Projects

ABSTRACT Single-cell genomic, epigenomic, and transcriptomic technologies can identify unique cell subsets with important physiologic roles; however, RNA or DNA signatures cannot always be linked to unique surface markers, hampering the re-isolation of these cell subsets for in-depth analyses. Moreover, conventional single-cell methods require sequencing prohibitively large numbers of cells to characterize rare subsets. Here we will develop and apply SEARCH-seq, a high-throughput cytometry method that detects RNA or DNA markers with single molecule sensitivity that allows the rapid isolation of target cells for in-depth transcriptomic, genomic, or epigenomic analyses. We will use the method to study the regulatory mechanisms controlling an astrocyte subpopulation characterized by an alternatively spliced XBP1 transcript, which promotes disease pathology in multiple sclerosis (MS). This subpopulation also manifests in the pre-clinical mouse model of experimental autoimmune encephalomyelitis (EAE). Using SEARCH-seq in combination with conditional knock-out mice, in vivo CRISPR/Cas9-driven perturbation studies, and RNA-seq analyses of mouse EAE and human MS samples, we will characterize the role of these cells and their interaction with the nuclear receptor NR3C2 and its corepressor NCOR2 in limiting XBP1-driven pathogenic astrocyte responses. In summary, SEARCH-seq is a novel, sensitive, and high throughput method to capture rare brain cell subsets that are difficult to study with existing technology and may have therapeutically targetable mechanisms relevant to MS pathogenesis.

ABSTRACT Infections by different pathogens can manifest with similar symptoms, but appropriate treatment requires specific and accurate diagnosis. Clinicians often turn to multiplexed assays testing for many organisms (e.g. BioFire). While these approaches can test for 50-70 organisms, they do not provide concentration titers, which is necessary to identify the causative pathogen among the several false positives or clinically meaningless commensals. As a result, the clinician must perform additional tests to identify which of the positives is causative. Although these tests use quantitative PCR, in clinical labs the results are reported as presence/absence due to the finicky nature of PCR in this setting, which is sensitive to minor variations in reaction efficiency, operator variability. As a result, today, only a few widespread PCR tests are FDA approved to report quantitative result. In contrast to qPCR, digital PCR (dPCR) measures target titers by counting individual molecules. As a result, dPCR provides an absolute concentration measurement that doesn’t require a standard curve. In addition, the reaction is cycled to endpoint, then quantified; it does not require careful estimation of the amplification rate, which is a major source of variability in qPCR. Thus, dPCR is less sensitive to variations in reaction efficiency and provides superior consistency. However, current dPCR methods are limited in multiplexing, allowing just 5- 6 targets per assay, while qPCR can test up to 100. Moreover, dPCR requires complex microfluidic equipment that burdens testing lab personnel and increases cost. Until these issues can be addressed, qPCR will continue to dominate the clinical lab, and quantitative and absolute pathogen load reporting will remain beyond reach. Here, we propose a novel nucleic acid technology combining the quantitativeness and robustness of dPCR with the simplicity and multiplexing of qPCR. Our vision is to enable broad spectrum detection wherein each pathogen is associated with a high confidence, quantitative titer. Our approach – gigapixel PCR (gPCR) – is enabled by our recent discoveries of self-assembled partitioning, for microfluidic-free generation of monodispersed emulsions, and linearized target quantitation with capillary electrophoresis (CE). CE allows sensitive quantitation over 7 decades and provides amplicon length information with single nucleotide resolution. In gPCR, we use this to perform multiplexed detection of over 100 amplicons per reaction. In contrast to qPCR, which requires that the sample be split to test for different targets, thereby diluting it and reducing sensitivity, with gPCR the targets are tested without splitting, maintaining them at maximal concentration, and substantially increasing sensitivity. Moreover, based on robust dPCR, gPCR provides reproducible, quantitative results across testing conditions. It thus addresses the major limitations of current dPCR technologies and provides the first viable alternative to qPCR in the clinic. We will develop and validate the technology against accepted standards (SeraCare), and work with our longstanding collaborators (Drs. Melanie Ott and Charles Chiu) to apply it to respiratory and CNS infections from samples previously collected at UCSF hospitals.

ABSTRACT Infectious diseases remain a significant cause of morbidity worldwide, highlighting the critical need for accurate diagnosis. However, shared symptoms among different infections and the emergence of drug resistance make diagnosis and treatment selection challenging. In this competing renewal, we propose to develop Gigapixel NGS (gNGS) to enable rapid, sensitive, and information-rich infectious disease diagnosis. gNGS builds upon our Gigapixel PCR (gPCR) technology by incorporating powerful next-generation sequencing capabilities. By utilizing double emulsion vesicles for single cell assays, gNGS eliminates the need for specialized droplet analyzers and allows common flow cytometers to be used for genome isolation. Our goal in this renewal is to leverage the capabilities of Gigapixel NGS to detect, isolate, and sequence infectious pathogen genomes from patient samples, which will improve the efficiency and sensitivity of pathogen sequencing. We will collaborate with Dr. Charles Chiu, a renowned expert in infectious disease diagnostics who leads the CLIA-certified pathogen lab at UCSF, to develop clinical workflows and bioinformatic tools for interrogating the recovered genomes for relevant biomarker sequences, including virulence factors and drug resistance genes. Dr. Chiu's expertise in infectious disease diagnostics and practical experience in clinical sample sequencing for pathogen detection will ensure that the new diagnostic is effective and practical in a clinical setting.

ABSTRACT Infections by different pathogens can manifest with similar symptoms, but appropriate treatment requires specific and accurate diagnosis. Clinicians often turn to multiplexed assays testing for many organisms (e.g. BioFire). While these approaches can test for 50-70 organisms, they do not provide concentration titers, which is necessary to identify the causative pathogen among the several false positives or clinically meaningless commensals. As a result, the clinician must perform additional tests to identify which of the positives is causative. Although these tests use quantitative PCR, in clinical labs the results are reported as presence/absence due to the finicky nature of PCR in this setting, which is sensitive to minor variations in reaction efficiency, operator variability. As a result, today, only a few widespread PCR tests are FDA approved to report quantitative result. In contrast to qPCR, digital PCR (dPCR) measures target titers by counting individual molecules. As a result, dPCR provides an absolute concentration measurement that doesn’t require a standard curve. In addition, the reaction is cycled to endpoint, then quantified; it does not require careful estimation of the amplification rate, which is a major source of variability in qPCR. Thus, dPCR is less sensitive to variations in reaction efficiency and provides superior consistency. However, current dPCR methods are limited in multiplexing, allowing just 5- 6 targets per assay, while qPCR can test up to 100. Moreover, dPCR requires complex microfluidic equipment that burdens testing lab personnel and increases cost. Until these issues can be addressed, qPCR will continue to dominate the clinical lab, and quantitative and absolute pathogen load reporting will remain beyond reach. Here, we propose a novel nucleic acid technology combining the quantitativeness and robustness of dPCR with the simplicity and multiplexing of qPCR. Our vision is to enable broad spectrum detection wherein each pathogen is associated with a high confidence, quantitative titer. Our approach – gigapixel PCR (gPCR) – is enabled by our recent discoveries of self-assembled partitioning, for microfluidic-free generation of monodispersed emulsions, and linearized target quantitation with capillary electrophoresis (CE). CE allows sensitive quantitation over 7 decades and provides amplicon length information with single nucleotide resolution. In gPCR, we use this to perform multiplexed detection of over 100 amplicons per reaction. In contrast to qPCR, which requires that the sample be split to test for different targets, thereby diluting it and reducing sensitivity, with gPCR the targets are tested without splitting, maintaining them at maximal concentration, and substantially increasing sensitivity. Moreover, based on robust dPCR, gPCR provides reproducible, quantitative results across testing conditions. It thus addresses the major limitations of current dPCR technologies and provides the first viable alternative to qPCR in the clinic. We will develop and validate the technology against accepted standards (SeraCare), and work with our longstanding collaborators (Drs. Melanie Ott and Charles Chiu) to apply it to respiratory and CNS infections from samples previously collected at UCSF hospitals.

ABSTRACT Single-cell genomic, epigenomic, and transcriptomic technologies can identify unique cell subsets with important physiologic roles; however, RNA or DNA signatures cannot always be linked to unique surface markers, hampering the re-isolation of these cell subsets for in-depth analyses. Moreover, conventional single-cell methods require sequencing prohibitively large numbers of cells to characterize rare subsets. Here we will develop and apply SEARCH-seq, a high-throughput cytometry method that detects RNA or DNA markers with single molecule sensitivity that allows the rapid isolation of target cells for in-depth transcriptomic, genomic, or epigenomic analyses. We will use the method to study the regulatory mechanisms controlling an astrocyte subpopulation characterized by an alternatively spliced XBP1 transcript, which promotes disease pathology in multiple sclerosis (MS). This subpopulation also manifests in the pre-clinical mouse model of experimental autoimmune encephalomyelitis (EAE). Using SEARCH-seq in combination with conditional knock-out mice, in vivo CRISPR/Cas9-driven perturbation studies, and RNA-seq analyses of mouse EAE and human MS samples, we will characterize the role of these cells and their interaction with the nuclear receptor NR3C2 and its corepressor NCOR2 in limiting XBP1-driven pathogenic astrocyte responses. In summary, SEARCH-seq is a novel, sensitive, and high throughput method to capture rare brain cell subsets that are difficult to study with existing technology and may have therapeutically targetable mechanisms relevant to MS pathogenesis.

Project Summary / Abstract Dr. Mahsa Abassi is an Assistant Professor of Medicine in the Division of Infectious Diseases at the University of Minnesota. Over the past six years, she has been engaged in clinical research, focusing on HIV- related neuroinfections in Uganda. Her long-term objective is to become an independent clinical researcher with an emphasis on improving outcomes in neuroinfections. Her career development plan proposes mentored training in: 1) neurologic techniques (EEGs, neuroradiology, and neurocognitive assessment), 2) laboratory techniques related to metabolomics applications, and 4) biostatistics with an emphasis of analyzing metabolites in biologic samples. Research: Cryptococcal meningitis accounts for 15% of HIV/AIDS-related deaths globally and is the most common cause of adult meningitis in Africa. Altered mental status (ranging from delirium to coma) at the time of cryptococcal meningitis diagnosis is consistently an independent predictor of increased mortality. Despite repeated studies confirming this strong association between altered mental status and death, there is a fundamental lack of understanding into the exact neurological abnormalities leading to acute altered mental status, its contributions to increased mortality, and the best practices for management. The objective of this proposal is to identify the neurological abnormalities that contribute to altered mental status and to understand how this contributes to increased cryptococcal mortality. The overarching hypothesis is that cryptococcal meningitis with its increased intracranial pressure leads to cerebral hypoxia, abnormal electrical activity, and biochemical changes in the central nervous system (CNS) that can be detected through brain metabolite CSF analysis and enhanced clinical monitoring with cerebral oximetry and EEGs. This proposal aims to: 1) determine if HIV-infected persons with cryptococcal meningitis presenting with altered mental status (Glasgow Coma Scale (GCS) <15) at diagnosis have measurable underlying neurological abnormalities and impairments in cerebral energy metabolism (i.e. insufficient oxidative metabolism) as compared to persons with normal mental status (GCS=15); and 2) determine if implementation of standardized clinical interventions can reverse neurological abnormalities and improve cerebral energy metabolism within 3 days of diagnosis, and reduce 30-day mortality in HIV-infected persons with cryptococcal meningitis presenting with altered mental status (GCS<15). Results of the above aims will shed light into previously unknown pathophysiologic mechanisms that lead to altered mental status in cryptococcal meningitis. The training in neuroinfections, metabolomics applications, and biostatistics that Dr. Abassi will obtain will inform future proposals dedicated to understanding the neuropathology of various neuroinfections and finding evidence- based interventions dedicated to improving survival.

