Johns Hopkins University

NIH Research Projects · FY 2025 · 2022-02

PROJECT SUMMARY/ABSTRACT In the wake of discouraging results from treatment trials in Alzheimer’s disease (AD), the need to identify novel approaches is urgent. There is emerging consensus that the lack of efficacy in AD clinical trials is attributable to disease heterogeneity. Accordingly, identifying predictors of specific AD endophenotypes that could become targets for intervention is of great interest. Among potential options, compelling evidence implicates the stress response as a promising candidate. Indeed, findings from numerous studies converge on the notion that individuals with a heightened sensitivity to stress are at greater risk of developing AD, suggesting that these individuals may represent a group who could be targeted in AD treatment trials. Translating these findings into clinical application has been hampered, however, by a lack of integration between studies that use rigorous experimental interrogation of the endocrine stress response and studies that include sophisticated characterization of patient samples. We aim to address this crucial gap. In this application, we propose to conduct a prospective study to examine the associations among the endocrine stress response, the negative cognitive effects of acute stress, and subsequent cognitive decline in 60 men and 60 women with mild cognitive impairment due to AD (MCI). For our basic study design, we will induce acute stress with the Trier Social Stress Test (TSST), and then measure the endocrine hormone and cognitive responses. The domains of memory and executive functioning will be the primary cognitive outcomes. Salivary samples collected at fixed intervals throughout the TSST will be used to measure stress hormone response; cortisol will be the primary hormone outcome. We will also examine the influence of the APOE gene and polygenic risk scores for AD and collect blood-based biomarkers associated with AD pathophysiology. Specific Aim #1: To determine the association between endocrine response to acute stress and memory and executive test performance following acute stress in individuals with MCI due to AD. Specific Aim #2: To examine the moderating effect of APOE genotype and polygenic risk score on the association between endocrine response to acute stress and cognitive test performance following acute stress in individuals with MCI due to AD. Specific Aim #3: To determine predictors of cognitive decline and neurodegeneration at 2-year follow-up. Secondary Aim: Conduct the analyses from Specific Aims 1-3 in men and women separately in order to identify sex-specific predictors of stress-induced cognitive impairment and cognitive decline and neurodegeneration after 2 years. Public health significance: Given the many recent failures of amyloid-lowering therapies in AD, it is important to identify new potential treatment mechanisms. The proposed study could lay the groundwork for future AD prevention trials targeting stress vulnerability and HPA-axis reactivity in at-risk individuals.

NIH Research Projects · FY 2026 · 2022-01

ABSTRACT/SUMMARY Venoarterial Extracorporeal Membrane Oxygenation (VA-ECMO) is a life-saving intervention for patients with refractory cardiac failure, and the frequency of its use continues to increase in the United States. However, during ECMO, exposure of blood to the non-biological material in the ECMO circuit and to trauma from the pump or oxygenator initiates the coagulation cascade. Consequently, patients become vulnerable to stroke and require systemic anticoagulation. VA-ECMO carries a risk of arterial circuit thrombosis as well as microemboli from platelet aggregates or damaged blood cells, which is hypothesized to cause acute ischemic stroke in patients on ECMO support. In addition, as VA-ECMO immediately restores perfusion to brain, routine administration of high-percentage oxygen for the first hours after ECMO cannulation is hypothesized to cause acute brain injury and worsening of injured brain with reperfusion injury. This neurologic injury early after ECMO cannulation provides an opportunity for intervention strategy when the relation between acute brain injury and these modifiable risk factors, ECMO circuit clots, and hyperoxia, is established. For many years, I have had a strong interest in the effects of arterial-sided thrombosis and hyperoxia on acute brain injury and neurologic outcome. Additionally, I have extensive research experience in brain injury and neurologic outcome in adult patients with mechanical circulatory support devices. I am the director of adult ECMO research at Johns Hopkins Hospital and am involved with studies that have helped to define the association between left ventricular assist devices and neurologic injury. My interest in mechanical circulatory support devices has led me to address key knowledge and practice gaps in VA-ECMO, the concomitant acute brain injury, and its impact on neurologic outcome. The proposed research addresses two important “modifiable” factors—blood clots and hyperoxia—in adult VA-ECMO patients and their relation to neurologic outcome. I propose to investigate whether arterial-sided ECMO-generated clots are associated with acute ischemic stroke (Aim 1) and the effects of hyperoxia within 24 hours of ECMO cannulation on neurologic outcome at hospital discharge (Aim 2). Should ECMO-generated clots and hyperoxia prove to carry a substantial risk for acute brain injury and unfavorable neurologic outcome, I will propose clinical trials to target these modifiable factors by strategies such as using conservative oxygen therapy after ECMO cannulation and changing anticoagulation approaches to reduce cerebral emboli. This grant will equip me with the content, methodological expertise, and multidisciplinary collaboration required to be successful as an independent NIH-funded, patient-oriented clinician-scientist in collaboration with cardiac surgery, critical care, and neurology. This award will also provide critically novel discoveries that may likely impact care related to arterial-sided ECMO-generated clots and hyperoxia in these vulnerable adult patients with critical illness.

NIH Research Projects · FY 2026 · 2022-01

PROJECT SUMMARY/ABSTRACT This K01 SERCA award will provide protected research time and mentoring to Diane Peters, DVM, MS, PhD as she establishes an independent biomedical research career. Dual-trained as a veterinarian and pharmacologist, Dr. Peters has a strong background in comparative medicine, animal models of human disease, in vivo pharmacology, and protease biology. Through completion of the training and research aims outlined in this proposal, she will expand her knowledge of inflammatory bowel disease (IBD) and build an advanced technical skill set, to include immunofluorescent imaging, CRISPR/Cas9 genome editing and gastrointestinal phenotyping methods, that will form a solid foundation for her planned independent research program. IBD is a chronic condition that negatively impacts patient quality of life and is associated with a high public health burden. There is no cure for IBD and a large percentage of affected individuals are unresponsive to all available treatments. Glutamate carboxypeptidase II (GCPII) is zinc metallopeptidase that is highly overexpressed in the two main subtypes of IBD: Crohn's disease and ulcerative colitis. The promise of GCPII as a therapeutic target in IBD has been demonstrated in multiple independent preclinical studies, which have shown that small molecule GCPII inhibitors have significant anti-colitis activity in three mechanistically-distinct mouse models. While it is apparent that GCPII upregulation is relevant in both human and mouse IBD, knowledge gaps exist regarding its function in disease. This K01 SERCA research will yield critical new data relevant to the biology of GCPII in IBD. Specifically, in Aim #1 GCPII expression will be defined in normal and IBD-affected gastrointestinal tissues of human and mouse IBD. In Aim #2, a novel knock-in mouse will be generated that overexpresses human GCPII in the ileum and colon, followed by longitudinal characterization of barrier function, gastrointestinal inflammation, microbiome composition and visceral pain response. It is hypothesized that the localization of GCPII overexpression will be conserved in human and mouse IBD and that adult mice with forced ileum and colon GCPII overexpression will spontaneously develop colitis. Successful completion of these research aims will (1) increase our understanding of GCPII dysregulation in IBD, (2) identify GCPII+ target cell populations with relevance to ongoing to drug development efforts and (3) yield a novel mouse model of IBD that may have increased similarity to human disease. The proposed research will be supervised by a multidisciplinary team of expert scientists and clinicians including: Dr. Barbara Slusher, Director of Johns Hopkins Drug Discovery, Dr. Pankaj Jay Pasricha, Director of the Johns Hopkins Center for Neurogastroenterology, Dr. Cynthia Sears, Director of the Johns Hopkins Germ Free Murine Core, Dr. Robert Anders, Gastrointestinal Pathologist, Dr. Christine McDonald, expert in Gastrointestinal Barrier Function, and Dr. Thaddeus Stappenbeck, Gastrointestinal Pathologist and expert in Mucosal Immunity, who are well-suited to mentor Dr. Diane Peters in her translational IBD research career.

