Johns Hopkins University

NIH Research Projects · FY 2024 · 2024-07

Project Summary (Abstract) This two-year grant proposal responds to PAR-23-179, and its long-term goal is to advance the understanding of biomarker in relation to preclinical Alzheimer’s Disease (AD). This proposal seeks to leverage interpretable and flexible tree-based methods, clinically and biologically informed tree decision rules, longitudinal biomarkers, and risk and protective factors, to develop novel statistical methods and construct tree-based algorithms. The novel methods and tree-based algorithms will help identify time-dynamic and personalized biomarker subgroups at high risk for cognitive decline due to AD and predict progression risks. AD is a devastating disease affecting over 6 million people in the U.S. and has burdened the U.S. healthcare system and caregivers with increases. Importantly, evidence suggests that the pathophysiological process begins many years, if not decades, before the diagnosis of AD dementia, and recent findings demonstrate that biomarker deterioration starts many years before cognitive decline due to AD. Identifying high-risk subgroups based on biomarker information will expand the window of opportunity during which therapeutic intervention may have the greatest potential for success. Tree-based methods appear well-suited for producing clinically applicable decision rules that leverage complicated interactions between different biomarkers, and between biomarkers and risk factors. But existing tree-based methods are limited in clinical relevance, biological interpretability, and statistical inference, and novel methods are needed. Meanwhile, the consortium of multiple longitudinal follow-up studies presents new opportunities and challenges. The Preclinical AD Consortium (PAC) data comprises five studies that have been collecting longitudinal clinical, cognitive, biomarker, and genetic data from individuals who were cognitively normal when first enrolled and followed for many years. The large sample size and the breadth of the merged and harmonized data create opportunities for more precise and personalized classification and risk prediction, based on longitudinal biomarkers and risk and protective factors. These opportunities also create challenges for method development to be time-dynamic and personalized. Projects supported by this proposal will seek to develop novel tree-based statistical methods for subgroup identification and risk prediction for cognitive decline due to AD, and construct classification and prediction algorithms using the PAC data. Our research team will pursue these goals through two Specific Aims: (1) establish a time-dynamic and personalized statistical classification and prediction framework using tree-based methods; and (2) leverage the PAC data to construct classification and prediction algorithms for onset of AD- related clinical symptoms. Successful completion of these Aims will produce novel tree-based methodologies and generate tree-based classification and prediction algorithms that predict onset of AD-related symptoms.

NSF Awards · FY 2024 · 2024-07

Topology and geometry in low dimensions differ significantly from those in the stable, high-dimensional, range. One feature of low dimensions is the existence of deep structures known as TQFTs (Topological Quantum Field Theories), many originating in quantum physics and having applications to condensed matter, statistical mechanics and quantum field theory. The Prinicipal Investigator will be studying such topological theories of more general type, where the theory is known on topological objects without boundary (closed objects) and extended canonically to objects with boundary. These constructions proved fruitful for explicit combinatorial construction of link homology theories, where topological objects are foam-like two-dimensional structures embedded in 3-space. The author has recently shown that a semi-linear version of this construction in dimension one extends so-called finite state automata and regular languages, which is a classical subject in computer science. This opens possibility of many generalizations, including exploring connections between more general languages and topological theories and possible relations between two-dimensional theories and cellular automata. Further studies of topological theories and related topics of foams and link homology should lead to fruitful discoveries in low-dimensional topology and related fields. More specifically, the project has three major goals. The first major goal is to further develop the theory of foams, their evaluations and applications in link homology and categorification. Foams are two-dimensional CW-complexes with generic singularities. They have proved instrumental in combinatorial approaches to GL(N) link homology theories and boast tantalizing connections to instanton Floer homology for orbifolds. The PI will further develop topological theories related to foams, with an eye towards technically difficult problems, such as computation of Kronheimer-Mrowka homology of embedded trivalent graphs and finding combinatorial counterpart of that homology. The second goal is to find approaches to several link homology theories, including Cautis, Webster and Qi-Sussan homologies, to establish their functoriality and extend to tangles and tangle cobordisms. A number of important link homology theories, including triply-graded HOMFLYPT homology, Webster, Cautis, and Qi-Sussan homology, are missing a functorial extension to tangle cobordisms and, in most cases, a related extensions to tangles. The PI will develop new approaches to these homology theories to redefine them, repair functoriality where necessary, and extend them to link cobordisms. The third goal is to understand universal theories in low dimensions. The PI will continue studying universal construction of topological theories, motivated by recent successes such as the interpretation of finite state automata and regular languages via one-dimensional topological theories with defects and taking values in the Boolean semiring B, where a regular language and a circular regular language give rise to a rigid symmetric monoidal B-linear category. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-07