Dr. Mahsa Abassi is an Assistant Professor of Medicine in the Division of Infectious Diseases at the University of Minnesota. She has been engaged in clinical research, focusing on AIDS related neuroinfections in Uganda. Her long-term objective is to become an independent clinical researcher with an emphasis on improving outcomes in neuroinfections. Dr. Abassi’s Career Development Plan will provide her with the mentorship support, didactic course work, and hands-on training she will need to become an expert in clinical and translational research with a focus on HIV-associated neuroinfections. Her career development plan proposes mentored training in: 1) neurologic techniques (EEGs and neuroradiology), 2) laboratory techniques related to metabolomics applications, 3) advanced training in clinical research and translational medicine, and 4) biostatistics with an emphasis of analyzing metabolites in biologic samples. Research: Cryptococcal meningitis accounts for 15% of HIV/AIDS-related deaths, globally. Altered mental status at the time of cryptococcal meningitis diagnosis is consistently an independent predictor of increased mortality. Despite repeated studies confirming this strong association between altered mental status and death, there is a fundamental lack of understanding into the exact neurological abnormalities leading to altered mental status, its contributions to increased mortality, and best practices for management. The objective of this proposal is to identify the neurological abnormalities that contribute to altered mental status and to understand how it contributes to increased cryptococcal mortality. The overarching hypothesis is that cryptococcal meningitis with its increased intracranial pressure leads to cerebral hypoxia, abnormal electrical activity, and biochemical changes in the central nervous system (CNS) that can be detected through brain metabolite CSF analysis and enhanced clinical monitoring with cerebral oximetry, EEGs, and ECGs. This proposal aims to 1) determine if HIV-infected persons with cryptococcal meningitis presenting with altered mental status (Glasgow Coma Scale (GCS) <15) at diagnosis have measurable underlying neurological abnormalities and impairments in cerebral energy metabolism (i.e. insufficient oxidative metabolism) as compared to persons with normal mental status (GCS=15), and 2) determine if implementation of standardized clinical interventions can reverse neurological abnormalities and improve cerebral energy metabolism within 3 days of diagnosis,, and reduce 30-day mortality in HIV-infected persons with cryptococcal meningitis and altered mental status (GCS<15). Results of the above aims will shed light into previously unknown pathophysiologic mechanisms that lead to altered mental status in cryptococcal meningitis. The training in metabolomics applications and clinical research that Dr. Abassi will obtain will inform future proposals on evidence-based interventions dedicated to improving survival in HIV-associated neuroinfections.

Project Summary/Abstract Acute exacerbations of chronic obstructive pulmonary disease (COPD) result in significant morbidity and healthcare costs, especially severe exacerbations that require hospitalization. Traditional bronchodilators and anti- inflammatory medications, as well as programs to monitor and treat symptoms, have a modest effect on reducing hospital admissions. Readmission rates remain unacceptably high, so alternative approaches are needed. In this regard, pulmonary rehabilitation (PR) is remarkably effective, and is associated with an 80% reduction in admission rates. Successful completion of PR is also associated with substantial improvements in quality of life, dyspnea, as well as exercise tolerance. Despite these benefits, recent studies have highlighted very poor referral rates for PR overall and as low as 1.9% post-hospitalization, and non-completion rates as high as 60%. The poor adaptation of PR in the community is due to a combination of barriers in availability and accessibility, as well as attrition. The number of PR centers in the United States is inadequate, and these centers are mostly distributed in urban areas. In addition, multiple socioeconomic and medical barriers hinder access to PR, and contribute to high drop-out rates. New strategies are needed to reduce hospital readmission in COPD, and to increase the delivery of PR to underserved urban as well as rural areas in the community. Given the scarcities in existing resources, conventional models of care delivery are being challenged, and attempts are being made to find alternative, cost-effective ways of delivering healthcare to a larger number of eligible patients. To overcome the socioeconomic and physical barriers to PR, we hypothesize that a video telehealth intervention that will deliver PR to the patient's home, regardless of geographic location, will reduce hospital readmissions in COPD, and reduce respiratory morbidity. To test our hypothesis we propose a prospective randomized controlled phase 3 multicenter clinical trial comparing a real-time video telehealth PR intervention plus standard of care versus standard of care alone, with the following Specific Aims: (1) To determine if a video telehealth PR intervention reduces 30-day all-cause readmissions in patients hospitalized for acute exacerbation of COPD, (2) To evaluate the effects of the video telehealth PR intervention on dyspnea and respiratory quality of life in COPD post hospital discharge, and (3) To evaluate the cost-effectiveness of the telehealth intervention. Accomplishment of the aims of this study will result in a significant reduction in COPD readmission rates, and a paradigm shift in the way PR is delivered to patients with COPD, especially those that reside in remote and rural areas with limited access to pulmonary rehabilitation.

OTHER PROJECT INFORMATION – Project Summary/Abstract Does Health Insurance Promote Health through Providers and Networks? Through their varying provider networks and coverages, health insurance plans directly and indirectly steer patients toward some clinicians and hospitals and away from others. If providers differ in their quality of care and especially if they provide better care for certain types of patients, such network variation can meaningfully affect health outcomes at the plan level, and not just the provider level. Yet, consumers selecting health plans lack information about which plans will direct them to better providers, obscuring this potentially important channel to improving population health. This project will develop novel outcome-based measures of insurance plan quality based on where enrollees receive care, and taking account how causal impacts on health vary across provider networks. By furthering the scientific understanding of the relationship between provider quality and patient outcomes at the plan level, we aim to equip patients with new information for selecting insurance plans. Our measures will also provide regulators with new instruments to reward plans that make people healthier. At the core of our research plan are new, causal estimates of how mortality impacts vary across hospitals and physicians and, in turn, across the health plans that rely on different provider networks. We focus on older Americans enrolled in Medicare Advantage plans, leveraging novel data on the provider networks used by different Medicare Advantage plans. The estimates will characterize which hospitals and providers produce the best outcomes for beneficiaries with different demographics and comorbidities and thus which plans are suitable for each type of patient by directing them to providers who produce good outcomes for conditions they have or are likely to develop. The project has four primary aims: Aim 1: Measure and Validate Mortality Effects for Healthcare Providers Aim 2: Link Physician and Hospital Mortality Effects to Plan Mortality Effects Aim 3: Explore Heterogeneity in Mortality Effects Across Beneficiaries Aim 4: Evaluate the Impact of Policy Proposals

Increasing evidence suggests that the intestine plays an important role in sensing not only the presence of pathogens but also changes in the microbiota, which ultimately result in changes in the regulation of immune pathways and behaviors by communicating with neurons. However, the complexity of the nervous and immune systems of mammals makes it difficult to dissect the mechanisms by which the neural-gut axis communicates using bidirectional signals to control intestinal immunity. Studies in the nematode Caenorhabditis elegans show that bacterial colonization of the intestine results in the activation of the expression of innate immune genes in the gut and the activation of a neuroendocrine signal that controls pathogen avoidance. The germline also plays a key role in the neural-gut axis not only by transmitting the immunological memory to the next generation but also by communicating pathogenic cues that travel from the gut to the nervous system to control innate immunity. The long-term goal of this proposal is to elucidate the mechanism by which the neural-germline-gut axis communicates to sense pathogens and/or infection-induced physiological changes to control innate immunity at the whole animal level. Thus, we will explore the general hypothesis that the neural-germline-gut axis plays a critical role in the organismal response against bacterial pathogens by helping the nervous system integrate signals from infected sites and different tissues to coordinate the immune response. Specific genes and neurons will be studied to dissect the neural circuits that regulate immune activation in response to pathogen exposure and pathogen-induced alterations of the animal’s physiology. A genetics approach will also be used to identify neurotransmitters and endocrine signals potentially involved in the neural-immune communication that takes place between neurons and different tissues and infected sites. RELEVANCE (See instructions): The systemic control of innate immunity is critical because inflammation accounts for the major physiological, metabolic, and pathological responses to infections. We plan to continue our studies to clarify the role of the nervous system in the regulation of intestinal innate immune responses against bacterial pathogens. A better understanding of the neural-immune communication could lead to new therapeutic targets for diseases involving a deficient innate immune system.

Increasing evidence suggests that the intestine plays an important role in sensing not only the presence of pathogens but also changes in the microbiota, which ultimately result in changes in the regulation of immune pathways and behaviors by communicating with neurons. However, the complexity of the nervous and immune systems of mammals makes it difficult to dissect the mechanisms by which the neural-gut axis communicates using bidirectional signals to control intestinal immunity. Studies in the nematode Caenorhabditis elegans show that bacterial colonization of the intestine results in the activation of the expression of innate immune genes in the gut and the activation of a neuroendocrine signal that controls pathogen avoidance. The germline also plays a key role in the neural-gut axis not only by transmitting the immunological memory to the next generation but also by communicating pathogenic cues that travel from the gut to the nervous system to control innate immunity. The long-term goal of this proposal is to elucidate the mechanism by which the neural-germline-gut axis communicates to sense pathogens and/or infection-induced physiological changes to control innate immunity at the whole animal level. Thus, we will explore the general hypothesis that the neural-germline-gut axis plays a critical role in the organismal response against bacterial pathogens by helping the nervous system integrate signals from infected sites and different tissues to coordinate the immune response. Specific genes and neurons will be studied to dissect the neural circuits that regulate immune activation in response to pathogen exposure and pathogen-induced alterations of the animal’s physiology. A genetics approach will also be used to identify neurotransmitters and endocrine signals potentially involved in the neural-immune communication that takes place between neurons and different tissues and infected sites. RELEVANCE (See instructions): The systemic control of innate immunity is critical because inflammation accounts for the major physiological, metabolic, and pathological responses to infections. We plan to continue our studies to clarify the role of the nervous system in the regulation of intestinal innate immune responses against bacterial pathogens. A better understanding of the neural-immune communication could lead to new therapeutic targets for diseases involving a deficient innate immune system.

