements. We will overcome the data challenges by reconstructing GRNs for each primary cell type/state using 1) population-scale and 2) atlas-scale single-cell datasets that include numerous individuals of the whole population. This project will be built on and extend our recent novel causal inference framework using stochastic differential equations and probabilistic modeling to address the method challenges. We plan to demonstrate their utility in the therapeutic control of gene expression levels. We will release our methodological advances as open-source software and our causal GRNs as a public resource. We expect them to provide the community with comprehensive molecular network knowledge and perturbation outcome prediction to promote disease mechanism understanding and therapeutic target discovery. This will become a part of our long-term goal in generating a full map of multi-modal causal molecular circuits from publicly available data in single-cell and spatial omics.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT Overdose is the leading cause of death for people reentering the community after incarceration. Yet most individuals with justice involvement fail to receive Medications for Opioid Use Disorder (MOUD), which can cut overdose mortality by up to 50%. Drug courts are an increasingly popular alternative to incarceration and mandate and supervise substance use disorder treatment. And while up to 80% of individuals in drug courts have opioid use disorder, fewer than 15% receive MOUD. Interagency collaborations between drug courts and local MOUD providers are necessary for the referral and maintenance of individuals in drug courts to evidence-based MOUD treatment. Known challenges to drug court-MOUD provider collaborations include negative beliefs, poor communication, a lack of awareness of local MOUD providers, and inefficient referral workflow. To address these challenges to collaboration and access to MOUD, we will adapt, implement, and evaluate a package of implementation strategies titled “Clinical Organization and Legal Agency Alliance Building” (“COLAAB”), which aligns with RFA-DA-25-062 to study implementation strategies improving public health and public safety collaborations. COLAAB uses theoretically grounded implementation strategies, including 1) bringing together drug court staff and MOUD providers for process improvements and coalition-building, 2) developing resource guides and communication aids, 3) conducting educational tours of MOUD agencies and court observations, and 4) using an academic liaison to facilitate interpersonal relationships and improve referral workflow. COLAAB was successfully piloted in three Massachusetts drug courts, demonstrating high acceptability, appropriateness, and feasibility. COLAAB now requires a large-scale, multi-site trial to examine implementation outcomes in a wider range of real-world settings and effects on organizational and MOUD service outcomes. Using the Exploration, Preparation, Implementation, and Sustainment (EPIS) framework, we will conduct a hybrid type 3 effectiveness-implementation mixed methods, stepped-wedge, cluster randomized controlled trial implementing COLAAB in 16 adult drug courts (8 in Massachusetts and 8 in Florida). Our specific aims are: (1) adapt COLAAB for widespread implementation in Aims 2 and 3 using focus groups with drug court staff (n=80) and interviews with MOUD providers (n=32) and drug court clients (n=32), informed by the Framework for Reporting Adaptations and Modifications to Evidence-based Implementation Strategies (FRAME-IS); (2) implement COLAAB and assess primary implementation outcomes of adoption and fidelity, and secondary of acceptability, appropriateness, feasibility, and implementation costs using mixed methods; and (3(a) examine the effect of COLAAB on organizational service outcomes of collaboration, negative beliefs, communication, referral processes, and awareness of providers; and (b) explore the effects of COLAAB on MOUD service outcomes of MOUD engagement. If successful, COLAAB could serve as a model for increasing MOUD access in drug courts nationwide through improving drug court-MOUD provider collaborations.