NIH Research Projects · FY 2026 · 2022-01

Brief Summary The number of mechanically ventilated children has grown 80-fold since the 1990s and ventilator- associated infections (VAIs) are a leading complication. In the absence of a gold standard, there is clinical uncertainty around the diagnosis of VAIs, and endotracheal aspirate cultures (EACs) are commonly obtained as a tool to assess for VAI. Although EAC results are not specific for infection versus colonization, many clinicians interpret bacterial growth in EACs as evidence of infection prompting treatment. Indeed, 50% of antibiotics used in the pediatric intensive care unit (PICU) are for VAIs. Thus, over-use of EACs contributes to unnecessary antibiotic treatment, compounding patient morbidity by promoting antibiotic resistance, antibiotic- associated adverse events, and increased healthcare costs. Diagnostic stewardship programs promoting the judicious use of EACs have significant potential to reduce unnecessary antibiotic use and improve the quality of care for thousands of ventilated children. The goal of this study is to (1) define nationwide EAC practices and identify optimal EAC practices in children with suspected VAIs, (2) assess key clinical outcomes after a pilot program implementing interventions to improve EAC practices, and (3) identify facilitators and barriers of EAC diagnostic stewardship programs to inform reproducible implementation strategies. Dr. Sick-Samuels is a faculty member in Pediatric Infectious Diseases at the Johns Hopkins University School of Medicine. She is committed to conducting patient-oriented research to develop, implement, measure the impact of, and disseminate diagnostic stewardship programs to improve the care of complex hospitalized patients. During this career-development award, she seeks to (1) develop expertise in implementation science and patient safety and quality research, (2) analyze clinical outcomes used in patient safety and quality research, (3) apply qualitative research methods to implementation science, and (4) prepare to conduct a multicenter hybrid effectiveness-implementation trial. Her primary mentor, Dr. Aaron Milstone, has extensive experience in clinical research and trials dedicated to prevention of antibiotic-resistance and healthcare- associated infections in children. Her co-mentors, Dr. Sean Berenholtz, is a national and international leader in patient safety and quality and prevention of ventilator-associated infections, and Dr. Jill Marsteller is a national expert in implementation science researching determinants of successful implementation and dissemination of evidence-based practices. Her advisors and collaborators have expertise in critical care medicine, infectious diseases, patient safety and quality, implementation science, human factors engineering, and biostatistics. This work will prepare her to develop an R01 proposal to conduct a multicenter effectiveness-implementation trial of EAC diagnostic stewardship. With this mentored research, the resource rich environment of Johns Hopkins University and the protected time to complete her training goals, Dr. Sick-Samuels will become an independent investigator leading diagnostic stewardship and implementation science research.

NIH Research Projects · FY 2025 · 2022-01

Project Summary/Abstract: Heart failure is a debilitating disease. A growing body of evidence indicates that inflammation plays an important role in the development and progression of this disease. However, the development of immunomodulatory based treatments for heart failure has, so far, been mostly unsuccessful. The PI recently described a population of circulating B cells that adheres to the myocardial endothelium and found that B cell deficient animals have alterations in cardiac structure and function. Moreover, he found that, in rodents, small molecule-mediated modulation of myocardial B cells improves cardiac function after heart attacks. These findings, together with emerging evidence from other research groups, suggest that B cells might be a powerful target for the development of immunomodulatory therapies to prevent and treat cardiac dysfunction. However, our current understanding of myocardial B cell biology is critically lacking. Heart damage triggers a local and systemic inflammatory response characterized by recruitment of inflammatory cells to the injured heart and rapid changes in the spleen. Currently, it is unclear if and how B cells are recruited into the injured myocardium. Moreover, even though B cells account for about half of all the cells in the spleen, and splenic inflammatory changes induced by heart damage have been shown to play a critical role in the progression of heart failure, it is unclear if B cells play a role in the splenic immune response triggered by heart damage. In addition, it remains unknown whether B cell mediated antigen presentation (that together with antibody production and cytokine secretion is one of the 3 prototypical functions of B cells) plays any role within the inflammatory response triggered by cardiac injury. Finally, at an even more basic level, it is unknown how B cells adhere to the endothelium of the uninjured heart. Here, the PI proposes to fill these gaps in knowledge by testing the hypothesis that circulating B cells bind to the myocardial endothelium through specific adhesion molecules and, in response to myocardial injury, enter the myocardium through a CXCL13-CCR5 dependent process, are activated and proliferate in an antigen independent manner, and recirculate between the heart and the spleen to amplify the inflammatory reaction elicited by cardiac damage via MHC-II mediated antigen presentation. The long-term goal of the PI is to use the knowledge gained to facilitate the development of B cell-targeted therapies for heart failure. The PI has been a pioneer in the study of the interaction between B cells and the heart. He is a junior investigator, but he has mastered all the techniques needed to test this hypothesis, he is supported by several seasoned scientists that are co-investigators or collaborators in this proposal, and works within a highly collaborative research environment focused on excellence and innovation. He, therefore, is aptly qualified to effectively test this innovative hypothesis and address, within the timeline of this proposal, the critical knowledge gaps described.

NIH Research Projects · FY 2026 · 2022-01

ABSTRACT Respect for all patients as persons is a core principle of ethical clinical practice, which is embodied in the ways clinicians communicate about patients. Although medical records are an integral method of communicating about patients, few studies have evaluated patient records as a means of communicating attitudes of respect or disrespect from one clinician to another. Our own preliminary work has found that language used in medical records has a direct influence on subsequent clinicians who read the notes, both in terms of their attitudes towards the patient and their medication prescribing behavior. This is an important and overlooked pathway by which attitudes can be passed from one clinician to another and affect the quality of care patients receive. Our overarching goal is to improve respect for patients and thereby enhance the quality of patient care. The specific aims of this proposal are: (1) to define and develop a taxonomy of language in medical records using qualitative analyses with input from clinicians and patients across 5 different medical specialty areas, (2) to measure the use of different types of language in medical records using natural language processing (NLP) that can efficiently detect and quantify positive and negative language in electronic patient records, and then (3) to increase respect in healthcare environments using proven implementation science (IS) strategies (stakeholder engagement, education, audit and feedback, and champions) to motivate and enable health system change. Every encounter with a patient is documented in the medical record. Because language may have a powerful role in influencing subsequent clinician attitudes and behavior, attention to the language used in medical records could have a large impact on the promotion of respect for all patients.