PROJECT SUMMARY/ABSTRACT Nearly half of the ~120 million Americans with hypertension are salt-sensitive and confer a higher risk of cardiovascular and kidney disease. Only 26% of hypertensive patients reach blood pressure (BP) control, indicating the urgent need to study novel mechanistic pathways in hypertension. The gut microbiota and immune system are critical in hypertension in males; this is largely unknown in females. This grant will address this knowledge gap by testing the role of the gut microbiota and its derived metabolites to modulate immune mechanisms known to contribute to salt-sensitive hypertension and renal end-organ damage. Mechanistic studies will be performed in male and female Dahl Salt-Sensitive (SS) rats, with translational studies performed in stored biological samples from humans consuming a diet high in salt. Dahl SS rats, a model consistent with human salt-sensitive hypertension, exhibit a greater degree of salt- sensitivity and associated renal end-organ damage in males compared to females. Through microbiota transfer studies, we know the gut microbiota plays a causal role in the regulation of BP and the extent of renal damage and renal T cell infiltration. We have observed stark sex differences in gut microbiota composition and gut- derived metabolites. It therefore begs the question of how sex-specific microbiota and its metabolites impact T cell function and ultimately, salt-sensitivity. This proposal, deemed meritorious for prior R56 support, will test the central hypothesis that the gut microbiota produces pathological metabolites in males (carnitine/TMAO) and protective metabolites in females (SCFAs), and these metabolites exert their actions on blood pressure and the kidney by altering T cell activation. Aim 1 will test whether sex differences in gut microbiota function alters the severity of salt-sensitive hypertension and renal damage. Sex-specific microbiota transfer and hormone manipulation studies (i.e. gonadectomy, Sry transgenic SS rat) will reveal whether the gut microbiota sex- and hormone-dependently contributes to salt- sensitivity in SS rats. Aim 2 will test whether the sex-specific influence of the gut microbiota on disease severity is mediated by gut-derived metabolites (increased TMAO in males versus increased SCFAs in females) and will determine the effect of these metabolites on T cell activation. Functional T cell changes in response to either sex-specific gut microbiota manipulation or gut metabolites will be assessed by measuring alterations to T cell gene expression, bioenergetics, and cytokine production. Parallel translational studies in urine, stool, serum, and PBMCs from humans consuming a diet high in salt will assess whether sex differences in microbiota, metabolites, and inflammation can explain sex differences in BP in humans. These studies will reveal key mechanisms related to gut-immune-kidney crosstalk and provide the preclinical and human basis for developing novel targets to improve BP control and kidney disease.

The U.S. population is aging rapidly, with a rising burden of chronic diseases, functional impairments, and cognitive decline among older adults. In parallel, major scientific advances have expanded our understanding of the biological mechanisms underlying aging and have introduced engineering and bioengineering innovations with the potential to extend healthspan, the period of life spent in good health. However, a persistent gap remains in translating these discoveries into real-world interventions that improve clinical outcomes and enhance healthspan for older adults. To address this translational gap, the Translational Aging Research Training Program was launched in 2018 to develop future leaders in aging research. Despite disruptions due to COVID-19, the program successfully trained 14 fellows from clinical, biological, and engineering, disciplines. This renewal proposes an enhanced program embedded in the interdisciplinary ecosystem of the Johns Hopkins Human Aging Project (HAP), an institutional initiative that foster collaborative research and training to extend the healthspan and improve the lives of older adults. This program will continue to leverage strong scientific leadership, a rich research infrastructure, and a supportive environment that welcomes individuals from all backgrounds committed to excellence in aging research. Its specific aims are to: 1) Deliver an interdisciplinary training experience in translational aging research for postdoctoral fellows (MD and/or PhD), structured around core coursework, interdisciplinary meetings, weekly translational aging seminars, individualized development plans, and leadership training. 2) Provide each fellow with a mentored research experience in translational aging science, either in biology, engineering, or clinical research. 3) Enable access to a broad network of resources and aging-focused research infrastructure at Johns Hopkins. 4) Prepare trainees for long-term success in academic careers through individualized mentoring and professional development activities. By the end of the program, trainees will possess: -Interdisciplinary expertise in aging biology, clinical geriatrics, and engineering approaches to aging-related problems; -Skills in translational science to move discoveries from bench to bedside; -Competencies in leadership and collaboration to pursue impactful research careers that aim to extend healthspan and improve aging-related outcomes.

Resource Core 2 (RC2) Biological Mechanisms Core: Project Summary The identification of the etiologies of frailty and age-related vulnerability remains a crucial challenge for gerontological research. Key to this challenge are the development of a better understanding of the underlying biological basis that contributes to frailty and the identification of key biological pathways for the development of interventions that might help prevent or alleviate frailty and loss of independence. The goal of Johns Hopkins Older Americans Independence Center (OAIC) Biological Mechanisms Core (RC-2) is to enable the next generation of frailty-related etiological discovery and to promote the translation of these discoveries into clinically relevant diagnostic, preventive, and treatment modalities. This will be achieved through the provision of high-quality biological and bioengineering measurement expertise, incorporation of new technologies, analytical and computational expertise for genetics and omics analyses, and infrastructure necessary to attain this goal. In order to comprehensively encompass the biological expertise necessary to study frailty-related etiology, we have engaged a leadership team and internal consultants with complementary and synergistic biological and translational expertise needed to unravel the complex biological mechanisms that underpin frailty. They also all bring mentorship skills for trainees, and infrastructure to RC-2 and national prominence to frailty research. The specific aims of RC-2 are to: 1) provide state of the art scientific expertise, infrastructure, and technology necessary to advance biological and etiological research related to frailty, 2) provide access to biological samples from human subjects and from animal models necessary to test hypotheses related to frailty, 3) facilitate the translation of biological findings into interventions or prevention-focused clinical studies, 4) provide training, mentorship, and guidance to promising junior investigators around biological mechanisms that impact frailty, and 5) provide institutional and external visibility for RC-2 related science and activities. Our aims will be accomplished through close communication between the core leaders and their laboratories, close partnership with the other OAIC cores, and the engagement of expert consultants in the highly relevant areas of mitochondrial measurement, metabolomics, epigenetics, mouse model development, nanotechnologies for diagnostic and treatment development, and the development of multi-omic analyses related to frailty. By providing these resources, RC-2 will foster high quality research that elucidates clinically relevant biological pathways that underlie frailty and related interventions that hold promise to attenuate frailty, related conditions, and the loss of independence.

Dr. Peter Abadir is an active Geriatrician and associate professor of Medicine, Electrical, and Computer Engineering at Johns Hopkins University (JHU). He has put forth this K24 Mid-Career Development proposal dedicated to mentorship in patient-centric translational research. Dr. Abadir’s research connects molecular changes associated with aging to physical and cognitive declines observed in older adults. He's been instrumental in bridging the fields of aging research and engineering at JHU, leading to the creation of both the Hopkins GeroTech Program and Artificial Intelligence and Technology Collaboratory (AITC) for Aging Research. Candidate: Dr. Abadir is committed to further training to broaden his research program and mentor emerging scholars from both biology and engineering. Recognized as an accomplished clinician-scientist, he boasts significant achievements in translational research, particularly concerning frailty and Alzheimer's disease. His escalating leadership roles span both institutional and national platforms centered on patient-oriented research and aging-focused mentorship. As the co-PI of the Johns Hopkins AITC and the co-director of its Clinical Translation and Validation Core, Dr. Abadir actively contributes to the innovation of technologies tailored for older adults. Furthermore, he directs the molecular measurement core at the Older American Independence Center (OAIC) and is the associate director of the Translational Aging Research Training Program (T32). Mentoring Plan/Environment: Dr. Abadir's K24 proposal taps into the wealth of training assets at Johns Hopkins including resources from the JHU AITC, OAIC, the Institute for Clinical and Translational Research, and various T32 training grants. Special attention is given to mentoring investigators focused on populations at highest risk for aging-related conditions. Ideal mentees exhibit interest at the intersection of aging, technology, and Geroscience. The structured mentoring approach categorizes mentees based on experience, allocating specific effort percentages to ensure quality interactions. Regular individual meetings, hands-on research training, data interpretation, and presentation skills enhancement form core of mentoring strategy. Multi-tiered evaluations ensure consistent mentorship quality. This strategy aims to cultivate future leaders in translational aging research from both biological and engineering fields. Research Plan: The pioneering research funded by this K24 award aims to harness Artificial Intelligence (AI) tools and analytics for assessing physical and cognitive digital biosignals in frail older individuals including those with Alzheimer’s disease and associating these with molecular markers. The study will evaluate the accuracy and reliability of a new method that uses AI to analyze non-invasive multimodal biometric signals like speech, voice, eye movements, handwriting, and gait. This methodology will enhance our understanding of the varied characteristics of frailty in older individuals and pinpoint early cognitive shifts, including signs of Alzheimer’s. Building upon the existing efforts at Johns Hopkins AITC and OAIC, this research seeks to uncover new insights and broaden avenues for upcoming researchers.

Resource Core 2 (RC2) Biological Mechanisms Core: Project Summary The identification of the etiologies of frailty and age-related vulnerability remains a crucial challenge for gerontological research. Key to this challenge are the development of a better understanding of the underlying biological basis that contributes to frailty and the identification of key biological pathways for the development of interventions that might help prevent or alleviate frailty and loss of independence. The goal of Johns Hopkins Older Americans Independence Center (OAIC) Biological Mechanisms Core (RC-2) is to enable the next generation of frailty-related etiological discovery and to promote the translation of these discoveries into clinically relevant diagnostic, preventive, and treatment modalities. This will be achieved through the provision of high-quality biological and bioengineering measurement expertise, incorporation of new technologies, analytical and computational expertise for genetics and omics analyses, and infrastructure necessary to attain this goal. In order to comprehensively encompass the biological expertise necessary to study frailty-related etiology, we have engaged a leadership team and internal consultants with complementary and synergistic biological and translational expertise needed to unravel the complex biological mechanisms that underpin frailty. They also all bring mentorship skills for trainees, and infrastructure to RC-2 and national prominence to frailty research. The specific aims of RC-2 are to: 1) provide state of the art scientific expertise, infrastructure, and technology necessary to advance biological and etiological research related to frailty, 2) provide access to biological samples from human subjects and from animal models necessary to test hypotheses related to frailty, 3) facilitate the translation of biological findings into interventions or prevention-focused clinical studies, 4) provide training, mentorship, and guidance to promising junior investigators around biological mechanisms that impact frailty, and 5) provide institutional and external visibility for RC-2 related science and activities. Our aims will be accomplished through close communication between the core leaders and their laboratories, close partnership with the other OAIC cores, and the engagement of expert consultants in the highly relevant areas of mitochondrial measurement, metabolomics, epigenetics, mouse model development, nanotechnologies for diagnostic and treatment development, and the development of multi-omic analyses related to frailty. By providing these resources, RC-2 will foster high quality research that elucidates clinically relevant biological pathways that underlie frailty and related interventions that hold promise to attenuate frailty, related conditions, and the loss of independence.