NIH Research Projects · FY 2025 · 2025-09

Project Summary/Abstract During obesity, white adipose tissue (WAT) expands to store excess calories from the diet. WAT expands either by increasing the size of preexisting adipocytes (hypertrophy) or by increasing the number of adipocytes through the differentiation (hyperplasia). Hypertrophic growth has been associated with hypoxia (low oxygen (O2)) in WAT due to ineffective vascularization during its expansion. Hypoxia in WAT has been linked to insulin resistance, ectopic lipid deposition, and mitochondrial dysfunction, although the mechanisms connecting hypoxia to these effects are not well understood. A major function of O2 is that it serves as the terminal electron acceptor (TEA) in the electron transport chain (ETC), which sustains mitochondrial functions including de novo pyrimidine synthesis, reactive oxygen species production, and ATP generation. However, it is unknown how obesity- induced hypoxia impacts the ETC, and if these changes mechanistically explain adipocyte dysfunction upon obesity. In preliminary work, we discovered rhodoquinone (RQ), a novel mammalian metabolite that functions as an electron carrier in the ETC. The RQ-directed ETC circuit employs fumarate, instead of O2, as the TEA, enabling the RQ circuit to support certain mitochondrial functions in hypoxia. Through lipidomic analysis of tissues from two distinct obese mouse models, we found that RQ levels profoundly and specifically rise in the WAT of obese mice. These data inspired the hypothesis that obese adipose tissue reprogram their ETC to the RQ/fumarate circuit to support mitochondrial functions in WAT during hypertrophic/hypoxic expansion. To address this hypothesis, Aim 1 will leverage primary human adipocytes to explore how the RQ- directed ETC circuit impacts differentiation and lipid droplet formation. This will be achieved using stable isotope tracing to measure lipogenesis and western blotting to monitor signaling cascades associated with lipid droplet formation and turnover. In Aim 2 we will test which ETC circuit (UQ/O2 or RQ/fumarate) is preferentially used in the WAT of lean versus obese mice. To this end, we will perform stable isotope tracing assays and respirometry experiments that distinguish these two ETC pathways in vivo. Moreover, this aim will test the therapeutic potential of reprogramming the ETC to the RQ/fumarate circuit during diet-induced obesity using small molecule analogs of RQ. We will determine the impact of RQ on insulin sensitivity, lipid storage, and metabolic parameters via metabolic cages in lean and obese conditions to reveal if RQ can mitigate obesity-induced metabolic dysfunction. Taken together, this work will explore the role of a novel mammalian metabolite in adipocytes during differentiation and upon obesity induction. Beyond defining the fundamental metabolic changes induced by the RQ circuit in differentiating adipocytes, the proposed research is translational, as it will investigate, for the first time, the therapeutic potential of reprogramming the ETC in diet-induced obesity.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY/ABSRACT Novel, personalized solutions to the substance use disorder (SUD) epidemic are desperately needed, as is a workforce of highly skilled scientists who will develop them. Dr. Carreiro is a physician-scientist who has developed a successful independent research program which takes a personalized medicine approach to address the current SUD crisis. In the current proposal, she will leverage her expertise and existing resources to create unique training opportunities for mentees both locally and through her national mentoring network within the specialty of medical toxicology through three integrated training goals and research aims. The training goals are to 1) Enhance Dr. Carreiro's research program and knowledge in precision medicine for SUD to provide novel and innovative opportunities for mentees, 2) Enhance her capacity to effectively provide mentorship to diverse mentees in digital health for SUD, and 3) Advance her leadership skills to support extension of her mentorship activities in a programmatic fashion. Simultaneously, she will curate opportunities for mentees though three related research aims: Aim 1: Development of a novel wearable sensing system, MINDER, which will continuously monitor physiologic parameters, and use machine learning algorithms to accurately identify buprenorphine adherence. Aim 1a: Curate a high quality annotated physiological dataset of individuals undergoing buprenorphine induction using the wearable MINDER-Band. Aim 1b: Use buprenorphine induction data collected to build the machine learning algorithms and clinical interface for the MINDER system, Aim 2: Model wearable sensor based digital biomarkers of opioid craving in patients receiving medications for opioid use disorder (MOUD, including methadone, buprenorphine and naltrexone) and to develop digital phenotypes. Aim 2a: Model trajectories of craving during initiation of treatment with MOUD and examine mediators and moderators between craving patterns and opioid use outcomes. Aim 2b: Develop clinically informed digital phenotypes that predict MOUD outcomes. Aim 3: Develop clinically informed digital phenotypes that predict adverse outcomes of opioid use. This new study will leverage data from Dr. Carreiro's biometric repository as well as prospectively collected wearable sensor data to develop a digital pharmacovigilance tool (POPi: Pharmacovigilance for Opioid Pre-addiction Identification) to monitor opioid analgesic therapy and support safe prescribing. The activities in this K24 award will allow Dr. Carreiro to continue to dedicate time to mentorship, specifically to focus her efforts on post-doctoral and junior faculty mentees who are pursing K23 pathways and scale up her mentorship efforts on a programmatic level. This experience will ultimately propel mentees' own careers in patient-oriented research, enhance the workforce of experienced clinician-scientists focusing on personalized solutions for substance use disorders, and improve the lives of people with this devastating disease.