ABSTRACT Indigenous peoples in the United States and worldwide face the starkest inequities in premature mortality of any racial and ethnic group. In the United States, American Indian/ Alaska Native (AI/AN) Peoples have 2X higher rates of suicide, 4X higher rates of alcohol induced deaths, and 1.6X higher drug overdose deaths compared to the US general population. To effectively drive change in the staggering inequities, we need a scientific workforce that better represents Indigenous peoples. To date, Indigenous and other diverse scholars are vastly underrepresented in NIH funding, including fellowship and training grants. The parallel inequities in public health education, training, and preventable cause mortality stem from historical and contemporary social and structural determinants. The goal of the Elevating Indigenous Wellbeing through Assets-Based Prevention Science (ELEVATE) Training Program is to train Indigenous and other diverse predoctoral scholars to become leaders in health equity research and strengths-based multi-level prevention science, addressing worsening inequities in mental- and behavioral-health-related premature mortality among Indigenous Peoples. To accomplish this goal, we will provide rigorous training and high-quality mentorship in (1) Health disparities/health equity research, (2) Developing and implementing multi-level prevention interventions, and (3) Methods to design and analyze studies that evaluate multi-level prevention interventions. We will also provide cross-cutting training in (4) Indigenous methodologies as a way to root this training and decolonizing approaches. Trainees well benefit from the program’s interdisciplinary and team-based mentorship and training approach. The ELEVATE program will he led by a team of Indigenous and allied leaders and leverage the vast resources of the Johns Hopkins Blumberg School of Public Health, including the Center for Indigenous Health’s (CIH) 40+ year trust-relationships with Tribal Nations and growing number of Indigenous and allied faculty. Specifically, the ELEVATE program will be housed in the Social and Behavioral Interventions program, but the 3 trainees per year may also come from the Department of Mental Health or full-time students from the school wide Doctorate of Public Health Program. Trainees well undertake a rigorous program of coursework in health equity, multilevel prevention, and associated and Indigenous methodologies. A year-long seminar to discuss research in progress, ongoing mentored research projects, and integrative activities will complement coursework. ELEVATE will synergize with CIH’s NIDA-funded P50 National Center for Excellence, Community-Driven Indigenous Research, Cultural Strengths & Leadership to Advance Equity in Drug Use Outcomes (CIRCLE). This proposed program aligns directly with the goal of the ADVANCE Predoctoral T32, the cross-cutting themes for the Office of Disease Prevention’s strategic plan, and would address key cross-cutting strategic priorities for NIDA, NIMH, and NIAAA. There are currently no predoctoral training programs for Indigenous health. The ELEVATE program fills this gap and will help build the next generation of Indigenous and other diverse prevention scientists.