PROJECT SUMMARY/ABSTRACT Resilience, defined as the ability to respond to stressors, declines with age and co-morbid conditions in older organisms. To date, little progress has been made to improve our understanding of resilience with aging and its multi-component dimensions in response to physical, cognitive, and social stressors, in part because resilience itself is likely a dynamic multi-systemic process that is most evident under conditions of stress. This application is for a five-year cooperative conference grant to continue the successful “Geriatrics Research: Bench-to-Bedside” series. The series has had an outstanding impact. Past conferences represented “paradigm-shifting” moments and “pivotal” junctures in areas such as frailty, multimorbidity, and delirium. Here we propose to build bridges and bring together resilience scientists and experts from the physical, cognitive, and social domains to discuss, debate, and learn from each other the dynamic multi-component phenomena of resilience. In the first conference, Overview of the Resilience World: State of the Science, we will focus on setting the stage and discuss working definitions of resilience from physical, cognitive, and psychosocial fields. A goal will be to identify areas of commonalities and differences and how we can operationalize a shared concept so that “resilience” can be quantified appropriately in different contexts. In the second conference, Stress tests and Biomarkers of Resilience, we aim to discuss resilience stress test paradigms and molecular markers that may be associated with resilient outcomes. Building on the first conference, this conference will refer to operationalized definitions of resilience and resilient outcomes, acknowledging that definitions may differ slightly across fields and contexts. Lastly, the third conference is entitled Optimizing Resilience. By integrating the foundation built in the first two conferences, we will focus on interventions to optimize resilience in older adults. This conference series, once completed, will serve as a forum and a unifying thread that will bring together national research leaders to build an integrative multidimensional understanding of resilience in older organisms.

ABSTRACT The Administrative Core (AC) will serve as the organizational hub of the Johns Hopkins Artificial Intelligence and Technology Collaboratory (JHU AITC), whose purpose is to advance artificial intelligence (AI) and technology solutions for aging and Alzheimer’s disease/Alzheimer’s disease–related dementias (AD/ADRD). Building on five years of successful operations (54 pilot awards, 7 commercialized products, 4 patents or patents-pending, and $11.7 million in external funding) the AC will provide governance, leadership, infrastructure, and accountability to ensure that the Collaboratory’s goals are met. The AC is co-directed by Phillip Phan, PhD, and Peter Abadir, MD, with oversight by the Executive Committee (EC, comprising the Multiple Principal Investigators), guidance from a Scientific Advisory Board (SAB), and day-to-day management by an experienced Executive Director. Specific Aim 1 is to guide the JHU AITC’s vision through interdisciplinary leadership and governance. The AC will coordinate scientific direction, cross-Core integration, fiscal oversight, and transparent decision-making, en- suring accountability to the EC, and SAB. Specific Aim 2 is to establish data standards, resource sharing, and dissemination. Collaborating with the Data Integrity & Computational Methods Core, the AC will implement FAIR (Findable, Accessible, Interoperable, Reusable) principles, create protocols for secure data access, and maintain a public website with training resources, solicitation announcements, and contributions to the Artificial Intelli- gence Technology Coordinating Center (AITCC)’s Living Textbook. Specific Aim 3 is to administer an annual national competition to identify and support innovative AI projects. Collaborating with the Research Innovation Core, the AC will oversee a rigorous two-stage peer review and ethics evaluation, aligned with the NIST AI Risk Management and SPIRIT-AI (Standard Protocol Items: Recommendations for Interventional Trials–Artificial In- telligence) checklists, to ensure scientific and clinical rigor and commercial viability. Specific Aim 4 is to collabo- rate with the AITCC on stakeholder engagement in ethical AI development. The AC will ensure that projects integrate perspectives of older adults, caregivers, clinicians, and industry, emphasizing fairness, privacy, and accessibility, particularly for rural and underserved populations. Specific Aim 5 is to closely partner with the AITCC in communication, reporting, and dissemination. Activities include quarterly and annual reporting, national symposia, publications, and broad engagement with academic, clinical, industry, and community stakeholders. The AC will apply a Donabedian structure–process–outcome framework to ensure reproducibility, accountability, and continuous improvement. Evaluation includes quarterly monitoring, fiscal oversight, benchmarking, and rapid responsiveness to awardees and stakeholders. By ensuring strong integration across Cores, accountability to the NIH/NIA mission, and broad dissemination of resources, the AC will advance a robust portfolio of AI inno- vations. Expected outcomes include enhanced reproducibility, accelerated translation of research into practice, commercial viability, and access to technology-enabled solutions for older adults.

The U.S. population is aging rapidly, with a rising burden of chronic diseases, functional impairments, and cognitive decline among older adults. In parallel, major scientific advances have expanded our understanding of the biological mechanisms underlying aging and have introduced engineering and bioengineering innovations with the potential to extend healthspan, the period of life spent in good health. However, a persistent gap remains in translating these discoveries into real-world interventions that improve clinical outcomes and enhance healthspan for older adults. To address this translational gap, the Translational Aging Research Training Program was launched in 2018 to develop future leaders in aging research. Despite disruptions due to COVID-19, the program successfully trained 14 fellows from clinical, biological, and engineering, disciplines. This renewal proposes an enhanced program embedded in the interdisciplinary ecosystem of the Johns Hopkins Human Aging Project (HAP), an institutional initiative that foster collaborative research and training to extend the healthspan and improve the lives of older adults. This program will continue to leverage strong scientific leadership, a rich research infrastructure, and a supportive environment that welcomes individuals from all backgrounds committed to excellence in aging research. Its specific aims are to: 1) Deliver an interdisciplinary training experience in translational aging research for postdoctoral fellows (MD and/or PhD), structured around core coursework, interdisciplinary meetings, weekly translational aging seminars, individualized development plans, and leadership training. 2) Provide each fellow with a mentored research experience in translational aging science, either in biology, engineering, or clinical research. 3) Enable access to a broad network of resources and aging-focused research infrastructure at Johns Hopkins. 4) Prepare trainees for long-term success in academic careers through individualized mentoring and professional development activities. By the end of the program, trainees will possess: -Interdisciplinary expertise in aging biology, clinical geriatrics, and engineering approaches to aging-related problems; -Skills in translational science to move discoveries from bench to bedside; -Competencies in leadership and collaboration to pursue impactful research careers that aim to extend healthspan and improve aging-related outcomes.

Dr. Peter Abadir is an active Geriatrician and associate professor of Medicine, Electrical, and Computer Engineering at Johns Hopkins University (JHU). He has put forth this K24 Mid-Career Development proposal dedicated to mentorship in patient-centric translational research. Dr. Abadir’s research connects molecular changes associated with aging to physical and cognitive declines observed in older adults. He's been instrumental in bridging the fields of aging research and engineering at JHU, leading to the creation of both the Hopkins GeroTech Program and Artificial Intelligence and Technology Collaboratory (AITC) for Aging Research. Candidate: Dr. Abadir is committed to further training to broaden his research program and mentor emerging scholars from both biology and engineering. Recognized as an accomplished clinician-scientist, he boasts significant achievements in translational research, particularly concerning frailty and Alzheimer's disease. His escalating leadership roles span both institutional and national platforms centered on patient-oriented research and aging-focused mentorship. As the co-PI of the Johns Hopkins AITC and the co-director of its Clinical Translation and Validation Core, Dr. Abadir actively contributes to the innovation of technologies tailored for older adults. Furthermore, he directs the molecular measurement core at the Older American Independence Center (OAIC) and is the associate director of the Translational Aging Research Training Program (T32). Mentoring Plan/Environment: Dr. Abadir's K24 proposal taps into the wealth of training assets at Johns Hopkins including resources from the JHU AITC, OAIC, the Institute for Clinical and Translational Research, and various T32 training grants. Special attention is given to mentoring investigators focused on populations at highest risk for aging-related conditions. Ideal mentees exhibit interest at the intersection of aging, technology, and Geroscience. The structured mentoring approach categorizes mentees based on experience, allocating specific effort percentages to ensure quality interactions. Regular individual meetings, hands-on research training, data interpretation, and presentation skills enhancement form core of mentoring strategy. Multi-tiered evaluations ensure consistent mentorship quality. This strategy aims to cultivate future leaders in translational aging research from both biological and engineering fields. Research Plan: The pioneering research funded by this K24 award aims to harness Artificial Intelligence (AI) tools and analytics for assessing physical and cognitive digital biosignals in frail older individuals including those with Alzheimer’s disease and associating these with molecular markers. The study will evaluate the accuracy and reliability of a new method that uses AI to analyze non-invasive multimodal biometric signals like speech, voice, eye movements, handwriting, and gait. This methodology will enhance our understanding of the varied characteristics of frailty in older individuals and pinpoint early cognitive shifts, including signs of Alzheimer’s. Building upon the existing efforts at Johns Hopkins AITC and OAIC, this research seeks to uncover new insights and broaden avenues for upcoming researchers.

PROJECT SUMMARY/ABSTRACT Celiac disease (CeD) is an autoimmune-like small intestinal enteropathy induced by dietary gluten in genetically susceptible individuals expressing HLA-DQ2 or HLA-DQ8 molecules. There is a spectrum in tissue damage with potential CeD patients developing inflammatory anti-gluten T cell immunity in the absence of tissue damage, while active CeD is associated with tissue damage ranging from partial to total villous atrophy. Gluten-free diet (GFD) is to this date the only treatment for CeD. However, 50% of CeD patients are unable to effectively adhere to a diet that sustainably excludes gluten, suffering therefore from inadvertent gluten exposure. Moreover, more than 30% of CeD patients have persistent high symptom burdens, resulting from continued mucosal damage despite adhering to a GFD. Persistent mucosal injury on a GFD is associated with several severe complications, including lymphoproliferative malignancy and bone diseases. Although much progress has been made in understanding the immunopathogenesis of CeD, biologic mechanisms underlying interindividual differences in clinical presentations and capacity to heal on a GFD are unknown. Qualitative and/or quantitative differences in the pool of pathogenic lymphocytes accumulating in the intestinal tissue could explain this heterogeneity. Despite mounting evidence from in vitro and murine models that a crosstalk between T cells, B cells, and plasma cells plays a critical role to increase the gluten-specific T cell response to a pathogenic threshold, whether T cell-B cell and T cell-plasma cell interactions contribute to enhancing the local T cell-mediated immune responses that have a direct impact on the formation of the gut celiac lesion has not been studied in humans. In addition, whether an altered mucosal suppressive CD4+ T cell response to gluten contributes to CeD pathogenesis is also unknown. The central hypothesis of this grant proposal is that pathogenic T cell clones responsible for the persistence of the gut celiac lesion and the inability of the tissue to heal are maintained through interactions with B cells and/or plasma cells or because of the inability of regulatory T cells to exert their suppressive function. Multiplex imaging and spatial transcriptomics coupled with B cell and T cell receptors repertoire analysis will be at the core of this proposal to determine lymphocyte clonal dynamics as well as cellular neighborhoods and spatial compartmentalization that support cellular interactions and their impact on tissue remodeling.