NIH Research Projects · FY 2025 · 2025-09

The overdose crisis continues to devastate communities across the United States, with justice-involved individuals at particularly high risk of adverse outcomes. Despite a constitutional obligation to provide medical care, only 20% of jails offer medication for opioid use disorder (MOUD) to those eligible, often with limited access. Rural jails, which comprise 66% of all jails, face significant operational and logistical challenges to implementing MOUD programs, creating variability in access to evidence-based care. This study aims to test the effectiveness of Jail ECHO Operations, Planning, and Strategy (J-ECHO OPS), an innovative implementation strategy designed to increase access to MOUD in rural jails. Using a Type 2 hybrid trial stepped wedge design, we will recruit 40 rural jails nationally. J-ECHO OPS, based on the Project Extension for Community Healthcare Outcomes (ECHO) model, will provide semi-monthly virtual learning sessions, case-based discussions, and peer-to-peer learning to jail leadership, jail staff, and community MOUD professionals. The Exploration, Preparation, Implementation, Sustainability (EPIS) framework will guide system change, while the opioid cascade of care will inform best practices. Aims: (1) Test J-ECHO OPS effectiveness on implementation outcomes: MOUD reach, penetration, adoption, sustainability, and cost. (2) Evaluate J-ECHO OPS impact on workforce-level outcomes: MOUD knowledge, attitudes, commitment, efficacy, readiness for change, negative attitudes, and unfair treatment. (3) Assess J-ECHO OPS effects on organizational climate outcomes (diversion, disciplinary infractions, violence involving staff and residents, and conveyance of drug contraband). The team represents a strategic partnership among UMass Chan Medical School, Project ECHO® Institute at the University of New Mexico, the American Correctional Association (ACA), and the American Association for the Treatment of Opioid Dependence. This multidisciplinary team is well-positioned to test overdose prevention strategies in criminal legal settings, with expertise in implementation science, systems change, and leverages ACA’s unparalleled authority as the primary standard-setting and accrediting body for U.S. correctional facilities, ensuring that our MOUD implementation strategies align with industry best practices and have the potential for widespread adoption across the nation's jails. By fostering intercommunication, professional development, and capacity building among correctional and public health stakeholders, this project aims to improve MOUD implementation in rural jails, potentially preventing thousands of overdoses annually and mitigating measurable variability in health outcomes in this high-risk population