NIH Research Projects · FY 2025 · 2022-01

Project Summary / Abstract Purpose: The REpeated ASSEssment of SurvivorS in ICH study will conduct long-term cognitive, functional, and neuropsychiatric performance assessments to determine if evacuation of spontaneous intracerebral hemorrhage (ICH) reduces the risk of later cognitive decline in the ageing brain. This study will compare rates of cognitive decline under two treatment strategies for intracerebral hemorrhage: the use of minimally invasive surgery with two similar techniques as performed in the recently completed MISTIE III and ENRICH trials, and the current standard of care using data from both controls in MISTIE III and ENRICH and comparative data from The Ethnic/Racial Variations of ICH (ERICH) study (U-01-NS067963) extended into the ERICH- Longitudinal study (R01-NS093870) which followed over 900 of the cases with serial cognitive examinations. Rationale: Intracerebral hemorrhage has the highest disability rate among stroke survivors. ICH survivors are at particularly high risk for progressive cognitive impairment which is strongly associated with greater hematoma volume, but also with cerebral amyloid angiopathy. Compared with standard of care, minimally invasive surgery with effective hematoma volume reduction may improve long-term functional outcomes while also reducing mortality. As such, reducing hematoma volume after ICH may reduce the risk of post ICH cognitive decline. Design: REASSESS ICH is a longitudinal structured serial telephone interview follow-up plus one-time in- person visit of an anticipated 359 ICH survivors enrolled in MISTIE III (2013-2017) or ENRICH (2018-2022). Cognitive and functional outcome data will be compared with up to 900 patients enrolled in ERICH-L, to determine if surgical ICH reduction leads to reduced risk of progressive cognitive decline. Primary Aim 1: To determine if surgical clot reduction after ICH reduces the risk of progressive cognitive decline. Hypothesis: The final residual volume of ICH will correlate with risk of cognitive decline after controlling for age, sex, initial volume of ICH, leukoaraiosis, APOE genotype, and hypertension treatment among operated and non-operated survivors of MISTIE III/ENRICH and survivors of ERICH, and effective clot reduction (<20 mL end of treatment volume), will be associated with lower risk of cognitive decline compared to non-operated patients. Primary Aim 2: To determine if there is a long-term benefit in survival and functional outcome from minimally invasive surgery and the interaction with cognitive decline. Hypothesis: Effective clot reduction will be associated with a decreased risk of death/major disability compared to non-operated patients. Exploratory Aim 3: To determine if inflammatory gene pathway expression predicts risk of cognitive decline. Hypothesis: Chronic brain inflammation contributes to progressive cognitive impairment post ICH. Our preliminary data identifies that inflammation appears to occur chronically after ICH; not just acutely. When testing a wide variety of gene expression changes, the context of which pathway is involved is critical to provide context. We will evaluate whether inflammatory pathways in particular predict patients with cognitive impairment independent of gene risk scores for dementia and surgical status.

NIH Research Projects · FY 2026 · 2022-01

Summary Mitochondria are essential organelles that control the life and death of cells. Mitochondria are highly dynamic: They grow, divide, and fuse, and when they eventually become damaged, undergo degradation Mitochondrial division is mediated by a dynamin-related GTPase, DRP1, while fusion is mediated by two dynamin-related GTPases, OPA1 and mitofusin. These GTPases are mutated in human diseases, including neurodevelopmental disorder, Charcot-Marie-Tooth neuropathy, and optic atrophy. Altered activities of these proteins have also been linked to metabolic syndrome, cardiovascular disease, and age-related neurodegeneration. My laboratory’s goal is to decipher the molecular mechanisms that control mitochondrial structure and translate the fundamental biology to disease interventions. In the past two decades, we have identified and characterized the three essential GTPases in the core reactions of membrane fusion and division. The roles of mitochondrial dynamics are ever-expanding, and now include size control of mitochondria, their distribution and turnover, and differentiation of neurons, cardiomyocytes, stem cells, and immune cells. Most recently, it became evident that the mechanisms of mitochondrial division and fusion are much more complex than initially imagined, involving inter-organelle interactions and a feedback response that monitors and tunes their balance. The emerging new biology is transforming the field of mitochondrial structure and dynamics. In the next 5 years, we will address the important questions raised by this intellectual evolution. First, to our surprise, we found that DRP1 shapes the endoplasmic reticulum (ER) into tubules that form contract sites with mitochondria. DRP1-produced ER-mitochondria contact sites strongly promote mitochondrial division. We will investigate how DRP1 creates ER-mitochondria contact sites that specifically function in mitochondrial division, associates with the ER, and deforms the ER membrane. Second, we discovered a physiological pathway of mitochondrial turnover via DRP1-controlled, Parkin/PINK1- independent mitophagy in mice. This pathway’s most upstream event is to recognize and mark damaged mitochondria by ubiquitination of mitochondrial proteins. Our initial experiments suggested that ubiquitination occurs in two phases – reversible initiation and committed amplification. We will determine what ubiquitinates mitochondria in each phase, and how the ubiquitin ligase complexes recognize and label damaged mitochondria in vivo. Third, we found the first example of a stress response (MitoSafe) that senses and adjusts the mitochondrial structure by controlling the balance between fusion and division. We will explore the molecular basis of MitoSafe and its physiological roles in mice. The MIRA grant will enable us to discover the new logics of mitochondrial structure and its physiological role and regulation in vivo.

NIH Research Projects · FY 2026 · 2021-12

Project Summary/Abstract The global burden of mycobacterial infections remains staggering, with over 10 million new tuberculosis (TB) cases and 1.2 million deaths in 2019, and a recent rising incidence of nontuberculous mycobacteria (NTM). Treatment of both TB and NTM is hindered by the need for prolonged combination antibiotics, drug resistance, toxicity, drug-drug interactions and sub-optimal efficacy. After decades without many new agents, there are now concerted efforts to identify and integrate novel drugs, optimize existing drugs, and augment host immune response. Clinical pharmacology, when integrated throughout drug development, defines critical relationships between drug exposure and effect (or toxicity), identifies key interactions, and enhances treatment success. However, to date, there is a dearth of physician scientists with dual training in infectious diseases and clinical pharmacology who can champion these drug development efforts, especially for NTM. In this proposal, the candidate will apply clinical pharmacology tools to the problem of mycobacterial therapeutics and utilize the resources and framework of three funded clinical trials that are being led by mentors and advisory committee members. Backed by a strong multidisciplinary advisory team with expertise in clinical pharmacology, mycobacterial drug development, pharmacometrics, and mycobacteriology, the candidate will 1) define the pharmacokinetics-pharmacodynamics (PK-PD) of azithromycin for Mycobacterium avium complex (MAC) lung disease by applying the early bactericidal activity (EBA) trial design (Aim 1); 2) identify predictors of toxicity for a novel nitroimidazole antibiotic using PK-toxicodynamic (PK-TD) modeling (Aim 2); and 3) determine the magnitude of drug-drug interaction (DDI) between first-line TB treatment and pravastatin as host-directed therapy (Aim 3). Each one of these aims will provide an opportunity to not only learn but also implement pharmacologic analytical methods for the investigation of both TB and NTM. The candidate will gain skills including population PK modeling, execution of early bactericidal activity trial design, PK-toxicodynamic modeling, and development of a clinical toxicity prediction tool. This K23 Mentored Patient-Oriented Research Career Development Award will serve not only to advance research in mycobacterial therapeutics, but also support the candidate to gain the skills necessary to perform fundamental and advanced clinical pharmacology analyses, lead clinical trials, and form strong multi- institutional collaborations. Upon completion of this K23, the candidate will be well positioned as an independent clinical investigator with expertise in infectious diseases, applied antimycobacterial pharmacology, and NTM therapeutics.