PROJECT SUMMARY/ABSTRACT The Pilot and Feasibility program at the University of Chicago (UChicago) Digestive Diseases Research Core Center (DDRCC) for Interdisciplinary Study of Inflammatory Intestinal Disorders (C-IID) is designed to provide a mechanism to promote new initiatives in digestive diseases (DD)-related research. The overall objectives of this program are to support investigations which would allow the development of preliminary data sufficient for grant funding by conventional mechanisms (pilot studies) or testing of an innovative hypothesis which might have important implications or yield significant results for digestive diseases related research (feasibility studies). Both of these broad aims have been pursued in the context of research focused on maintenance of homeostasis and an understanding of the pathobiology that accompanies intestinal inflammation. In the previous two renewal cycles (2010-2019), 45 P&F projects were awarded, and in the current period (2015-2019), 20 were awarded. The return on investment (ROI) over the past 10 years has exceeded 20:1. The P&F program remains an important vehicle for promoting multidisciplinary collaboration and a strong supportive scientific network for the young investigators funded by this program. Track I (new) investigators are given highest priority, followed by Tract II (non-DD) and Tract III (DD, new direction) investigators. We are requesting continuing support at the level of $110,000 per year. The funds will be combined with the $50,000 per year in supplemental funding committed by the GI section, bringing the total award amount to $160,000 or $30,000-37,500 for each award depending on the number of awards selected by the External Advisory Board. With the higher award size, the P&F awards are far more attractive to potential applicants and substantial enough to improve the odds that they will be successful in obtaining independent federal and extramural funding. A key feature of the program is the multi-disciplinary nature of the submitted proposals, coming from numerous departments, programs, and institutional affiliate within the University of Chicago network including the two of the University of Chicago affiliates, Argonne National Laboratory (ANL) and Marine Biological Laboratory (MBL). This large catchment of investigator talent reflects the overall scientific diversity of the research base of the C-IID. The P&F program has therefore made a difference for many investigators, fostered collaboration and interaction, pushed the envelope of scholarship and discovery, and is one of the jewels of our C-IID program.

PROJECT SUMMARY/ABSTRACT Celiac disease (CeD) is a complex intestinal inflammatory disorder that is triggered by dietary gluten and develops in genetically susceptible individuals expressing HLA-DQ2 or HLA-DQ8 molecules. 1% of the worldwide population is affected by this disease for which the only effective treatment is a lifelong and restrictive gluten-free diet (GFD). Yet, persistent symptoms and enteropathy remain commonplace even among CeD patients that adhere to a GFD. This stresses the need to develop non-dietary interventions for CeD. The development of new therapies has however proven challenging because of our incomplete understanding of the immune mechanisms underlying CeD pathogenesis and the lack of a suitable mouse model. CeD is characterized by the loss of oral tolerance to gluten manifested by HLA-DQ2 or HLA-DQ8- restricted anti-gluten inflammatory CD4 T cells in the small intestinal mucosa and by a massive expansion of cytotoxic intraepithelial CD8+ lymphocytes (IE-CTLs) that are involved in the killing of intestinal epithelial cells. These observations have led to the general idea that CeD is primarily a T cell-mediated immune disorder. We hypothesize, however, that B cells also play a critical role. This hypothesis stems from several observations. First, CeD is characterized by a considerable expansion of plasma cells in the mucosa of CeD patients as well as the development of anti-deamidated gluten peptides (DGP) antibodies and autoantibodies against the enzyme tissue transglutaminase 2 (TG2). Second, the main model to explain the production of anti-DGP and anti-TG2 antibodies is that gluten-specific CD4+ T cells provide help to B cells suggesting that B cells could act as antigen-presenting cells for T cells and promote the amplification of the anti-gluten CD4 T cell response. Finally, several case reports on patients having CeD associated with another autoimmune disease suggest that B cell depletion therapy can provide clinical benefit in CeD, and we have demonstrated that B cell depletion significantly reduces intestinal tissue damage in our mouse model of CeD. The objective of this application is to characterize in vivo the role of B cells in amplifying the anti-gluten T cell response and allow it to reach a sufficient magnitude to promote tissue destruction. This project is innovative as it employs unique mouse models of CeD allowing to manipulate B lymphocytes, gluten-specific T cells, the gluten antigen, and the CeD predisposing HLA molecule to 1) assess the contribution of B cells as antigen-presenting to the activation and amplification of the anti-gluten CD4+ T cell response, and 2) assess the role of B cells and antibodies in the activation of IE-CTLs and tissue destruction. The knowledge gained from this study will provide unprecedented insights into the mechanisms by which B cell-mediated immunity contribute to the pathogenesis of CeD and will assess for the first time the therapeutic potential of B cell depletion therapy in an experimental mouse model of CeD.

Chronic Obstructive Pulmonary Disease (COPD) is a leading cause of death. Increasing in prevalence, COPD is a major burden to patients and providers. Computed tomography (CT) can provide valuable information about the structural and functional abnormalities of the disease as demonstrated in numerous studies where quantitative CT is deployed to characterize and evaluate the treatment. For instance, the COPDGene study has recently shown the substantial role of quantitative CT in the redefinition of COPD diagnosis, and in evaluating the progression of emphysema over time. However, these biomarkers vary across different scanners, settings, and patient attributes. There is a crucial need to manage this variability by optimizing and harmonizing CT images for reliable biomarker quantifications across both current and emerging scanners. This goal is not possible through conventional methods of using physical phantoms or patient images. Physical phantoms are often oversimplified and not representative of the complex anatomy and physiology of COPD patients. Patient images are ground-truth-limited, i.e., the exact anatomy and physiology of the patient is not fully known. Further, patient-based comparisons require multiple acquisitions of the same subjects across different scanners and settings. This is not ethically possible since repeated imaging increases the absorbed radiation dose. These challenges can be overcome through the use of virtual imaging trials (VITs) where studies are performed in silico using computational models of patients and scanners. VITs can provide reliable and practical solution to the challenge of COPD imaging provided realistic models of patients and scanners. Such models are currently lacking in the context of COPD. We develop and then utilize realistic virtual imaging toolsets to systematically evaluate and optimize CT biomarkers in COPD patients across scanners, imaging parameters, and patient attributes. We develop the first library of realistic COPD patient models with diverse attributes and severities. Coupled with accurate models of different scanners, the phantoms will be used to generate sets of ground-truth-known virtual CT cases, to be disseminated to the research community and to be used to systematically evaluate the effects of current and emerging scanners, various patient attributes, and the effects of image processing algorithms (through a national challenge), on the accuracy and precision of COPD biomarkers. Further, we develop and optimize a truth-based artificial intelligence-based algorithm for COPD quantifications. We optimize the algorithm for accuracy and reproducibility, taking advantage of the ground-truth known simulated images . We then harmonize CT settings across different scanners to accurately and precisely assess COPD imaging biomarkers for both single time-point and longitudinal studies. The studies will be done for the top two image processing algorithms, identified in the challenge, as well as our developed algorithm. Through these efforts, the project will position CT as a more reliable method for improved characterization and monitoring of COPD.

The self-organization and spatial patterning of cells are critical for tissue morphogenesis and regeneration. While biochemical factors have traditionally been the focus of developmental and regenerative biology research, emerging studies suggest that tissue geometry plays a pivotal role in controlling cellular behavior and tissue function. Our research program investigates how macroscopic 3D tissue geometries—such as curvature, concavity, continuity, and aspect ratio (length-to-width ratio)—independently influence cell morphology, gene expression, and tissue homeostasis. The program will initially focus on human mesenchymal cells in skin and vascular tissues, addressing three fundamental questions: (1) How does tissue geometry influence steady-state cell morphology and transcriptional activity? (2) How do cells respond to dynamic changes in tissue geometry? (3) How are cellular signals propagated within tissue in response to changing geometric boundaries? To systematically decouple and investigate the effects of specific geometric cues in 3D, we will leverage our recently developed Edgeless Tissue Constructs (ETC)—a cutting-edge platform that allows precise control over tissue continuity, curvature, and concavity. Using this platform, we will examine cellular alignment, extracellular matrix organization, proliferation, migration, and shifts in transcriptional signatures in response to individual geometric cues under steady-state conditions. Additionally, we will integrate the ETC with robotic and automated systems to investigate how these dynamic 3D geometries—such as disrupted continuity (e.g., wounds) or increased curvature (e.g., aortic aneurysm)—affect cells independently of other microenvironmental factors like inflammation or parenchymal signals. Finally, we aim to uncover the biomolecular and biomechanical pathways by which cells sense new geometric boundary conditions—such as discontinuous boundaries—and propagate signals throughout the tissue. To achieve these goals, we will employ a comprehensive array of technologies, including single-cell and spatial transcriptomics (e.g., Xenium), computational modeling, mechanical testing (e.g., nanoindentation), confocal imaging (e.g., second-harmonic generation), and gene-editing tools. After establishing a strong foundation with human mesenchymal cells in the first five years, the program will expand to include other human cell types (e.g., epidermal, endothelial, and immune cells) and tissue types (e.g., uterus, adipose). Ultimately, this research may lead to the discovery of novel therapeutic strategies that target tissue geometry, as well as the development of more physiologically relevant organ models and grafts guided by geometric cues. Moreover, this interdisciplinary program will provide exceptional training opportunities for scientists, engineers, and clinicians at various educational levels by leveraging my established collaborations at Columbia University.