NIH Research Projects · FY 2025 · 2025-09

Individuals with substance use and mental health conditions (COD) are overrepresented in criminal legal (CL) settings, including jails with 50% having a COD. Compared to having a single disorder, those with a COD have more serious criminal histories, adverse childhood experiences, reincarceration, suicide, homelessness, unemployment, poor treatment engagement, and a tenfold increased risk of overdosing within 3 months of release. While treatments exist to address mental health, substance use, CL prosocial thinking, and social service needs separately, there is an absence of comprehensive re-entry approaches that address these needs simultaneously, which can result in care fragmentation, poor treatment engagement, relapses, and a vicious cycle of reincarceration. Maintaining Independence and Sobriety through Systems Integration, Outreach, and Networking-Criminal Justice version (MISSION-CJ) is a promising, cross-disciplinary multicomponent intervention, offering 6 months of COD treatment, prosocial and recovery services, and assertive outreach. Four MISSION-CJ open pilots demonstrated increased treatment engagement, improved behavioral health outcomes, and reduced recidivism for clients with a COD. We also have a MISSION-CJ Manual, Workbook, Treatment Planning Tool, and Fidelity Measure and are ready to test them in this RCT. In response to RFA-DA-25-062, the proposed 5-year study, “Supporting Treatment Access and Recovery in Re-entry (STAR-R),” will randomize 240 people with COD to MISSION-CJ or Peer Linkage Support (PLS). Study aims include Aim 1: Compared to PLS, those receiving MISSION-CJ are hypothesized to show: (1a) greater engagement in treatment (measured by total days participated in each condition), and total community provider linkages sessions); (1b) Reduced substance use (measured by self-report use days); reduced overdose risk (measured by self-report Overdose Risk Questionnaire) / overdoses (self-report and corroborated with surveillance data); and reduced mental health symptoms (measured by self-report mental health symptoms); (1c) less CL recidivism (measured by fewer days in jail and fewer numbers of arrests). Aim 2: Examine mechanisms impacting Aim 1 outcomes. Treatment effects will be mediated by: (i) recidivism risk; (ii) increased affiliations with prosocial peers; (iii) reduced affiliations with antisocial peers; and (iv) increased community integration; and will be moderated by demographic factors and COD severity. Aim 3: To conduct a comprehensive economic evaluation that will (i) estimate the full implementation (start-up and ongoing) costs associated with MISSION-CJ and PLS, and (ii) evaluate the cost-effectiveness of MISSION-CJ compared to PLS, from the healthcare system and societal perspectives. Aim 4. To examine facilitators and challenges of MISSION-CJ implementation via qualitative interviews with participants (n=20) and staff (N=12). This application is responsive to NIDA priorities by proposing to improve treatment within the CL system, and to optimize continuity of care post incarceration, and the JCOIN goals of addressing the intersection of the CL system and the community-based healthcare system. This study is part of the NIH’s Helping to End Addiction Long-term (HEAL) initiative to speed scientific solutions to the national opioid public health crisis. The NIH HEAL Initiative bolsters research across NIH to improve treatment for opioid misuse and addiction.

NIH Research Projects · FY 2025 · 2025-09

Project Abstract Atherosclerotic cardiovascular disease (ASCVD) is a leading cause of mortality in the United States, driven by systemic risk factors such as hyperlipidemia, hypertension, and diabetes. However, arteries show varying susceptibility to atherosclerosis, particularly the right coronary artery (RCA) and the left internal mammary artery (LIMA). The LIMA's resistance to atherosclerosis makes it the preferred graft in coronary artery bypass grafting (CABG) due to its long-term patency. Our research in human and mice perivascular adipose tissue (PVAT) has revealed that PVAT in different vascular regions exhibits distinct functional characteristics, particularly in terms of thermogenic capacity. This project aims to investigate the hypothesis that LIMA PVAT has a more thermogenic and anti-inflammatory phenotype compared to RCA PVAT, contributing to its resistance to atherosclerosis. In our proposal, the first aim is to characterize distinct adipocyte subtypes and intercellular signaling pathways within RCA and LIMA PVAT. The second aim is to explore the differentiation potential of PVAT progenitors and the adipocytes derived from PVAT in the RCA and LIMA. The third aim is to correlate thermogenic and anti- inflammatory transcriptomic profiles with imaging biomarkers of PVAT inflammation and atherosclerosis. Through these aims, we will uncover key molecular traits of PVAT that are important in the development of atherosclerosis. Understanding PVAT's influence on vascular homeostasis across vascular beds will lead to new therapies for diverse cardiovascular conditions, including coronary artery disease, peripheral artery disease and carotid artery disease.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Mentorship plays a critical role in the preparation of early career scientists navigating their careers, an essential step toward building a strong and sustainable biomedical research workforce. This proposal brings together Professional Development Hub (pd|hub) and the Center for the Improvement of Mentored Experiences in Research (CIMER) to develop evidence-based curricula and resources to support career- related mentoring conversations in research environments. We propose an approach that is community-driven, with research advisors (supervisory mentors), students/postdocs (mentees), educators, and scholars of career decision-making coming together into a Mentorship Action Collaborative at various entry points to (a) identify priority competencies that should be addressed to enhance career-related mentorship, (b) highlight local resources already being developed in this area, (c) join advisory development teams to inform resources and curricula as they are developed, and (d) be early adopters beta-testing resources and curricula. This project is innovative in its (1) development of novel curricula and resources for research advisors and mentees to enhance evidence-based practices in career-related mentorship, (2) engagement of diverse stakeholders critical to graduate student and postdoc career development through structured communities of transformation, (3) design of assessment instruments to evaluate career development support through mentoring relationships, and (4) building of synergy between mentorship and other supports to maximize mentee outcomes. Using best practices of dissemination and implementation, we will catalyze dissemination across our broad and widespread networks. This comprehensive plan to enhance training will enable graduate students and postdoctoral scholars to adapt, succeed, and thrive in a strong and diverse biomedical research workforce.