NIH Research Projects · FY 2026 · 2021-12

Project summary Seasonal influenza (“flu”) remains a serious public health threat with the highest burden of severe disease and complications affecting older adults, particularly those over age 75. In addition to vaccine itself, responses to vaccination and vaccine effectiveness in older adults are likely influenced by comorbidity (e.g., frailty), immune senescent remodeling (i.e., immunosenescence and inflammaging), repeated annual vaccination, intra-seasonal immune waning, and virus strain variations both in vaccine formula and in circulation. Since 2014, we have established a study cohort in community-dwelling older adults >75. The cohort has accumulated 815 person-seasons with comprehensive demographic, clinical, functional and laboratory data, as well as banked pre- and post-vaccination serum, plasma, and peripheral blood mononuclear cell (PBMC) samples. We also identified 15 breakthrough flu infection cases with banked post-infection serum, plasma and PBMC samples. Importantly, 20 subjects participated in all 7 seasons, 36 in 6 seasons, 31 in 5 seasons, 16 in 4 seasons, and 165 in 3 seasons or less. Here, we propose to leverage this unique cohort and employ cutting edge immunologic research tools to develop state-of-the-art “immune signatures” reflecting both general immune status (distribution and function of immune cell subsets through high-dimensional flow analysis and RNA-Seq; cytokine profiling) and influenza-specific immunity (breadth and depth of flu-specific T cell repertoire; distribution/function of homotypic/heterotypic anti-flu T cells through flow analysis and scRNA-Seq; deep serological profiling of strain-specific and cross-reactive flu antibodies). Our objective is to characterize immune signatures and their intra- and inter-seasonal changes over time as determinants of vaccine responses and risk of breakthrough infection in older adults >75. Our specific aims are: 1) Characterize seasonal baseline (pre-existing) immune signatures as determinants of vaccine response and how they change over time. We will not only determine inter-season longitudinal trajectory, but also identify specific baseline immune signatures predict responses to vaccination; 2) Characterize seasonal immune signature responses to vaccination as determinants of risk of breakthrough infection and how they change over time. We will evaluate and compare differences and similarities of immune signature responses elicited by vaccination vs natural infection to explore immune mechanisms of vulnerability; and 3) Characterize intra-seasonal waning of immune signature responses to vaccination and its change across seasons through monthly blood sampling until the end of each flu season across multiple seasons. Upon completion, the proposed studies will advance our understanding of immune signatures as key immunologic mechanisms for vaccine responses and risk of breakthrough infection in a typical geriatric population. Ultimately, these studies will help define correlates of protection and develop more effective immunization strategies including a universal vaccine for this highly vulnerable subset of older adults.

NIH Research Projects · FY 2026 · 2021-12

PROJECT SUMMARY/ABSTRACT Jennifer Dantzer, MD MHS is an allergy and immunology physician-scientist at Johns Hopkins University, with a Master's Degree in Epidemiology. This K23 award will provide her valuable training towards her long-term career goal of being an independent investigator focused on patient centered outcomes (PCO) of food allergy treatments. Food allergy is a major public health disorder that affects at least 5.9 million American children and is increasing in prevalence. Historically, food allergy management has been based on strict allergen avoidance and recognition and treatment of allergic reactions. However, the burden of food avoidance and on-going uncertainty over possible accidental exposures can have a significant psychosocial impact on children and parents. The FDA approved the first treatment for peanut allergy, Palforzia, peanut oral immunotherapy (POIT), in January 2020, ushering in a new era for food allergy treatment. While this paradigm shift in food allergy management is monumental, there is significant uncertainty related to benefits/risks to the patient, treatment preferences, and patient-centered outcomes (PCO). This gap in our understanding lead to this project's overall objective to evaluate the patient perceived benefit of peanut allergy therapy and to better understand decision-making with regard to discontinuation and treatment preferences. Aim 1 will examine longitudinal change in child, adolescent, and caregiver food allergy related quality of life in a diverse, real-world population on and off POIT. In addition, the long-term benefit of POIT is dependent upon on-going treatment since stopping treatment causes the majority of patients to lose most or all protection. Aim 2 will use local and national data as well as interviews and questionnaires to determine the rates, potential predictors, and reasons for POIT discontinuation. Finally, Aim 3 will use a mixed methods approach to understand patient and parent preferences for PA treatment. This K23 award addresses the mission of NIAID to better understand and treat allergic diseases by using a novel approach and methods directed towards PCO research. The research will be completed within the highly supportive training environment at Johns Hopkins, with the guidance of a team of expert mentors with relevant expertise (Drs. Robert Wood, MD, Corinne Keet, MD PhD, Albert Wu, MD MPH, Cynthia Rand, PhD and John F.P. Bridges, PhD). The career development plan combines hands-on training, formal didactics, and outstanding mentorship in key areas of statistical analysis, epidemiology, and advanced techniques in PCO research. This plan will ensure a successful transition to independence. This K23 award will provide preliminary data and skills that are highly relevant and applicable to future R01 or U level grants. Findings have the potential to change clinical practice not only related to POIT, but also for future food allergy treatments, through enhanced benefit-risk assessment, best practice guidelines, and patient decision-making tools.

NIH Research Projects · FY 2025 · 2021-12

Project Summary - Overall The mortality rates of breast and pancreatic cancers are intrinsically tied to metastasis. In pancreatic cancer, the 5-year survival rate is only 9% and in ~70% pathological evaluations of the resected tumor, instances of venous invasion are found. Metastasis is a complex multi-step process involving cancer cells, local vasculature, and the surrounding microenvironment at multiple sites. Venous invasion in pancreatic cancer, in which cancer cells gain often invade the portal vein, is an early step in this process and provides the cells a direct path to the liver, the most common site of pancreatic cancer metastasis. As in pancreatic cancer, invasion past normal breast tissue barriers is critically tied to breast cancer outcomes. Most breast tumors can be surgically removed, and so mortality is closely tied to the extent of distant metastasis through lymphovascular invasion. The detection of lymphovascular invasion in a breast tumor correlates with poor prognosis and is not captured in current molecular analyses. The spatial organization of cancer cells, and cellular and stromal components of the tumor microenvironment near and far away from blood vessels is intrinsically three-dimensional, non-symmetric, and highly heterogeneous. In the TECH units of the Johns Hopkins Center for 3D Multiscale Cancer Imaging, we will develop a versatile 3D multiscale imaging method, CODA, which will allow us to probe the phenotypic heterogeneity of tumors from the multi-cm to the micron scale via multiplexing serial imaging. CODA can readily incorporate other imaging modalities to extract high cellular/molecular content from 3D samples. These include immunocytochemistry (CODA+IHC), immunofluorescence (CODA+IF), imaging mass cytometry (CODA+IMC) and spatial transcriptomics/proteomics (CODA+DBiT-seq). These proposed expanded versions of CODA offer a unique opportunity to produce new 3D multi-omic maps of human PDAC and breast tumors near and far from blood vessels. CODA and its integrated versions CODA+X will be tested in the RTB units of the Center in both human/mouse tissue samples and organoids, in breast and pancreatic cancer. Results from these test beds will provide novel mechanistic insights into venous invasion in breast and pancreatic cancer. Exploiting our extensive experience in data dissemination, we will make the large datasets and software produced by our CODA+X platforms and software widely available to the larger cancer research community. The units of the Center will be co-led by Johns Hopkins/Yale engineers, scientists and physicians. Substantial additional support will be provided by the Johns Hopkins U., the JH School of Medicine, the Institute for Convergence, the Department of Pathology, and the Whiting School of Engineering.