NSF Awards · FY 2024 · 2024-07

NON-TECHNICAL SUMMARY: Many elements play essential roles in the advancement of technology. Critical elements, including rare earth metals, are vital for electronics, renewable energy, and advanced materials. However, the stable supply of these elements remains a challenge due to the limitations of the current extraction and separation methods that are high-cost, polluting, and/or geographically constrained. Nature has evolved sophisticated metal-binding peptides (MBPs) that selectively recognize and bind certain ions and molecules, which show great promise as a new modality for critical element separation. In practice, these MBPs need to be immobilized in a porous scaffold to facilitate the adsorption of targeted elements and promote the recycling of MBPs. The goal of this project is to study and develop a new class of immobilization substrates, namely mesoporous protein crystals, for critical element separation. Protein crystals show several appealing properties for MBP immobilization, including appropriate pore size and distribution, structural stability, and low toxicity, but they are limited by a low propensity of crystal nucleation. In this project, computationally designed protein crystals will be used to study the nucleation and growth behaviors of protein crystals. The structure-property relationship of protein crystals will be systemically characterized. The team will further evaluate immobilization strategies to maximize the density of MBPs in crystals while maintaining a sufficient porosity for ion diffusion. The capability of immobilized MBPs in sequestering critical elements will be evaluated and compared to the free MBPs. The success of the project will create a new MBP immobilization platform for efficient and selective critical element separation. The platform can be generalized to immobilize other materials for broader applications. Additionally, the project will provide valuable training and education opportunities to graduate, undergraduate, and high-school students by developing educational modules on advanced biomaterials for energy and sustainability. TECHNICAL SUMMARY: This project aims to obtain a comprehensive understanding of protein crystals as immobilization substrates for critical element separation. The team will combine protein engineering and rational design to tune the assembly of protein crystals and unravel factors that impact the crystal nucleation and growth. Batch crystallization will be exploited for the scalable synthesis of protein crystals and their physicochemical properties in varied structures and binding affinity will be systemically investigated. The team will explore immobilization strategies that can maximize the gravimetric ratio of guest MBP proteins in the crystalline scaffold while maintaining stable immobilization without gradual loss of MBPs over time. The immobilization platform will be examined by immobilizing Lanmodulin (LanM), a protein derived from nature for selective sequestration of rare earth elements (REEs). The capability of LanM-crystal complexes in binding REEs and their selectivity over other bivalent and trivalent cations will be evaluated. The team will further test the performance of immobilized LanM in practical REE extraction scenarios, such as extraction from simulated low-grade leachate using fixed-bed columns packed with LanM-crystal complexes. The new immobilization platform will address the scalability, cost, and stability issues of protein-based adsorbents that traditionally hinder their deployment. The success of this project will also offer a secure and resilient critical elements supply chain essential to economic prosperity and national defense. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-07

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. The otolaryngology specialty requires a wide variety of expertise and scientific strategies for understanding and treating communication disorders. Our program’s goal is to train and develop physicians who will advance the field of otolaryngology and serve as its future innovators and leaders. We aim to provide a solid foundation for medical student and resident participants’ futures in otolaryngology research by 1) providing tailored mentored research experiences for otolaryngology residents and medical students interested in matching with otolaryngology residency programs; 2) leveraging our cross-disciplinary institutional resources to provide professional development training and research skill-building to prepare participants for clinician-scientist careers; and 3) enhancing recruitment of individuals with primarily clinical backgrounds to careers in otolaryngology research. We will provide the research training, communication, and professional skills that will enable our graduates to become creative contributors to the future of otolaryngology and the treatment of associated communication disorders. Two residents per year will enter into 18 continuous months of research training in their third year of residency. Additionally, 2 medical students per year will be recruited for 9 months of research training. These students will be paired with near-peer resident mentors. These trainees can choose from a wide and deep selection of research themes both within the department and in associated laboratories at Hopkins and elsewhere. Mentoring teams will include preceptors and co-preceptors with a history of training clinician- scientists and early healthcare professionals, academic achievement, and competing for research funding. Research topics include, but are not limited to: basic mechanisms of, and therapeutic innovation for dizziness and balance; studies of the auditory nervous system; pathogenesis of sinusitis, laryngotracheal stenosis and respiratory papillomatosis; outcomes, quality and safety in otolaryngology; molecular biology and epidemiology of head and neck cancers; surgical robotics; and public health studies in otolaryngology-head and neck surgery. Program participants will benefit from access to state-of-the-art research facilities, a highly collaborative research environment representing a variety of disciplines, and substantial institutional resources. Participants will also complete a curriculum focused on safety and compliance, rigor & reproducibility, research skill-building, grantsmanship, laboratory management, and navigating careers as clinician- scientists. The program will build upon 33 years of successful research training in our T32 program, which has produced many clinician-investigators and leaders in otolaryngology-head and neck surgery.