NIH Research Projects · FY 2025 · 2025-09

The cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway is a cytosolic double- stranded DNA (dsDNA) sensing pathway critical for regulating immune homeostasis. A series of gain-of-function (GOF) mutations result in constitutive activation of STING, causing an autoinflammatory disease called STING- Associated Vasculopathy with Onset in Infancy (SAVI). SAVI patients succumb to treatment resistant inflammatory lung disease and respiratory failure. There is little known about the mechanisms by which inflammation occurs. To address the urgent need to develop safe and effective therapies, we have developed a murine model for the most common STING gain-of-function mutation, STINGV154M/WT (VM). These mice recapitulate the lung inflammation exhibited by human SAVI patients. To identify the specific cell types involved in causing lung inflammation, we developed a novel VM conditional knock-in (CKI), allowing specific targeting of the VM mutation to different cell types. We demonstrated that endothelial cell (EC) STING GOF is sufficient in driving bronchus-associated lymphoid tissue (BALT) formation. However, the mechanism of action remains to be elucidated. Moreover, we have previously described SAVI lung disease as independent of type I interferon (IFN) and IRF3, signaling proteins downstream of STING activation. STING activation leads to downstream signaling of other pathways including NF-κB and autophagy. The signaling mechanism causing lung inflammation is also unknown. Additionally, STING GOF in ECs was insufficient to cause the extent of lung inflammation seen in VM mice, suggesting STING GOF in cells other than ECs is required for lung disease. Upon ubiquitous VM expression, we find evidence of fibroblast activation in the lung tissue. Fibroblastic reticular cells (FRCs) are a subset of fibroblasts that define the function and structure of lymphoid organs such as BALT. In addition to ECs, STING is highly expressed in FRCs, yet the role of STING in FRCs and contributions to lung disease is unknown. Thus, we hypothesize that coordinated interactions between ECs and FRCs exacerbate SAVI lung autoinflammation, which is dependent on NF-κB activation. In this proposal, Aim 1 will investigate how STING GOF mutation in ECs initiates immune cell recruitment. Aim 2 will determine the synergistic effects of STING GOF mutation in ECs and FRCs on lung autoinflammation. We propose to utilize in vivo, ex vivo, and in vitro techniques to test our hypothesis. The studies proposed in this application will provide critical insights that will enable us to design the best therapies. Furthermore, these studies will provide an opportunity to study the impact of STING activation on stromal cell types, an area of research that requires further exploration. Our findings will discern the role of ECs and FRCs in VM lung autoinflammation and will broadly provide insight into stromal cell-driven mechanisms of other lung disorders.