NIH Research Projects · FY 2025 · 2021-12

PROJECT SUMMARY Adipose oxidative metabolism is central to human health. Defects in adipocyte mitochondria are linked to adipocyte dysfunction and insulin resistance, whereas lifestyle interventions and pharmacological agents that promote adipocyte oxidative capacity promote metabolic health and insulin sensitivity. Studies over several years have identified important regulators of adipocyte oxidative metabolism, such as members of the PGC-1 coactivator family, and nuclear receptors of the PPAR (peroxisome proliferator-activated receptor) and ERR (Estrogen-related receptor) subfamilies. These transcription factors exert their effects on gene expression and cellular function by both direct and indirect regulation of hundreds of genes that coordinately promote mitochondrial biogenesis and oxidative metabolism. Mining of genes regulated by PGC-1s and ERRs, coupled to bioinformatic approaches to determine associations of PGC-1/ERR targets with PPAR pathways and metabolism, has led us to identify a new and poorly characterized protein, Mcrip2, as highly associated with adipocyte oxidative metabolism. The premise of this proposal is that Mcrip2, a protein of unknown cellular and molecular function, that has not been linked yet to metabolism or physiology, is induced by PGC-1, PPAR and ERR factors, acts to enhance basal and adrenergically stimulated expression of oxidative metabolism genes, and is thus a critical element of the regulatory networks that control adipocyte oxidative metabolism. Interactions of Mcrip2 with proteins involved in mRNA processing and turnover suggest that Mcrip2 exerts its function by regulating gene expression at the post-transcriptional level. Notably, we know little about mechanisms controlling post- transcriptional steps in adipocyte oxidative metabolism pathways. The proposed work will define the role of Mcrip2 in adipocyte basal oxidative metabolism and adrenergic responses, using gain- and loss-of function approaches in primary brown and inguinal adipocytes, and delineate the level at which Mcrip2 impacts gene expression. It will also determine the physiologic significance of Mcrip2 for mitochondrial function and adaptive thermogenesis, using a mouse model. Finally, the studies will elucidate the mechanism by which Mcrip2 impacts adipocyte biology, by defining Mcrip2 protein domains required for function and critical Mcrip2 interacting partners in basal and adrenergically stimulated adipocytes. In sum, the work will give first insights into post-transcriptional regulation of adipocyte oxidative function, and may suggest new targets and avenues for therapeutic intervention in diseases that can benefit from increases in oxidative capacity, such as obesity and obesity-related diseases.

NIH Research Projects · FY 2026 · 2021-12

The Children's Environmental Health Research Centers have made significant contributions to defining environmental risk factors and developing interventions to reduce risk. Successful translation of this work has the potential to amplify discovery to improve the lives of millions of infants and children. The Center for Childhood Asthma in the Urban Environment was established in 1998 as one of the original Centers and has focused on childhood asthma and respiratory disease for over twenty years. The BREATHE (Bridging Research, Lung Health, and the Environment) Children's Center will translate findings, based on our own work and work from other centers, regarding environmental exposures that impact children's respiratory health and interventions to reduce risk. The NIEHS translational framework will be applied to put science into practice with novel, evidence- based communication strategies that inform and engage students, community members, health professionals, and policymakers to reduce harmful exposures and promote children's respiratory health. The Program will be overseen by the Administrative Core (Director: Dr. McCormack) which will provide resources, oversight, and support to successfully translate findings regarding children's environmental respiratory health from science to practice and policy. The goal of the Translation Core will be to develop novel communication tools that build on local, state, and national partnerships. The Kids BREATHE Lung Health Dashboard will display relevant, actionable information with map-based visualization strategies and storyboard techniques to engage the general public. The dashboard will synthesize decades of children's environmental health research findings to communicate knowledge about hazardous exposures, associated health risks, and the potential benefits of interventions. The Lung Health Ambassadors Program (LHAP) curriculum will serve as the foundation upon which we will build a curriculum focused on children's respiratory environmental health. LHAP is a school-based health education program with a goal to educate and empower youth to be advocates for lung health that was established in 2018 in partnership with the Baltimore community. The LHAP curriculum will be tailored to reach students, community members, and healthcare professionals and LHAP graduates will be channeled into a new Lung Health Corps, a volunteer network developed to engage key partners at the local, state and national level in a coordinated advocacy program. Finally, the BREATHE Children's Center's mission includes a deep commitment to training the next generation of scientists engaged in children's environmental health research. The Development Core will expand the engagement of early career investigators conducting children's environmental health research through pilot projects and development activities. Complementary pilot programs will provide a mechanism to engage early investigators, to promote agility in responding to emerging environmental exposures and to adapt new approaches to translating scientific discovery into action and policy.

NIH Research Projects · FY 2026 · 2021-12

Project summary Our proposed study aims to address the function of LIN28B and follistatin in supporting cell reprogramming and hair cell regeneration in the murine cochlea. Loss of auditory hair cells (HCs) due to disease or trauma is permanent and is a leading cause for hearing impairments and deafness in humans. Immature auditory supporting cells (SCs) do regenerate HCs in response to damage but their ability to regenerate lost HCs rapidly declines as SCs undergo maturation and little to no HC regeneration is observed in adult animals. We recently uncovered that the RNA binding protein LIN28B and its paralog LIN28A control the regenerative capacity of cochlear SCs in neonatal cochlear organoids and explants. Whether LIN28A/B has a similar role in cochlear HC regeneration in vivo has yet to be tested. Furthermore, our recent in vitro studies suggest that LIN28B promotes HC regeneration through reprogramming SCs into progenitor-like cells and that such state transition can be further enhanced by the co-activation of the Activin antagonist follistatin. However, whether SCs truly activate a transitional progenitor-like state during HC regeneration and if so, how LIN28B and FST may influence such state transition are still unresolved. In our proposed study we will use mouse genetic tools to address whether LIN28A/B regulates spontaneous cochlear HC regeneration in the immature cochlea in vivo (aim1). Furthermore, we will use single cell RNA sequencing and single molecule FISH to determine whether LIN28B reprograms cochlear SCs into progenitor-like cells during HC regeneration (aim2). Moreover, to establish how LIN28B and FST enhance SC reprogramming and subsequent HC regeneration we will manipulate the function of potential LIN28B and FST effector genes using lentiviral overexpression and CRISPR-Cas9-mediated knockout strategies in cochlear organoids (aim3). Finally, we will determine whether co-expression of LIN28B and FST with Atoh1 enables SCs to regenerate cochlear HCs in the mature cochlea in vivo (aim4).