NIH Research Projects · FY 2024 · 2024-07

Project Summary This major instrumentation proposal requests funds for the purchase of a JEOL 500 MHz NMR spectrometer to support a large user base of NIH-funded researchers at the Johns Hopkins School of Medicine (JHSOM). Re- searchers supported by the JHSOM NMR facility come from diverse clinical and basic science departments and are funded by numerous NIH institutes (NIGMS, NCI, NIAID, NIDDK, NIDA, NINDS, NIA, NIBIB, NEI, and NIMH). This new state-of-the-art spectrometer package consists of a 11.7T, shielded, 54 mm bore magnet, a spectrom- eter console with an RF generation system, preamplifiers, power amplifiers, digital receivers, and a field gradient system. The requested 5 mm FG/RO HFX digital autotune probe and 24 slots autosampler are essential features to support characterization of small molecules studied by our user base and to maximize throughput and ease- of-use. The new Dell Precision Workstation and the pulse programmer controlled by the Delta FT-NMR software provide a friendly user interface that will facilitate the achievement of research goals. Since the previous sub- mission, due to frequent and expensive repairs and crippling service disruptions, our previous 500 MHz Bruker system was replaced with the JEOL 500 MHz spectrometer described above. We are currently using the JEOL system under a negotiated evaluation agreement with an option to pay and own, extend the lease, or return without full payment by May 10, 2024. For 23 years this NMR core resource, which is managed and supported by the Department of Pharmacology and Molecular Sciences (DPMS), has been key in supporting biomedical chemistry research at the school of medicine. The health-related goals of the supported projects range from development of new imaging agents, new drug discovery for cancer and neurological diseases, and basic inves- tigations into cellular metabolomics and drug metabolism. The facility provides an essential resource for syn- thetic chemistry, drug development and basic science research efforts at the medical school campus. It provides the main tool for our chemists to assess the outcome of chemical transformations (1D 1H, 31P, 13C and 19F NMR spectra). Our research spans all classes of organic molecules of biomedical interest, including bioactive probes, drugs, and polymer conjugated molecules. In addition, the capabilities of this moderately high-field instrument allow for more sophisticated homonuclear and heteronuclear experiments to elucidate the structure of challeng- ing small molecules, nucleic acids, small proteins, and peptides (2D COSY, TOCSY, HSQC, HMBC, NOESY). Thus, this facility has contributed to biomedical research in a unique way over the past 23 years and will continue in this role with this new, very needed instrumentation upgrade.

NSF Awards · FY 2024 · 2024-07

Development research has the potential to improve people’s wellbeing around the globe. Prior research has shown that many of the impediments to economic development are social or institutional in nature. This means that sociology has a critical role to play in advancing research to understand and solve those problems, and is a vital complement to the largely individual and incentive-based approaches of developmental economists. This project is to hold a three-day conference of approximately 120 papers at Johns Hopkins University in fall 2024, in collaboration with the American Sociological Association’s Sociology of Development section, on the theme of "Solving Global Poverty." The conference aims to bring sociological understanding of organizations, systems, governance, and collective action to international development, by bringing together academics and practitioners from the U.S. and around the world. By arrangement with the editors of the journal Sociology of Development, a selection of final papers will be published as a special issue of that journal, and McDonnell and Prasad will be co-editors of the special volume. The conference includes special programs for graduate students, incorporation of practitioners as panel presenters, discussants, and plenary speakers, and other special events. Several practitioners from the World Bank and USAID have agreed to participate and to encourage colleagues to participate. We anticipate more than half of the participants will be from under-represented demographic groups. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2024 · 2024-07