NIH Research Projects · FY 2025 · 2025-09

Obesity causes mitochondrial dysfunction, disrupting lipid homeostasis, adipogenesis, and energy expenditure. The mechanisms underlying mitochondrial dysfunction in obesity are not fully understood. The proposed research will test a novel mechanism to restore mitochondrial function and mitigate metabolic disease caused by obesity. This plan centers on rhodoquinone (RQ), a previously unknown mammalian metabolite that my lab discovered in preliminary work to be enriched in mouse and human adipose tissues. RQ carries electrons in the mitochondrial electron transport chain (ETC) on a noncanonical path whereby fumarate, instead of O2, is the electron acceptor (Valeros et al., In revision). The fundamental role of the RQ/fumarate pathway in adipogenesis, and its therapeutic potential in obesity have never been studied. Preliminary data reveal that RQ levels rise specifically in the adipose depots of obese mice, and that providing RQ to differentiating adipocytes drives lipid accumulation. Moreover, treating mice with RQ during diet-induced obesity profoundly reduces fatty liver, suggesting that reprograming the ETC to the RQ/fumarate pathway improves systemic lipid handling. These preliminary data inspired the overarching hypothesis that adipocytes engage the RQ/fumarate ETC as a defense mechanism during obesity, rewiring mitochondrial metabolism to enable storage of excess lipids. Moreover, we anticipate that further elevating RQ during obesity onset will improve lipid storage in adipose tissue, reduce ectopic lipid deposition, and consequently mitigate metabolic disease. To address these hypotheses, the first aim will elucidate the mechanism by which the RQ/fumarate ETC circuit drives lipid accumulation during adipogenesis. Specifically, this aim will establish the point of differentiation that adipocytes reprogram their ETC to the RQ/fumarate pathway and measure its impact on mitochondrial functions. Additionally, this aim will test how RQ drives lipid buildup by monitoring lipolysis, lipogenesis, and fatty acid oxidation on in-house high resolution mass spectrometers. The second aim will test the therapeutic potential of reprogramming the ETC to the RQ/fumarate pathway in obesity. This aim leverages first-in-class genetic and pharmacologic tools developed by my lab that reprogram the ETC to the RQ/fumarate pathway in vivo. Specifically, we will analyze the impact of RQ on insulin sensitivity, glucose tolerance, and circulating triglyceride levels in lean and obese models. Also, whole-body lipid deposition, weight gain, activity, and respiratory rate will be measured in collaboration with experts at the UMass Chan Medical School Metabolic Disease Research Center. Our toolkit uniquely positions us to address the fundamental role of RQ, a previously unstudied metabolite, in lipid homeostasis throughout adipogenesis. Moreover, this study will use these tools to test, for the first time, the therapeutic potential of reprogramming the ETC as a strategy to treat diseases such as obesity. Regardless of the outcomes, the proposed research will lay the foundation for a new field of adipose biology on flexibility in the mitochondrial ETC that will ignite the field for decades to come.

NIH Research Projects · FY 2025 · 2025-08

Project summary Over 300 different chemical modifications on RNA have been identified and this ‘epitranscriptome’ plays important roles in the posttranscriptional control of gene expression. Dysregulation of certain RNA-modifying enzymes including RNA methyltransferases (MTases) is linked to cancer initiation and progression, and new evidence suggests these may provide a new class of druggable targets for treating various cancer types. Transfer RNA (tRNA) functions in deciphering the genetic code in mRNAs via codon-anticodon interactions and is one of the most heavily modified RNA species in cells. The diverse and numerous chemical modifications of the anticodon and main body of tRNAs are critical for tRNA integrity and function, thereby affecting protein synthesis. This project will provide major new insight into the role of tRNA modification reprograming and translational remodeling in cancer and cellular stress responses. RNA mass spectrometry and sequencing approaches will be used to comprehensively catalogue tRNA modification and expression dynamics during melanoma progression and metastasis using both human patient-derived xenografts and clinical samples. An innovative new methodology for enhanced Ribosome profiling will be deployed to correlate RNA modification changes with cancer-associated translational remodeling at codon resolution. Functional screening of all >600 human tRNA genes will be performed to identify individual tRNA genes involved in various stress response pathways in cancer cells, as well as tumorigenesis and metastasis in vivo. The role of several tRNA-modifying enzymes and individual tRNAs in melanoma progression and stress response pathways will be investigated, and RNA mass spectrometry, tRNA sequencing, Ribo-seq, and proteomic studies will be used to reveal the underlying molecular and cellular mechanisms linked to cancer initiation, progression, and metastasis.