NIH Research Projects · FY 2026 · 2021-12

Bipolar androgen therapy (BAT) is a paradoxical approach for the treatment of castration-resistant prostate cancer (CRPC) whereby testosterone levels are rapidly cycled between supraphysiologic and castrate concentrations. Understanding how BAT works at the molecular and cellular levels might help in rationally combining BAT with other agents to achieve increased efficacy and tumor responses. Previous observations suggest that supraphysiological testosterone (SupraT) induce DNA double strand breaks (DSB). It has been speculated that if left unrepaired, DSBs may lead to cellular crisis and apoptosis. In this proposal, we provide novel evidence that unrepaired DSBs induced by SupraTs are routed to the autophagosomes where they activate cytoplasmic nucleic acid sensors that trigger the downstream interferon stimulated genes (ISGs) and innate immune pathways. Based on our preliminary data, we propose a novel idea that: i) Unrepaired DSBs induced by SupraTs are routed for specialized autophagic degradation termed nucleophagy; ii) SupraTs induced autophagosomal DNA can activate cytoplasmic DNA sensing pathways~ specifically the nucleic acid sensing pathway (cGAS-STING and RIG-I pathway); iii) Activation of nucleic acid sensing pathway by SupraTs would be more pronounced in prostate tumors that have DNA repair defect; and iv) BAT might activate innate and adaptive immune cells specially, in a subset of patients having DNA repair defect. In this proposal, we will determine the role of nucleic acid sensors in mediating immune signaling by SupraTs in PCa. Utilizing tumor biopsies from PCa patients receiving BAT, we will evaluate whether nucleic acid sensor mediated innate immune signaling serves as a molecular determinant of treatment response. To test our hypothesis, we will utilize several innovative tools and resources including CRISPR-Cas9 generated knockout cellular models, GeoMx Digital Spatial profiling of immune landscape in the tumor microenvironment, unique patient derived prostate cancer xenografts models, a humanized mouse models that has functional innate and adaptive immune cells, and serum and tumor biopsies from patients receiving BAT. We think these unique resources position us well to undertake the proposed work with immediate clinical impact. We have assembled a team of experts in basic and clinical prostate cancer biology and immunotherapy who will provide their unique expertise to successfully accomplish our goals. Successful completion of the proposed work will generate: a) mechanistic insights into modulation of immune response by SupraTs, b) valuable clinical insights into activation of immune cells by BAT, c) novel tumor- and serum-based markers that can be utilized for the development of biomarkers predictive of therapy response, and d) provide a rationale for strategically utilizing BAT to activate immune response and combine it with immunotherapeutics that empower the adaptive immune system such as immune checkpoint blockade and T-cell therapeutics to achieve synergy.

NIH Research Projects · FY 2026 · 2021-12

PROJECT SUMMARY The long-term goal of my laboratory is to elucidate the mechanisms that regulate auditory perception, and how defects in hair cells and auditory neurons cause disease. We propose here to study the mechanisms that regulate the differentiation, connectivity and function of spiral ganglion neurons (SGNs). Our central hypothesis is that SGNs are a diverse group of neurons with different connectivity patterns and functions, and that diversification of SGNs and their connectivity depends on signals provided by hair cells. To test our hypothesis, we will combine studies in genetically modified mice with histology, imaging, genomics, electrophysiological and hearing tests to study SGN connectivity and function. We will map SGN projection patterns, define how mutations linked to deafness affect SGN differentiation and projection patterns, and use genetic manipulations combined with electrophysiology and genomic tools to identify signals initiated by hair cells that affect SGN differentiation. Our preliminary data show the feasibility of our approach, demonstrating that SGNs are a molecularly diverse group of neurons with different projection patterns and that defect in hair cells affect SGN differentiation. We anticipate that our findings will define mechanisms by which hair cells affect SGNs to regulate their diversification into subtypes with different functions and connectivity patterns.

NIH Research Projects · FY 2025 · 2021-11

PROJECT SUMMARY / ABSTRACT The overall objective of this K23 application is to provide support for the additional training and experience that Dr. Joy Wan, a pediatric dermatologist and epidemiologist, needs to develop an independent, patient-oriented research program in pediatric atopic dermatitis and chronic skin disease. Research: Atopic dermatitis, also known as eczema, affects 20% of children and frequently overlaps with the critical period of cognitive development that occurs in the first two decades of life. Although children with atopic dermatitis are more likely to suffer from learning disabilities, inattention, and poor memory, the impact of atopic dermatitis on cognition has not been rigorously examined. This project will: (1) test the hypothesis that atopic dermatitis, particularly when severe or persistent, is both directly and indirectly associated with cognitive impairments in children, and (2) identify modifiable risk factors that mediate the effects of atopic dermatitis on cognitive function. To achieve these aims, Dr. Wan will use data from a longitudinal study of over 14,000 individuals followed from birth until their 20s and also prospectively enroll a cohort of children with active atopic dermatitis. The results of this project will fill critical gaps in knowledge about the impact of atopic dermatitis on cognition and carry significant implications for clinical practice. Candidate: Dr. Wan earned her MD and MSCE degrees from the University of Pennsylvania, where she also completed dermatology residency followed by pediatric dermatology fellowship at the Children’s Hospital of Philadelphia. Dr. Wan is pursuing post-doctoral training in epidemiology with the goal of becoming an R01-funded investigator and leader in pediatric dermatology research. Environment: Dr. Wan will be primarily mentored by Dr. Joel Gelfand, a dermatologist and epidemiologist with expertise in patient-oriented research in psoriasis and atopic dermatitis, and co-mentored by Dr. Susan Furth, a pediatric nephrologist with expertise in chronic disease epidemiology and patient-oriented pediatric research, Dr. Kristin Linn, a biostatistician with expertise in advanced statistical methods, and Dr. Nina Thomas, a pediatric neuropsychologist with expertise in neurocognition research. She will be further advised by a complementary committee of NIH-funded investigators in pediatric dermatology, neuropsychology, epidemiology, and atopic dermatitis. With its robust research infrastructure and rich resources, Penn provides an ideal environment for Dr. Wan’s research and career growth. Career Development: Dr. Wan’s proposal includes advanced training in longitudinal data analysis and causal inference, neurocognitive development, and patient-reported outcome measures, achieved through a combination of didactic coursework, practical research experience, and focused mentorship. The support of a K23 award will enable Dr. Wan to launch a successful career as an independent investigator dedicated to optimizing the health, social, and life outcomes for children with chronic skin disease.

NIH Research Projects · FY 2025 · 2021-09

Idiopathic Normal Pressure Hydrocephalus (iNPH) is a reversible form of dementia and gait imbalance in the elderly that has been treated with surgical ventriculoperitoneal shunting (VPS). Although VPS has been performed for decades, the effectiveness of VPS has not been tested in an appropriately run placebo-controlled clinical trial. Due to the lack of data from placebo-controlled trials, skepticism about VPS in the elderly has significantly limited its use. The primary goal of this research proposal is to gather data with the goal of a definitive answer on the question of whether shunt surgery offers a benefit to patients with iNPH. We will accomplish this through a prospective, blinded, randomized placebo-controlled clinical trial that uses a contemporary FDA approved adjustable shunt system. This valve allows a “virtual off” setting allowing for noninvasive and reversible assignment of patients to treatment with a functioning (active group) or non- functioning (placebo group) shunt. The trial will enroll 100 iNPH patients at 20 sites. Participants selected for shunt surgery will be chosen based on standard and widely used iNPH guidelines. The trial is a delayed treatment paradigm where all 100 patients will receive the same surgery and device, differing only in the initial valve setting (active or placebo). The Primary analysis will be a group comparison of change from baseline in gait velocity at three months after implantation. Secondary analysis will measure balance, cognition, and bladder control. Three months after implantation, all participants in both groups are blindly adjusted to the active setting and followed for 9 months. A secondary goal of the study is to evaluate clinical, imaging, and CSF biomarkers before surgery to identify associations with subsequent shunt response. MR imaging and extended neuropsychological testing are also repeated after shunting to evaluate specific anatomical and cognitive domain changes that may be associated with gait changes. We expect that if we demonstrate the effectiveness of shunting in iNPH we will establish an effective treatment and facilitate an increase in appropriate iNPH shunting. In addition, the study of patient biomarkers associated with successful outcomes will allow future testing of algorithms for more accurate and efficient patient selection. Identification of subgroups of iNPH patients with improvement may also be suggested. Data from this controlled trial can ultimately benefit the 300 to 700 thousand iNPH patients in the US to receive an effective surgical treatment. If data shows that shunting is found to be ineffective in the placebo group, elderly patients will be spared the potential morbidity and cost of an ineffective brain surgery.