Project Summary / Abstract There are no effective treatments to replace damaged retinal neurons, reflecting a fundamental inability for humans to mount robust regenerative responses within the central nervous system. To address this, it will be critical to understand the diversity of gene regulatory mechanisms that can impede or drive neuron regeneration across vertebrate contexts. Embryonic amniotes have a transitory ability to regenerate retinal neurons from cells of the retinal pigment epithelium (RPE) if supplied with exogenous FGF2 at the time of retinal injury. This mechanism of regeneration can be readily induced at embryonic day 4 (E4) of chicken development, but RPE neural competence is lost by embryonic day 5 (E5). The overarching objective of the proposed research is to profile changes in gene regulation that dispossess RPE cells of their neural competency as they differentiate. Specific aim 1 will interrogate transcription factor regulatory activity within RPE cells across the E4 / E5 developmental window by integrating gene expression analysis, chromatin accessibility profiling, and transcription factor binding assays. Preliminary bulk and single nuclei RNA-seq datasets revealed the acute activation of neural retina transcription factor profiles at both E4 and E5, such as PAX6, ASCL1, and VSX2. In contrast, genes associated with RPE maturity, such as OTX2 and pigmentation genes, were elevated in the E5 RPE independently of retinectomy and FGF2 treatment. Similarly, chromatin accessibility suggested wider dysregulation of OTX2 and related homeobox transcription factor binding sites. During the 1-year F99 phase, OTX2 binding activity will be profiled in intact and FGF2-treated RPE cells at E4 and E5 stages. Additionally, single nuclei RNA-sequencing will capture the heterogeneous transcriptional states of RPE cells during differentiation and FGF2 treatment response at E4 and E5. These results will be integrated to into a model that describes how changes in the RPE gene regulatory landscape culminates in a loss of neural competency. In specific aim 2, the gene regulatory networks present in adult vertebrate models of central nervous system regeneration will be interrogated to inspire novel routes for the induction of mammalian regeneration. This K00 phase will focus on the development of key research skills, including multi-omics data analysis approaches, techniques for spatial transcriptomics and single cell epigenomics, and cross-species genomics / transcriptomics. Up to 4 years will be spent on the K00 phase in an environment directly supportive of these applications. Specific professional development objectives will be concurrently pursued, such as pedagogical development, diversity outreach initiatives, and grant writing. Together, these aims encompass a career development plan that will lead to formation of an independent research program and result in impactful research focused on expanding human central nervous system regenerative capacity.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Injectable therapies for blocking vascular endothelial growth factor (VEGF) have provided impressive initial benefits to most patients with neovascular age-related macular degeneration (nAMD), though some have a suboptimal response. Other vasoactive proteins in addition to VEGF may contribute to the progression of nAMD. An alternative strategy would be to target hypoxia-inducible factor 1 (HIF-1), which is a master regulator of these other vasoactive proteins. We have demonstrated that intraocular delivery of HIF-1 inhibitors is effective in preventing retinal neovascularization (RNV) and choroidal neovascularization (CNV) in a variety of mouse models, where achieving therapeutically relevant drug levels without reaching toxic concentrations is both achievable and key in developing an effective treatment strategy. Another drawback of anti-VEGF therapies has been the disappointing long-term outcomes, as the injections do not eliminate CNV but rather temporarily stop vessel growth and leakage. Further, the short duration of action requires that injections be repeated every 1-2 months. When injections are missed, vessels continue to grow and leak, resulting in permanent retinal damage. One solution is to develop a non-invasive treatment that can be self-administered by patients so that treatment can continue even when patients are unable to return to their retina specialist. However, thus far, it has not been possible to deliver adequate amounts of drug to the retina in large animals (like humans) by topical eye drops. We have discovered an approach for effectively delivering drugs to the posterior segment in large animals, including rabbits and pigs, with once daily topical eye drops. Our unique eye drop-based drug delivery technology provides improved intraocular delivery of both water soluble and water insoluble drugs, including acriflavine. We hypothesize that a new hypotonic gelling eye drop formulation designed to maximize the residence time, intraocular penetration, and drug delivery to the posterior segment with minimal toxicity will be an important step toward the development of a new eye drop-based treatment for nAMD and other ocular diseases with neovascular sequelae. In Specific Aim 1, we will develop and fully characterize new polymer blends for optimal viscosity, shear thinning, gelation rate, and intraocular penetration of acriflavine. In Specific Aim 2, we will test gelling formulations optimized for various key properties versus standard liquid eye drops for maximum tolerated dose and efficacy in preventing CNV in a rat model. In Specific Aim 3, we will evaluate ocular pharmacokinetics and efficacy in rabbits, which are considered a relevant animal model to humans, particularly for characterizing delivery to the posterior segment with topical formulations. We will further evaluate ocular biocompatibility in rabbits, the most commonly used animal model to assess safety of ocular products due to the similarities in eye structure and the sensitivity of the eye to potential toxicity. If these preclinical studies progress as expected, we will be well-positioned for translation of a first-of-its-kind eye drop to the clinic.