The self-organization and spatial patterning of cells are critical for tissue morphogenesis and regeneration. While biochemical factors have traditionally been the focus of developmental and regenerative biology research, emerging studies suggest that tissue geometry plays a pivotal role in controlling cellular behavior and tissue function. Our research program investigates how macroscopic 3D tissue geometries—such as curvature, concavity, continuity, and aspect ratio (length-to-width ratio)—independently influence cell morphology, gene expression, and tissue homeostasis. The program will initially focus on human mesenchymal cells in skin and vascular tissues, addressing three fundamental questions: (1) How does tissue geometry influence steady-state cell morphology and transcriptional activity? (2) How do cells respond to dynamic changes in tissue geometry? (3) How are cellular signals propagated within tissue in response to changing geometric boundaries? To systematically decouple and investigate the effects of specific geometric cues in 3D, we will leverage our recently developed Edgeless Tissue Constructs (ETC)—a cutting-edge platform that allows precise control over tissue continuity, curvature, and concavity. Using this platform, we will examine cellular alignment, extracellular matrix organization, proliferation, migration, and shifts in transcriptional signatures in response to individual geometric cues under steady-state conditions. Additionally, we will integrate the ETC with robotic and automated systems to investigate how these dynamic 3D geometries—such as disrupted continuity (e.g., wounds) or increased curvature (e.g., aortic aneurysm)—affect cells independently of other microenvironmental factors like inflammation or parenchymal signals. Finally, we aim to uncover the biomolecular and biomechanical pathways by which cells sense new geometric boundary conditions—such as discontinuous boundaries—and propagate signals throughout the tissue. To achieve these goals, we will employ a comprehensive array of technologies, including single-cell and spatial transcriptomics (e.g., Xenium), computational modeling, mechanical testing (e.g., nanoindentation), confocal imaging (e.g., second-harmonic generation), and gene-editing tools. After establishing a strong foundation with human mesenchymal cells in the first five years, the program will expand to include other human cell types (e.g., epidermal, endothelial, and immune cells) and tissue types (e.g., uterus, adipose). Ultimately, this research may lead to the discovery of novel therapeutic strategies that target tissue geometry, as well as the development of more physiologically relevant organ models and grafts guided by geometric cues. Moreover, this interdisciplinary program will provide exceptional training opportunities for scientists, engineers, and clinicians at various educational levels by leveraging my established collaborations at Columbia University.

PROJECT SUMMARY This application addresses targeted health issues through a Cooperative Research Agreement with the World Trade Center (WTC) Health Program (UO1). The presence of carcinogens and inducers of inflammation in WTC dust have raised the possibility that those exposed to WTC dust during the 9/11 attacks could have increased cancer incidence. To date, several cohort studies indicate that total cancer rates are 6-14% above background rates with significantly greater increases for thyroid and prostate cancer (Moir, 2016; Solan, 2013; Li, 2012; Jordan, 2011; Zeig-Owens, 2011). Increased exposure based on the time of arrival for workers, their proximity to early response to the WTC and duration of exposure have been associated with increased risk of tumor progression (de la Hoz, 2008; Lioy and Georgopoulos, 2006). Recent studies in small animal models indicate that WTC dust particles or metals are undetectable or present at exceedingly low levels in prostate tissues compared to lung following WTC dust exposure by inhalation or gavage (Wang, 2022; Gong, 2019), and long-term studies of rodents have not revealed evidence of a direct carcinogenic mechanism. Instead, there is evidence from both human RNA expression and DNA methylation analyses (Yu, 2022) and animal models (Wang, 2022; Gong, 2019) that inflammation induced by this dust may be implicated in promoting prostate cancer. In the mouse, WTC dust exposure promotes systemic as well as localized prostate inflammation, increased growth in PTEN deficient prostate epithelia and promotion of genetically initiated prostate tumors (Wang, 2022). Aim 1 will investigate whether WTC associated prostate cancer is more likely to recur independent of tumor grade at diagnosis and if there are molecular signatures of aggressiveness at diagnosis in those who recur. We will utilize increased biochemical (PSA) recurrence to define responders with adverse clinical response to WTC dust following surgery or radiation therapy. We will also apply recently identified signatures by RNA expression analysis and mass spec multiplex immunostaining of primary prostate cancer tissues to test whether these immune/inflammatory markers correlate with increased tumor recurrence in WTC responders. Aim 2 will focus on correlating our human findings with those in mouse genetically engineered mouse models bearing targeted lesions to prostate that model human PC. These experiments take advantage of our strong preliminary evidence that these models support a role of WTC dust exposure in PC progression. Experiments will include testing whether WTC dust exposure promotes recurrence of genetically initiated PCs following androgen deprivation therapy and/or experimental PC metastasis. We will also compare, and contrast WTC dust associated immune/inflammatory biomarkers in the mouse with those identified in human PCs. Our overarching goal is to translate understanding gained toward improved surveillance of WTC responders most at risk for prostate cancer.

This Career Development Award Level 2 (CDA2) represents the candidate’s effort to execute a pilot study of a novel neuromodulatory treatment for Veteran [social and occupational functioning], impulsivity and [suicidal ideation] following mild traumatic brain injury (mTBI). The candidate’s first aim will examine the tolerability, safety and feasibility of frontal pole intermittent theta burst stimulation (iTBS) for Veterans with mild traumatic brain injury (mTBI), negative urgency impulsivity, [and suicidal ideation]. This is important because impulsivity, which is exhibited in up to 40% of individuals with TBI, is an independent risk factor for dying by suicide and is associated with poorer functional outcomes following TBI. Presently, there are no widely accepted treatment strategies for impulsivity, [suicidal ideation and social and occupational functional deficits] following TBI. The candidate’s second aim is to determine the effects of frontal pole iTBS on social and community functioning, negative urgency impulsivity and [suicidal ideation]. This will allow the scientific community to better understand how frontal pole iTBS treatment impacts [social and community] functioning and mental health outcomes. Finally, the candidate aims to evaluate whether iTBS improves connectivity between the VMPFC and limbic system, using resting state functional connectivity MRI. Completing this aim will allow us to better understand how iTBS affects neural connectivity and whether these changes are related to functional and mental health outcomes. This research is critical as suicide is a major issue among Veterans within the VA system. Veterans are 22% more likely to die by suicide than civilians. Veterans with TBI are even more likely to die by suicide than those without TBI. It is important to create a treatment to prevent unnecessary Veteran deaths while helping Veterans feel empowered and functional within their communities. [Suicide and TBI are not unique to Veterans, however. Civilians with TBI are also more likely to die by suicide than those without. Thus, findings from this research could serve to improve neuropsychiatric care in the civilian sector, as well.] This project serves as a critical first step in allowing the candidate and her team to develop a biologically-informed treatment strategy for the many individuals who struggle with mTBI, [social and occupational functional deficits], impulsivity and [suicidal ideation]. Ultimately, the candidate’s long-term career goal is to become an independent clinical researcher in the VA system with expertise in TBI, suicidality, neurostimulation and neuroimaging. As such, she will complete a careful training plan under the mentorship of a strong, multidisciplinary training team involving opportunities for networking with experts in fields relating to the above research, hands-on training in necessary research skills and coursework complementary to the research project. The planned research is significant, innovative, and impactful, as it will be the first pilot study of a new iTBS treatment paradigm, designed to improve Veteran lives and functional outcomes after mTBI. It will also strengthen the candidate’s knowledge base and research skills, preparing her for an independent career in post-TBI rehabilitation research, with expertise in impulsivity, suicidality and neuromodulation.

This Career Development Award Level 2 (CDA2) represents the candidate’s effort to execute a pilot study of a novel neuromodulatory treatment for Veteran [social and occupational functioning], impulsivity and [suicidal ideation] following mild traumatic brain injury (mTBI). The candidate’s first aim will examine the tolerability, safety and feasibility of frontal pole intermittent theta burst stimulation (iTBS) for Veterans with mild traumatic brain injury (mTBI), negative urgency impulsivity, [and suicidal ideation]. This is important because impulsivity, which is exhibited in up to 40% of individuals with TBI, is an independent risk factor for dying by suicide and is associated with poorer functional outcomes following TBI. Presently, there are no widely accepted treatment strategies for impulsivity, [suicidal ideation and social and occupational functional deficits] following TBI. The candidate’s second aim is to determine the effects of frontal pole iTBS on social and community functioning, negative urgency impulsivity and [suicidal ideation]. This will allow the scientific community to better understand how frontal pole iTBS treatment impacts [social and community] functioning and mental health outcomes. Finally, the candidate aims to evaluate whether iTBS improves connectivity between the VMPFC and limbic system, using resting state functional connectivity MRI. Completing this aim will allow us to better understand how iTBS affects neural connectivity and whether these changes are related to functional and mental health outcomes. This research is critical as suicide is a major issue among Veterans within the VA system. Veterans are 22% more likely to die by suicide than civilians. Veterans with TBI are even more likely to die by suicide than those without TBI. It is important to create a treatment to prevent unnecessary Veteran deaths while helping Veterans feel empowered and functional within their communities. [Suicide and TBI are not unique to Veterans, however. Civilians with TBI are also more likely to die by suicide than those without. Thus, findings from this research could serve to improve neuropsychiatric care in the civilian sector, as well.] This project serves as a critical first step in allowing the candidate and her team to develop a biologically-informed treatment strategy for the many individuals who struggle with mTBI, [social and occupational functional deficits], impulsivity and [suicidal ideation]. Ultimately, the candidate’s long-term career goal is to become an independent clinical researcher in the VA system with expertise in TBI, suicidality, neurostimulation and neuroimaging. As such, she will complete a careful training plan under the mentorship of a strong, multidisciplinary training team involving opportunities for networking with experts in fields relating to the above research, hands-on training in necessary research skills and coursework complementary to the research project. The planned research is significant, innovative, and impactful, as it will be the first pilot study of a new iTBS treatment paradigm, designed to improve Veteran lives and functional outcomes after mTBI. It will also strengthen the candidate’s knowledge base and research skills, preparing her for an independent career in post-TBI rehabilitation research, with expertise in impulsivity, suicidality and neuromodulation.