NIH Research Projects · FY 2025 · 2025-08

Abstract Aneuploidy, the presence of abnormal chromosome numbers, is a prevalent mosaic genetic anomaly observed at low frequency across all human tissues, with frequency increasing with age. Aneuploidy underlies various human disorders including most forms of cancer, Down syndrome, and miscarriage/infertility. However, relatively little is known about the phenotypic consequences of aneuploidy across different chromosomes and tissues. Over the next five years, our laboratory will address critical knowledge gaps in how aneuploidy alters gene expression and its downstream effects on cellular phenotypes across chromosomes, cell types, and cell states using systems-level, quantitative genetic approaches. Chromosomal copy number alterations (CNAs) impact hundreds or thousands of genes at once, resulting in complex effects within gene regulatory networks (GRNs). We will examine whether CNAs exert phenotypic effects in a cell-type-specific manner, using multiplexed CNA fitness quantification approaches across colon, pancreas, and fallopian tube epithelial cells. These assays will systematically evaluate the fitness impacts of CNAs across the genome, enabling identification of cell-type-specific aneuploidy fitness profiles. This project will leverage human telomerase-immortalized diploid cell models to establish a comprehensive collection of non-lethal aneuploid cell lines from each tissue type, enabling deep investigation into how aneuploidy influences cellular fitness. Additionally, the Watson Lab seeks to better model transcriptomic and phenotypic output of CNAs, since many of the gene expression changes in aneuploid cells affect non-CNA-resident genes. By integrating transcriptomic data with transcription factor regulatory networks, this study will reconstruct tissue-specific GRN models to predict the downstream effects of CNA-driven multigene dosage alterations. These models will be used to explore how CNA-driven signals propagate through regulatory networks, revealing new insights into how CNAs disrupt GRNs to support tumor growth in the case of cancer-enriched CNAs, and, conversely, how GRNs may fail in cases of growth-inhibiting or non-viable CNAs. These studies will thus enhance our understanding of GRN robustness in response to polygenic perturbations. Genome-wide CRISPR screens will be performed to identify essential genes in the context of aneuploidy, with a focus on uncovering synthetic lethal interactions that could be leveraged for aneuploidy-targeting therapies. This comprehensive, systems- level approach will provide unprecedented insights into the mechanisms through which aneuploidy affects gene expression, cellular phenotypes, and disease progression across different tissues.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Advances in medicine have revolutionized treatment strategies for end-stage organ disease, cancer, and autoimmune diseases. Most of these advances focus on manipulating the immune system. Unfortunately, these treatments portend an increased susceptibility to invasive fungal infections. A prime example of these risks is demonstrated by the development of Bruton tyrosine kinase (BTK) inhibitors to treat B cell cancers. The three FDA-approved BTK inhibitors all work by covalently attaching to the active site of this kinase, rendering it incapable of its enzymatic activity. These irreversible BTK inhibitors contribute to elevated incidences of invasive aspergillosis, particularly in the central nervous system. The molecular mechanisms of how BTK inhibition facilitates fungal infection susceptibility remain a significant knowledge gap. We and our collaborators previously demonstrated that BTK inhibition by ibrutinib, acalabrutinib, or zanubrutinib dampens neutrophil effector activity against Aspergillus fumigatus and exogenous TNFα or GM-CSF restores these functions. Furthermore, our preliminary data suggest that tissue-resident immune cells (i.e., alveolar macrophages and microglia) produce less TNFα in the presence of BTK inhibition when infected by A. fumigatus. These data argue that the diminished TNFα or GM-CSF may fail to prime neutrophils treated by BTK inhibitors, which in turn permits the establishment of invasive infection. These clinical observations, coupled with our preliminary data, reveal a previously unappreciated critical role of BTK in the innate immune response to invasive aspergillosis. Our overall testable hypothesis is that TNFα and GM-CSF prime neutrophils to respond to invading A. fumigatus and circumvent BTK inhibitors by activating downstream molecules. To address this hypothesis, we propose the following three specific aims: [1] determine the mechanism of cytokine-mediated restoration of neutrophil effector activity in BTK-inhibited neutrophils stimulated by A. fumigatus; [2] elucidate the impact of BTK inhibition on alveolar macrophages, and [3] define the impact of BTK inhibition on microglia in central nervous system aspergillosis. This work will lead to critical insights into the host mechanisms responsible for the failure to contain Af in the presence of BTK inhibition. Successful completion of the proposed studies will reveal critical insights into the host mechanisms responsible for the failure to contain A. fumigatus in the presence of BTK inhibition and provide actionable targets to improve therapeutic strategies in patients receiving BTK inhibitor therapy.