NIH Research Projects · FY 2025 · 2021-09

There is no available treatment for vision loss in dry age-related macular degeneration, a leading cause of blindness in adults over 55 years of age. Vision loss occurs when retinal pigment epithelium (RPE) and cone photoreceptors degenerate, so transplanting them is a potential treatment to restore vision. Using a novel mouse cone and RPE transplantation model system to study cone-RPE interactions in a regenerative paradigm, we will characterize donor cone maturation over the long term, reveal age-related impact on regeneration efficacy, and identify cues in specific recipient cell types that promote functional vision repair.

NIH Research Projects · FY 2025 · 2021-09

Abstract Older adults are less able to maintain homeostasis or sustain physiological constancy and, although diabetes is common in old age, glucose instability is an underappreciated concern. Diabetes is strongly linked to dementia risk and there is growing evidence that glycemic variability contributes to risk of Alzheimer's disease and related dementias, independent of average glucose or hemoglobin A1c (HbA1c). Wearable continuous glucose monitoring technology is an opportunity to rigorously characterize glucose patterns in older adults across the glycemic spectrum (from no diabetes, to prediabetes, to diabetes) and evaluate associations with neurocognitive outcomes. The proposed project will 1) characterize midlife- and late-life risk factors for specific glucose patterns in old age (glycemic variability, hyperglycemia, and hypoglycemia); 2) evaluate associations of glucose patterns with concurrent symptoms (dizziness, difficulty concentrating, vision disturbances); 3) examine associations of diabetes, glycemic control, and glucose patterns with brain MRI measures and cerebral amyloid beta deposition via florbetapir PET imaging; and 4) evaluate the associations of disordered glucose patterns with progression of cognitive decline and development of mild cognitive impairment or dementia. Completion of these aims will move the field forward in our understanding of the link of diabetes and continuous glucose monitoring-defined glycemic instability with neurocognitive outcomes in older adults.

NIH Research Projects · FY 2024 · 2021-09

Abstract / Summary Currently, supervised surgical training provides only a small fraction of surgical experience in the career of a practicing surgeon. Surgeons’ skill develops throughout their career. Surgeons benefit from supervised feedback from experts during training, but they lose such structured and specific feedback once they begin independent practice. Surgical skill is associated with patient outcomes. Therefore, supporting surgeons’ continuous professional learning through automated structured resources can improve patient care. The status quo for surgeons in practice is to measure patient outcomes or other process of care variables as indirect measures of their skill. These measures do not inform surgeons how to improve. The goal in this project is to develop tools to analyze videos of the surgical field to provide surgeons with unbiased skill assessments and specific feedback on how to improve. This project includes integration of these tools into a personalized surgical learning platform and evaluation of its effectiveness for surgeons’ skill acquisition. To achieve this goal, this project includes a multi-disciplinary team to include expertise in ophthalmology, surgical education, surgical data science, computer vision, machine learning and deep learning, statistics, and human-computer interaction. The video analysis tools developed in this project will enable the following for cataract surgery, one of the most common surgical procedures in the U.S. and across the world: 1) objective assessments of surgeons’ skill; 2) provide surgeons with specific feedback on how to improve that is personalized given their past performance; and 3) preliminary evidence of effectiveness of a personalized learning platform for surgeons’ skill acquisition. The anticipated impact of our work is to create a pathway in which the surgeon is incentivized to see themselves and their performance as part of the process of improving outcomes and value in care, and institutions have access to objective tools to create reproducible standards for surgical competency.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY Aging is the greatest risk factor to α-synucleinopathy, a group of neurodegenerative diseases with severe cognitive impairmentand progressive motor dysfunction and dementia, such as Parkinson's disease (PD), dementia with Lewy bodies (DLB) and Parkinson's disease with dementia (PDD) and half of Alzheimer's disease patients (AD). Dementia is a common symptom in α-synucleinopathies: DLB is the 2nd most common dementia after Alzheimer's disease (AD) accounting for 30% of dementia cases; Around 30% of AD cases also suffer from α-synucleinopathy resulting in a more rapid and severe cognition decline than AD alone. PD is the 2nd most common neurodegenerative disease, and greater than 50% of PD cases develop PDD. In addition to cognitive and memory dysfunctions, patients with dementia also suffer from anxiety, depression and mood swings. Although α-syn pathology is highly associated with dementia, the underlying aging-related mechanism driving the pathogenesis and contributing to their progression is not known and there is no available disease modifying therapy yet. Based on substantial postmortem analysis, Braak et al. demonstrated that α-syn pathology spreads in a stereotyped fashion from the vagus to the brain, which may initiate in the gastrointestinal tract. Particularly, nearly all the DLB and PDD cases present with α-syn pathology in the gut. Both clinical and experimental observations support that pathogenic α-syn spreading is a master trigger that drives α-synucleinopathy. In our gut-brain α-synucleinopathy (GBAS) mouse model, gut-injection of pathogenic α-synuclein (α-syn) can recapitulate α-syn pathology gut-brain spreading and cognitive impairment. In our preliminary studies, heterochronic blood exchange (HBE) from young mice inhibited pathogenic α-syn transmission and neuroinflammation in aged mice, suggesting an HBE-transferred phenotype that may effectively inhibit α-synucleinopathy. We identified lymphocyte-activation gene 3 (LAG3)1, a major receptor of pathologic α-syn transmission. To identify the mechanism underlying rejuvenation and accelerated aging event, we further identified two novel LAG3-related and aging-regulating proteins that can mediate pathogenic α-syn transmission. Our studies support the feasibility to modulate plasma levels of FGL1 and sLAG3 in aged mice by the HBE approach. To determine the underlying mechanism if FGL1 and sLAG3 in the plasma are molecular mediators essential for the inhibitory effects of HBE on α-synucleinopathy an d related cognitive impairment, we have established a rigorous and robust experimental system combining the HBE approach, the GBAS model, genetically engineered mice without these factors, and recombinant FGL1 and sLAG3 proteins, for comprehensive gain- and loss-of-function analysis. Our Central Hypothesis is to identify the underlying mechanism that HBE inhibits α-synucleinopathy and related cognitive impairment through FGL1 and sLAG3. FGL1 functions as a rejuvenation factor to inhibit pathologic α-syn spreading in the gut-brain axis and alleviate consequent neurodegeneration, neuroinflammation, and cognitive impairment. sLAG3 acts as an age-acceleration factor contributing to the pathogenesis and with antagonistic function to FGL1. Strikingly, human postmortem evidence shows that α-syn pathology is observed first in the gastrointestinal system and then spreads to the brain in a stereotyped fashion. Recently, our collaborator Dr. Dawson developed a novel Gut-Brain α-synucleinopathy (GBAS) model, a sporadic α-synucleinopathy model recapitulating pathologic α-syn spreading among multiple organs and brain regions in patients. However, it remains largely unknown how aging-associated blood-borne components modulate α-synucleinopathy. As the foundation of this project, we identified lymphocyte-activation gene 3 (LAG3), a major receptor of pathologic α-syn transmission. Our preliminary results showed that heterochronic blood exchange (HBE) from young mice can inhibit pathologic α-syn transmission to cells and inflammation in aged mice. We further identified two blood-borne aging-modulated proteins that regulate LAG3-mediated pathologic α-syn transmission. The first plasma protein fibrinogen-like protein (FGL1) as the major inhibitory ligand of LAG3, is decreased by aging and inhibits α-syn transmission. The second plasma protein sLAG3 is the soluble form of LAG3 protein, and it is increased by aging and promotes α-syn transmission. Our studies also support the feasibility to use HBE approach to modulate plasma levels of FGL1 and sLAG3 in aged mice by young blood. Our Central Hypothesis is that HBE with young blood inhibits α-synucleinopathy through two aging-associated circulatory proteins (FGL1 and sLAG3) essential for LAG3-mediated pathologic α-syn transmission. FGL1 functions as a rejuvenation factor to inhibit pathologic α-syn spreading in the gut-brain axis and consequent neurodegeneration, neuroinflammation, and behavioral deficits associated α-synucleinopathy. sLAG3 acts as an age-acceleration factor with antagonistic functions to FGL1. In specific aim 1, we propose to determine if HBE ameliorates α-synucleinopathy and related cognitive impairment by increasing aging-reduced FGL1 to inhibit pathological α-syn spreading. In specific aim 2, we propose to determine if HBE ameliorates α-synucleinopathy and related cognitive impairment by decreasing aging-induced sLAG3 to inhibit pathological α-syn spreading. Modulating plasma factors is a novel strategy to inhibit pathologic α-syn spreading and treating α-synucleinopathy and dementia. Positive results from this study will justify the development of novel α-synucleinopathy therapies based on plasma factor modulation. Novel molecular insights from this project will lay a solid foundation for the optimization and clinical translation of α-synucleinopathy therapies based on FGL1 and sLAG3 modulation.