Program Summary This study uses the natural laboratory of Nigeria to test the comparative effectiveness of two sets of implementation strategies to engage HIV patients in cervical cancer screening and treatments at scale. If successful, these implementation strategies can be adapted for engaging HIV patients in cervical cancer screening and treatment in low income and rural areas in the United States. Nigeria has the largest population of women and the 4th largest number of women living with HIV (WLHIV) in Africa. Although access to antiretroviral therapy (ART) among WLHIV in Nigeria has increased over the years, with over 98% of the 960,000 WLHIV on ART, AIDS-related mortality remains high. In 2020, 16,000 WLHIV died from AIDS related illnesses including cervical cancer. A pilot implementation program in Nigeria demonstrated that leveraging the U.S. President’s Emergency Plan for AIDS Relief (PEPFAR) supported HIV programs for the provision of evidence-based cervical cancer screen-and-treat interventions in WLWH is feasible. The pilot program demonstrated, however, that tailored implementation strategies are needed to address specific multilevel barriers along the cancer control continuum to address adoption, reach, and sustainment that are necessary for successful scale-up in low-resource settings. However, in regions of the United States with the burden of both HIV and cervical cancer, there is a paucity of evidence-based implementation strategies to inform effective integration of HIV and cervical cancer services delivery. Objectives of this proposal are to: 1) Refine strategies to integrate cervical cancer screening, treatment and management within existing comprehensive HIV treatment programs and determine implementation readiness; 2) Determine the comparative effectiveness of a Core set of implementation strategies versus Core+ enhanced implementation strategies; and 3) assess sustainment of the integration of cervical cancer screening, treatment, and management intervention into HIV programs. We have assembled a strong team from University of California San Diego; University of Nigeria, Nsukka, and Northeastern University with expertise in implementation science, HIV care and research, and cancer care and research. Our proposal is responsive to the NCI RFA and consistent with the World Health Organization global plan of elimination of cervical cancer by 2030. If effective, the proposed project will result in a set of feasible, culturally adaptable, and sustainable implementation strategies to integrate evidence-based cervical cancer screening and treatment into HIV programs in the United States in order to improve the health and life expectancy of WLHIV.

Program Summary This study uses the natural laboratory of Nigeria to test the comparative effectiveness of two sets of implementation strategies to engage HIV patients in cervical cancer screening and treatments at scale. If successful, these implementation strategies can be adapted for engaging HIV patients in cervical cancer screening and treatment in low income and rural areas in the United States. Nigeria has the largest population of women and the 4th largest number of women living with HIV (WLHIV) in Africa. Although access to antiretroviral therapy (ART) among WLHIV in Nigeria has increased over the years, with over 98% of the 960,000 WLHIV on ART, AIDS-related mortality remains high. In 2020, 16,000 WLHIV died from AIDS related illnesses including cervical cancer. A pilot implementation program in Nigeria demonstrated that leveraging the U.S. President’s Emergency Plan for AIDS Relief (PEPFAR) supported HIV programs for the provision of evidence-based cervical cancer screen-and-treat interventions in WLWH is feasible. The pilot program demonstrated, however, that tailored implementation strategies are needed to address specific multilevel barriers along the cancer control continuum to address adoption, reach, and sustainment that are necessary for successful scale-up in low-resource settings. However, in regions of the United States with the burden of both HIV and cervical cancer, there is a paucity of evidence-based implementation strategies to inform effective integration of HIV and cervical cancer services delivery. Objectives of this proposal are to: 1) Refine strategies to integrate cervical cancer screening, treatment and management within existing comprehensive HIV treatment programs and determine implementation readiness; 2) Determine the comparative effectiveness of a Core set of implementation strategies versus Core+ enhanced implementation strategies; and 3) assess sustainment of the integration of cervical cancer screening, treatment, and management intervention into HIV programs. We have assembled a strong team from University of California San Diego; University of Nigeria, Nsukka, and Northeastern University with expertise in implementation science, HIV care and research, and cancer care and research. Our proposal is responsive to the NCI RFA and consistent with the World Health Organization global plan of elimination of cervical cancer by 2030. If effective, the proposed project will result in a set of feasible, culturally adaptable, and sustainable implementation strategies to integrate evidence-based cervical cancer screening and treatment into HIV programs in the United States in order to improve the health and life expectancy of WLHIV.

Program Summary Nigeria has one of the largest HIV epidemics in the world with 1.8 million people living with HIV infection. With an estimated female population of 102 million and HIV prevalence of 1.6% among adult females, Nigeria has the largest population of women and the 4th largest number of women living with HIV (WLHIV) in Africa. Although access to antiretroviral therapy (ART) among WLHIV in Nigeria has increased over the years, with over 98% of the 960,000 WLHIV on ART, AIDS-related mortality remains high. In 2020, 16,000 WLHIV died from AIDS- related illnesses including cervical cancer. A pilot implementation program in Nigeria demonstrated that leveraging the U.S. President’s Emergency Plan for AIDS Relief (PEPFAR) supported HIV programs for the provision of evidence-based cervical cancer screen-and-treat interventions in WLWH is feasible. The pilot program demonstrated, however, that tailored implementation strategies will be needed to address specific multilevel barriers along the cancer control continuum in order to address adoption, reach, and sustainability that are necessary for successful scale-up. However, in many African countries with a high burden of both HIV and cervical cancer, there is a paucity of evidence-based implementation strategies to inform effective integration of HIV and cervical cancer services delivery. Objectives of this proposal are to: 1) Refine strategies to integrate cervical cancer screening, treatment and management within existing comprehensive HIV treatment programs and determine implementation readiness; 2) Determine the comparative effectiveness of a Core set of implementation strategies versus Core+ enhanced implementation strategies; and 3) assess sustainment of the integration of cervical cancer screening, treatment, and management intervention into HIV programs. We have assembled a strong team from University of California San Diego; University of Nigeria, Nsukka and John Hopkins University with expertise in implementation science, HIV care and research, and cancer care and research. Our proposal is responsive to the NCI RFA and consistent with the World Health Organization global plan of elimination of cervical cancer by 2030. If effective, the proposed project will result in a set of feasible, culturally adaptable, and sustainable implementation strategies to integrate evidence-based cervical cancer screening and treatment into HIV programs in order to improve the health and life expectancy of WLHIV.

PROJECT SUMMARY (Methods Core) Team approaches to meet organizational goals are utilized broadly and globally in industry, the military, space exploration, and health settings. The TEAMS Methods Core is integral to adapting and advancing team-based models, approaches, strategies, methods, and evaluation for children’s mental health and for the larger center activities. Of note, the Methods Core will lead innovation by leveraging natural language processing (NLP) technology to develop tools to rapidly assess team functioning in real-world settings. Within community service systems caring for children with mental health needs, the assessment of team effectiveness—although crucial for improving team functioning—can be a laborious and cumbersome task. Innovations in technology related to the automatic capturing of communication between team members can lessen the burden in assessing and improving team effectiveness. The Methods Core will lead innovation by leveraging natural language processing technology to develop tools to rapidly assess team functioning in real-world settings. The Methods Core investigators have already been collaborating closely with the Administrative Core, the R01, and the R34 teams, and Community Service System Workgroup stakeholders in selecting, adapting and shaping strategies and approaches to implementation and improvement for children’s mental health across service system settings. The Methods Core is a team effort led by Drs. Aarons (implementation science, mental health), Burke (team effectiveness, team strategies), and Sklar (mental health services, evaluation) (see biosketches). The Methods Core is organized to address four aims: Aim 1) Integrate models and strategies from team effectiveness research and implementation science to improve children’s mental health services, Aim 2) Apply multilevel and advanced methods and designs for team-based implementation research, and Aim 3) Lead the design and implementation of a multi-level, longitudinal, iterative evaluation to refine the Center’s research activities, maximize the Center’s impact and inform plans for future collaborative research.

Program Summary Nigeria has one of the largest HIV epidemics in the world with 1.8 million people living with HIV infection. With an estimated female population of 102 million and HIV prevalence of 1.6% among adult females, Nigeria has the largest population of women and the 4th largest number of women living with HIV (WLHIV) in Africa. Although access to antiretroviral therapy (ART) among WLHIV in Nigeria has increased over the years, with over 98% of the 960,000 WLHIV on ART, AIDS-related mortality remains high. In 2020, 16,000 WLHIV died from AIDS- related illnesses including cervical cancer. A pilot implementation program in Nigeria demonstrated that leveraging the U.S. President’s Emergency Plan for AIDS Relief (PEPFAR) supported HIV programs for the provision of evidence-based cervical cancer screen-and-treat interventions in WLWH is feasible. The pilot program demonstrated, however, that tailored implementation strategies will be needed to address specific multilevel barriers along the cancer control continuum in order to address adoption, reach, and sustainability that are necessary for successful scale-up. However, in many African countries with a high burden of both HIV and cervical cancer, there is a paucity of evidence-based implementation strategies to inform effective integration of HIV and cervical cancer services delivery. Objectives of this proposal are to: 1) Refine strategies to integrate cervical cancer screening, treatment and management within existing comprehensive HIV treatment programs and determine implementation readiness; 2) Determine the comparative effectiveness of a Core set of implementation strategies versus Core+ enhanced implementation strategies; and 3) assess sustainment of the integration of cervical cancer screening, treatment, and management intervention into HIV programs. We have assembled a strong team from University of California San Diego; University of Nigeria, Nsukka and John Hopkins University with expertise in implementation science, HIV care and research, and cancer care and research. Our proposal is responsive to the NCI RFA and consistent with the World Health Organization global plan of elimination of cervical cancer by 2030. If effective, the proposed project will result in a set of feasible, culturally adaptable, and sustainable implementation strategies to integrate evidence-based cervical cancer screening and treatment into HIV programs in order to improve the health and life expectancy of WLHIV.

PROJECT SUMMARY (Methods Core) Team approaches to meet organizational goals are utilized broadly and globally in industry, the military, space exploration, and health settings. The TEAMS Methods Core is integral to adapting and advancing team-based models, approaches, strategies, methods, and evaluation for children’s mental health and for the larger center activities. Of note, the Methods Core will lead innovation by leveraging natural language processing (NLP) technology to develop tools to rapidly assess team functioning in real-world settings. Within community service systems caring for children with mental health needs, the assessment of team effectiveness—although crucial for improving team functioning—can be a laborious and cumbersome task. Innovations in technology related to the automatic capturing of communication between team members can lessen the burden in assessing and improving team effectiveness. The Methods Core will lead innovation by leveraging natural language processing technology to develop tools to rapidly assess team functioning in real-world settings. The Methods Core investigators have already been collaborating closely with the Administrative Core, the R01, and the R34 teams, and Community Service System Workgroup stakeholders in selecting, adapting and shaping strategies and approaches to implementation and improvement for children’s mental health across service system settings. The Methods Core is a team effort led by Drs. Aarons (implementation science, mental health), Burke (team effectiveness, team strategies), and Sklar (mental health services, evaluation) (see biosketches). The Methods Core is organized to address four aims: Aim 1) Integrate models and strategies from team effectiveness research and implementation science to improve children’s mental health services, Aim 2) Apply multilevel and advanced methods and designs for team-based implementation research, and Aim 3) Lead the design and implementation of a multi-level, longitudinal, iterative evaluation to refine the Center’s research activities, maximize the Center’s impact and inform plans for future collaborative research.