NIH Research Projects · FY 2025 · 2025-08

Project Summary Frontotemporal dementia (FTD) is the most common form of presenile dementia before the age of 60, and no effective treatments are available. 40% of FTD cases are familial and one of the most common genes with FTD-causing mutations is granulin (GRN), which encodes progranulin (PGRN), a secreted regulatory protein. GRN mutations, such as the most prevalent nonsense mutation c.1477C>T (p.R493X), result in PGRN haploinsufficiency, and attempts at developing small molecule, biologic, and gene replacement therapies have proven challenging due to inherent toxicities that result from PGRN overexpression. Precision genome editing strategies to correct the endogenous gene and restore natively regulated progranulin production, even at <100% editing efficiency, therefore represent a superior therapeutic path. The two best-developed modes of precision editing in vivo are base editing (BE) and prime editing (PE), each with complementary sets of advantages and disadvantages for addressing pathogenic C:G-to-T:A mutations such as GRN c. 1477C>T (p.R493X). In this proposal, we have assembled an experienced, multidisciplinary team to develop a maximally effective therapeutic editing approach to treat GRN-FTD caused by this mutation. We will assess candidate approaches that employ either BE or PE, initially in GRN-FTD patient induced pluripotent stem cell (iPSC)-derived neurons and glial cells to correct GRNR493X, and then compare their effectiveness to prevent disease progression in a humanized GrnR493X mouse that faithfully models some aspects of FTD. Our PE development will also test the feasibility of rewriting larger exon segments, thereby enabling correction of additional mutations from the patient population to be addressed with a single corrective reagent. Safe and effective delivery represents one important challenge in therapeutic genome editing, especially in the central nervous system. We will address this challenge by advancing both viral vector delivery and novel non-viral delivery strategies that we are developing to define the safest and most effective delivery modality for these editing technologies. This work promises to lead to a specific, effective therapeutic intervention for a devastating and currently untreatable form of heritable dementia. Our studies will also advance platform technologies and establish a blueprint for developing clinical genome editing approaches, leveraging these tools for additional mutations that cause FTD and other forms of dementia.

NIH Research Projects · FY 2025 · 2025-08

BOSCO/MASSI – ABSTRACT Profilin-1 (PFN1) is an actin-binding protein that is best known for its role in regulating cytoskeletal dynamics. In 2012, we identified mutations in PFN1 that cause the uniformly lethal neurodegenerative disease amyotrophic lateral sclerosis (ALS), however, the mechanism underlying PFN1-mediated ALS remains poorly understood. To address this knowledge gap, our team has established and/or characterized multiple ALS-PFN1 models, including human induced pluripotent stem cells (iPSCs) and knock-in mice with an ALS-linked PFN1 mutation, both of which allow for investigation of ALS-linked PFN1 at physiological expression levels. Further, we have established methods for isolating recombinant PFN1 variants for structural studies using NMR and for examining the molecular dynamics of PFN1 in complex with relevant binding partners. Through our collaborative research, the Bosco and Massi laboratories uncovered defects in phagocytic vesicular degradation in iPSC-derived microglia cells. The phagocytosis, autophagy and endolysosomal pathways all entail vesicular degradation with a convergence on the lysosome. Notably, our study identified a novel interaction between PFN1 and phosphatidylinositol 3-phosphate (PI3P), a lipid that is essential for autophagy and that is involved throughout the vesicular degradation pathway. Based on these observations, we hypothesize that mutant PFN1 impairs autophagy and vesicular degradation through a gain-of-toxic interaction with PI3P and potentially other PIPs involved in vesicular degradation. The goals of this proposal are to interrogate this hypothesis using PIP-specific probes and modulators in multiple ALS-relevant cell types (Aim 1) and through biophysical binding and structural analyses of PIP/PFN1 complexes within biologically relevant membrane contexts (Aim 2). Our preliminary lipidomics data also revealed lipid dysregulation in ALS-PFN1 mice that may be relevant to disease pathogenesis. Intriguingly, several of these lipids also regulate autophagy. In Aim 3, we will examine potential links between autophagy impairment and specific classes of lipids in models of ALS-PFN1 and TDP-43 that are associated with both ALS and the related disorder frontotemporal dementia (FTD). Collectively, the outcomes from these Aims will provide novel insight into PFN1-mediated ALS and unprecedented structural information on PFN1/PIP complexes. We will also gain broad insight into mechanisms of lipid dyshomeostasis and their link to vesicular degradation in ALS and ALS/FTD, with the goal of identifying interventive strategies that will mitigate lipid dyshomeostasis in these disorders.