NIH Research Projects · FY 2024 · 2024-07

Abstract While over 90% of patients with metastatic prostate cancer respond to systemic therapies that block androgen receptor signaling, nearly all of these patients will eventually relapse and die due to development of acquired therapy resistance. Thus, strategies to prevent acquired therapy resistance are needed. We have identified a rare molecular subtype of prostate cancer with isocitrate dehydrogenase 1 (IDH1) mutation that fails to acquire resistance to therapy. This exploratory proposal will investigate mechanisms by which IDH1-mutant prostate cancer fails to acquire resistance to therapy with a goal to identify novel therapeutic targets to prevent resistance. Molecular consequences of IDH1 mutation in cancer have been previously studied in AML and glioma where it has been shown that IDH1 mutation leads to accumulation of R-2HG, inhibition of 2OG dioxygenases, increased DNA and histone methylation, and marked alteration of epigenetic regulation of gene transcription. However, AML and glioma are different from most solid tumors including prostate cancer because they do not metastasize and have unique treatment paradigms. We will test the hypothesis that IDH1-mutant prostate cancer is restricted in epigenetic control of gene transcription, rendering it less able to adapt to stress such as therapy, and that inhibition of a 2OG-dioxygenase(s) enhances response to therapy. In Aim 1 we will study the largest cohort of patients with IDH1-mutant prostate cancer to date to describe clinical, genomic, and transcriptomic features of this disease. Aim 2 is to develop a transgenic murine model of IDH1-mt prostate cancer to study mechanisms of therapy resistance and sensitivity in vivo. In Aim 3, we will perform a targeted CRISPR-KO screen to identify 2OG-dioxygenases that upon deletion increase DNA methylation and therapy sensitivity in prostate cancer. Together, these aims will provide insight into molecular consequences of IDH1 mutation in prostate cancer and may identify novel drug targets to disable epigenetic plasticity required for acquired resistance to therapy.

NIH Research Projects · FY 2026 · 2024-07

PROJECT SUMMARY Sale of cannabis for recreational use is rapidly becoming legal across states in the US. While cannabis use policies typically contain marketing restrictions, including prohibitions on misleading marketing claims, there is limited research to inform operationalization of these policies, and these policies are rarely enforced. Cannabis products are often labeled based on “species” or cultivar (Indica, Sativa, or Hybrid). Notably, there is limited evidence that different species consistently have different profiles of chemical constituents. Despite this, cannabis is marketed directly to consumers with product labels “Indica”, “Sativa”, and “Hybrid”, and with corresponding marketing claims that the product has sedative (e.g., relaxation, sleepiness) or energizing (e.g., focus, productivity, physical activity) effects. Consumers also report experiencing sedative effects from Indica- labeled cannabis and energizing effects from Sativa-labeled cannabis. This may translate into unsafe use: our prior work has found consumers are more likely to report using Sativa-labeled (vs. Indica-labeled) cannabis before driving or going to work. Despite the potential for public health harm, there is no data demonstrating whether product perceptions, use expectancies, and subjective and objective acute effects of use vary by label or marketing claim. Research from other consumer domains, including tobacco, indicates that labeling and marketing can have powerful effects on product perceptions, use behavior, and use experience. Marketing and labeling for cannabis could similarly foster inaccurate product perceptions (e.g., regarding harm), expectancy effects (e.g., anticipating a product to increase focus) and lead to unsafe use (e.g., while driving). This undermines initiatives to promote safe use of cannabis and risks public health. To date, controlled research has not systematically evaluated how product labeling (Indica, Sativa, Hybrid) or associated marketing claims (sedative vs. energizing) affect use behavior, risk perception, or acute drug effects. The proposed research uses a large-scale content analysis of cannabis labeling and marketing (Aim 1), a randomized online experiment (Aim 2), and a placebo-controlled randomized laboratory experiment (Aim 3) to document the scope and effects of cannabis labeling (Indica, Sativa, Hybrid) and marketing claims (sedative, energizing). Ultimately, this work will provide evidence to inform cannabis marketing and labeling policies, which are necessary to protect public health.