PROJECT SUMMARY The long-term goal of this K23 Career Development Award is to prepare the PI (Rachel Aaron, PhD) for an independent research career that aims to promote adaptive recovery for individuals who survive orthopedic trauma. Orthopedic trauma, resulting in severe injuries such as multiple fractures or amputation, occurs in around 3 million people annually in the United States; about half of survivors experience persistent pain and psychological distress in the year following injury, and most report substantial disability 7 years post trauma. There is an urgent need to identify factors that underlie pain and psychological distress following orthopedic trauma and to develop targeted psychological interventions to treat these potentially disabling symptoms. The broader literature suggests that difficulties with emotion regulation (i.e., identifying one's emotions and engaging in strategies to up- or down-regulate them) and central sensitization (i.e., alterations in the endogenous modulation of pain) lead to poor pain-related outcomes. Using a theory-driven assessment of emotion regulation, and gold-standard, laboratory-based assessment of central sensitization (quantitative sensory testing [QST]), Study 1 will examine emotion regulation difficulties and central sensitization at 6 weeks post trauma as predictors of persistent pain, distress, and opioid use 6 months post trauma. Study 2 will test the feasibility of assessing and delivering Emotional Awareness and Expression Therapy (EAET) to people who endorse persistent pain (i.e., clinically significant pain, present most days for the past 3 months) at 6 months post trauma. EAET is a novel chronic pain psychology intervention that treats pain and distress by targeting trauma-related emotion regulation process. Trauma exposure is ubiquitous among orthopedic trauma survivors; thus, EAET may be ideally suited for those with persistent pain post trauma. However, the feasibility of delivering EAET is unclear due to established barriers engaging this population in mental health treatment. Proposed research and career development activities will take place at Johns Hopkins University (JHU), a renowned research environment. JHU is dedicated to the success of its junior faculty and provides unique access to orthopedic trauma patients. The PI has formed a strong team of content experts in central sensitization (including QST), psychological intervention (including EAET), and orthopedic trauma. Immersed in JHU's rich training environment, the PI will achieve the following career goals through dedicated mentorship, didactic experiences, and professional development opportunities: (1) develop expertise in the assessment of central sensitization, including QST; (2) gain foundational skills for intervention development, implementation, and analysis; and (3) learn to lead impactful orthopedic trauma clinical research. The completion of this K23 proposal will provide the necessary training and preliminary data for the PI to obtain independent grant funding and systematically pursue a line of research to improve outcomes for survivors of orthopedic trauma.

PROJECT SUMMARY The long-term goal of this K23 Career Development Award is to prepare the PI (Rachel Aaron, PhD) for an independent research career that aims to promote adaptive recovery for individuals who survive orthopedic trauma. Orthopedic trauma, resulting in severe injuries such as multiple fractures or amputation, occurs in around 3 million people annually in the United States; about half of survivors experience persistent pain and psychological distress in the year following injury, and most report substantial disability 7 years post trauma. There is an urgent need to identify factors that underlie pain and psychological distress following orthopedic trauma and to develop targeted psychological interventions to treat these potentially disabling symptoms. The broader literature suggests that difficulties with emotion regulation (i.e., identifying one's emotions and engaging in strategies to up- or down-regulate them) and central sensitization (i.e., alterations in the endogenous modulation of pain) lead to poor pain-related outcomes. Using a theory-driven assessment of emotion regulation, and gold-standard, laboratory-based assessment of central sensitization (quantitative sensory testing [QST]), Study 1 will examine emotion regulation difficulties and central sensitization at 6 weeks post trauma as predictors of persistent pain, distress, and opioid use 6 months post trauma. Study 2 will test the feasibility of assessing and delivering Emotional Awareness and Expression Therapy (EAET) to people who endorse persistent pain (i.e., clinically significant pain, present most days for the past 3 months) at 6 months post trauma. EAET is a novel chronic pain psychology intervention that treats pain and distress by targeting trauma-related emotion regulation process. Trauma exposure is ubiquitous among orthopedic trauma survivors; thus, EAET may be ideally suited for those with persistent pain post trauma. However, the feasibility of delivering EAET is unclear due to established barriers engaging this population in mental health treatment. Proposed research and career development activities will take place at Johns Hopkins University (JHU), a renowned research environment. JHU is dedicated to the success of its junior faculty and provides unique access to orthopedic trauma patients. The PI has formed a strong team of content experts in central sensitization (including QST), psychological intervention (including EAET), and orthopedic trauma. Immersed in JHU's rich training environment, the PI will achieve the following career goals through dedicated mentorship, didactic experiences, and professional development opportunities: (1) develop expertise in the assessment of central sensitization, including QST; (2) gain foundational skills for intervention development, implementation, and analysis; and (3) learn to lead impactful orthopedic trauma clinical research. The completion of this K23 proposal will provide the necessary training and preliminary data for the PI to obtain independent grant funding and systematically pursue a line of research to improve outcomes for survivors of orthopedic trauma.

The purpose of the T35 Training Grant titled, Enhancement of Optometric Education with Basic, Applied and Clinical Research, is to afford qualified Optometry Interns the opportunity to pair with experienced research faculty to conduct basic, applied and clinical research at the University of the Incarnate Word Rosenberg School of Optometry (UIWRSO). Objectives include (a) Faculty-Intern research committed to early detection, diagnosis, and treatment of ocular disease as well as basic and applied research which enhance our understanding of visual function leading to improvement of vision in normal and abnormal populations (b) inspire Interns to pursue postdoctoral research and academic activities and positions paving a course toward academic and research careers (c) foster collaboration with other UIW programs (Schools of Medicine, Pharmacy, Rehabilitation Sciences, Nursing, Math, Science and Engineering and Biomedical Sciences) as well as institutions and industry sharing common goals (d) Increase research involvement of junior and clinical Faculty to enhance personal and professional development. The rationale for our proposal is predicate on our success with our Summer Fellowship Training Program (SFTP), active since 2011, allowing Interns to participate in faculty supervised research during the summer between first and second professional years. SFTP has been highly successful yielding 11 peer-reviewed papers and 50 peer-reviewed poster and paper presentations (7 by Interns) at major meetings, exceeding some state supported major schools and colleges of optometry funded with T35 grants. While the SFTP is largely internally funded, supplemented by industry and a grant to enhance optometric education, it closely mirrors current T35 summer programs; we will replicate and enhance our success pending award of a T35 Grant. We propose that a T35 grant support seven Interns per year for 8-week summer periods for a duration of five years. We anticipate continuing the success documented in our Program Plan, with numerous Interns presenting at major meetings and contributing to peer-reviewed publications, while also elevating the involvement of our junior Faculty in research activities and encouraging Interns to pursue academic and research careers.

Administrative Supplement Application Abstract/Summary The Whitman-Walker Institute Biomedical Research Hub that this supplement will support, represents a transformative investment in community-based biomedical research infrastructure that integrates advanced clinical diagnostics, bio specimen science, investigational pharmacy, implementation science, and clinical research within a trusted healthcare organization serving Washington, DC. Designed as an extension of the parent C06 award, the Biomedical Research Hub will create complementary research facilities that expand institutional capacity to conduct innovative HIV research while improving access to scientific advances for communities disproportionately affected by HIV. The Biomedical Research Hub will encompass the complete continuum of HIV clinical research. Beginning with patient evaluation and clinical care, the facility will support specimen collection, laboratory processing, tissue culture, molecular diagnostics, biospecimen preservation, investigational pharmacy operations, implementation science, and clinical trials. Together these complementary capabilities create a research environment capable of supporting investigator-initiated studies, federally funded clinical trials, translational science, community-initiated research, implementation research, and future biomedical innovation. The Biomedical Research Hub consists of two distinct but complementary research infrastructure projects and facilities that will increase Whitman-Walker Institute's research capacity - the Clinical Diagnostics and Biospecimen Processing Core, and the Clinical Pharmacy and Therapeutics Core. Although each facility serves a distinct scientific function, together they establish an integrated Biomedical Research Hub capable of supporting increasingly sophisticated HIV research and related comorbidities while remaining fully aligned with the objectives of the parent C06 award. We are requesting grant suppo1t to complete the construction of the Clinical Pharmacy and Therapeutics Core centered on USP 797-compliant sterile compounding, investigational product management, medication accountability, pharmacy support for clinical trials, and implementation of innovative HIV therapeutic strategies. Goal 1: Build-out a portion of a 6,000 sq. ft, section of the 3rd floor at 1201 Sycamore Drive SE, Washington, DC, 20032 to include: • 1a: A research pharmacy of approximately 550 sq.ft. compliant with USP-797 regulations to build capacity for non-hazardous sterile compounding drug trials sponsored by federal and industry partners.

Administrative Supplement Application Abstract/Summary The Whitman-Walker Institute Biomedical Research Hub that this supplement will support, represents a transfonnative investment in community-based biomedical research infrastructure that integrates advanced clinical diagnostics, bio specimen science, investigational pharmacy, implementation science, and clinical research within a trusted healthcare organization serving Washington, DC. Designed as an extension of the parent C06 award, the Biomedical Research Hub will create complementary research faci lities that expand institutional capacity to conduct innovative HIV research while improving access to scientific advances for communities disproportionately affected by HIV. The Biomedical Research Hub will encompass the complete continuum of HIV clinical research. Beginning with patient evaluation and clinical care, the facility will support specimen collection, laboratory processing, tissue culture, molecular diagnostics, biospecimen preservation, investigational pharmacy operations, implementation science, and clinical trials. Together these complementary capabilities create a research environment capable of supporting investigator-initiated studies, federally funded clinical trials, translational science, community-initiated research, implementation research, and future biomedical innovation. The Biomedical Research Hub consists of two distinct but complementary research infrastructure projects and faci lities that will increase Whitman-Walker Institute's research capacity - the Clinical Diagnostics and Biospecimen Processing Core, and the Clinical Pharmacy and Therapeutics Core. Although each facility serves a distinct scientific function, together they establish an integrated Biomedical Research Hub capable of supporting increasingly sophisticated HIV research and related comorbidities while remaining fully aligned with the objectives of the parent C06 award. We are requesting grant suppo1t to complete the construction of the Clinical Diagnostics and Biospecimen Processing Core dedicated to clinical specimen acquisition, laboratory processing, tissue culture, cryogenic storage, molecular diagnostics, and biospecimen banking. Goal 1: Build-out a po1tion of a 6,000 sq. ft, section of the 3rd floor at 1201 Sycamore Drive SE, Washington, DC, 20032 to include: • la: A research laboratory and secure storage of approximately 1,300 sq.ft. compliant with Biosafety Level 2 (BSL-2), to build capacity for translational HIV co-infections and comorbidities research. • 1 b: Approximately 1,400 sq.ft. of consult, exam, procedure, and infusion rooms for participant-facing research and approximately 2,750 sq.ft. of faculty and staff offices to expand our current capacity.