NIH Research Projects · FY 2025 · 2021-09

Alzheimer’s disease is a major threat to public health. Because Alzheimer’s disease has no cure, it is critical to identify its modifiable risk factors that can be targeted to reduce its burden. Although initial evidence suggests its plausibility, relatively little attention has been paid to the role of common infections in Alzheimer’s disease etiology. We propose to investigate the association of infection with common pathogens—Herpes Simplex Virus Types 1 and 2, Cytomegalovirus, Epstein-Barr Virus, Toxoplasma gondii—measured four times over ~25 years, and SARS-CoV-2 (the virus that causes COVID-19), with: (a) cognitive decline, and adjudicated mild cognitive impairment (MCI) and dementia diagnoses; (b) plasma biomarkers of Alzheimer’s disease; and (c) markers of physiological aging (telomere shortening, cyclin-dependent kinase inhibitor p16INK4a, plasma-derived senescence-associated secretory phenotypes, and epigenetic clocks). We will also explore sex, Alzheimer’s disease risk genes, and stress-related exposures (mental disorders and their symptoms, stressful life events, and poor sleep) as moderators that amplify the risk of adverse infection-induced cognitive, brain health, and physiological aging outcomes. Inclusion of viral specific CD8 T-cell differentiation in combination with antibody levels measured serially in the same individuals will allow us to distinguish between long-term infections and reactivation and to evaluate the influence of the course of both infection and immune response to infection, on our outcomes. Senescence-associated secretory phenotypes will point to novel senescent pathways by which infections affect brain health. We will accomplish this using existing data and collecting new data from participants in the Baltimore Epidemiological Catchment Area (ECA) Study Follow-up, which has been assessed five times for >35 years (mean age = 70 years, range 58-100). Blood specimens have been collected three times over ~25 years in the ECA, and we will collect an additional blood draw to obtain infection status at four time points, providing a rare opportunity to quantify timing of exposure and reactivation of latent infections in relation to cognitive and functional decline and Alzheimer’s disease biomarkers and potential pathways. The MPIs of the proposed study are currently completing Wave 5 of data collection in the ECA, including measures of cognitive and functional decline, adjudicated MCI and dementia diagnoses, cellular aging and genome-wide genetic and epigenetics assays. Our preliminary data in the ECA link common pathogens of interest with lower cognitive performance and suggest effect modification by apolipoprotein E genotype. Our team consists of experts in cognitive aging and Alzheimer’s disease, neurovirology, neuropsychology, Alzheimer’s disease biomarkers, genetics and epigenetics, and the biology of aging. Results will clarify the extent to which common infections increase the risk for Alzheimer’s disease and related dementias, and because this work is performed in a longitudinal cohort, it will elucidate mechanisms, identify moderators and candidate pathways which precede decline, thus informing future preventive, and perhaps therapeutic, efforts.

NIH Research Projects · FY 2025 · 2021-09

Neurodevelopmental processes are shaped by dynamic interactions between genes and environments. Maladaptive experiences early in life can alter developmental trajectories, leading to harmful and enduring developmental sequelae. Pre- and postnatal hazards include maternal substance exposure, toxicant exposures in pregnancy and early life, maternal health conditions, parental psychopathology, maltreatment, and excessive stress. To elucidate how various environmental hazards impact child development, it is imperative that a normative template of developmental trajectories over the first 10 years of life be established based on a sufficiently large and demographically heterogeneous sample of the US population. To accomplish this, the Healthy Brain and Child Development (HBCD) Consortium has been formed to deploy a harmonized, optimized, and innovative set of neuroimaging (MRI, EEG) measures complemented by an extensive battery of behavioral, physiological, and psychological tools, and biospecimens to understand neurodevelopmental trajectories in a sample of 7,200 mothers and infants enrolled at 27 sites across the United States (US). The HBCD Study will carry out a common research protocol under direction of the HBCD Consortium Administrative Core (HCAC) and will assemble and distribute a comprehensive and well-curated research dataset to the scientific community at large under the direction of the HBCD Data Coordinating Center (HDCC). The overarching goal of the HBCD Study is to create a comprehensive, harmonized, and high-dimensional dataset that will characterize typical neurodevelopmental trajectories in US children and that will assess how biological and environmental exposures affect those trajectories. A special emphasis will be placed on understanding the impact of pre- and postnatal exposure to opioids, marijuana, alcohol, tobacco and/or other substances. To address these broad objectives, the sample of women enrolled will include: 1) a varied cohort that is representative of the US population; 2) pregnant woman with use of targeted substances (opioids, marijuana, alcohol, tobacco); and 3) demographically and behaviorally similar women without substance use in pregnancy to enable valid causal inferences. In addition, the HBCD Study will identify key developmental windows during which both harmful and protective environments have the most influence on later neurodevelopmental outcomes. The large, multi-modal, longitudinal, and generalizable dataset that will be produced for the first time by this study will provide novel insights into child development using state-of-the-art methods. The HBCD Study will inform public policy to improve the health and development of children across the nation. 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.