NIH Research Projects · FY 2025 · 2024-07

The mission of the Johns Hopkins University (JHU) Institute for Clinical and Translational Research (ICTR) Predoctoral Clinical Research Training Program (T32) is to provide an integrated set of training opportunities focused on the core principles of translational science. We will build on our success of training more than 150 predoctoral students to date by focusing the training experience on (1) specific skills for competency in research (2) individual mentored project development and (3) communication of research results to nonscientist audiences. A core program goal is to promote multidisciplinary research collaborations in translational science. We have a track record of training scholars who are enrolled in a variety of doctoral programs including medicine, nursing, public health. We will continue to advance this goal by fostering collaboration across scientific disciplines. Our history of successful recruitment and selection of trainees from multiple participating institutions allows us to build on our efforts to enroll trainees from across disciplines. We will use existing strategies to recruit trainees in medicine, nursing, dentistry, and public health with a variety of scientific interests and perspectives in translational science. We (1) Teach core principles in clinical and translational research, ensuring trainees have the knowledge and skills to succeed in their scientific pursuits and build an identity as translational researchers; (2) Provide a supportive and collaborative culture for trainees; (3) Create a community of clinical and translational science trainees, building networks and collaborations among trainees (and mentors), driving innovation; and (4) Encourage the development of team science skills. Trainees will recognize the importance of working collaboratively, an increasingly critical research skill. Our integrated program offers a multidisciplinary mentoring team, didactic training, competency-based interactive workshops, core ICTR resources, peer-support and resource-sharing designed to stimulate lifelong engagement in a clinical and translational research career. The program offers trainees intensive and hand-on learning experiences that prepare them to be exemplary clinical and translational scientists in any discipline, specialty or subspecialty. Our multidisciplinary pool of trainees will emerge equipped with the knowledge, skills, and abilities to advance diagnostics, therapeutics, clinical interventions, and behavioral modifications to improve health as they develop into independent researchers. We will perform continuous program evaluation with multiple metrics. We evaluate the program itself (faculty, courses, seminars) and track enrollment, retention, and the range of scientific disciplines of trainees; and conduct course and mentoring evaluations. Like the consortium of CTSA academic medical institutions across the country, we will report on trainee performance and satisfaction, and mentor satisfaction. We will also measure, evaluate, and disseminate the impact of translational research from our program. The JHU ICTR T32 will support 10 predoctoral trainees per year for a minimum of 12 months.

NIH Research Projects · FY 2024 · 2024-06

Project Summary Peripheral neuropathies are major neurological complications of multiple chemotherapy drugs causing significant morbidity affecting quality of life and potentially altering life-saving chemotherapy regimens. Many chemotherapy drugs with diverse mechanisms of actions cause axonal degeneration and the underlying mechanisms that lead to distal axonal degeneration, a common feature of most peripheral neuropathies, are poorly understood. Furthermore, currently there are no therapies aimed at preventing, reversing, or slowing the progression of peripheral neuropathies that cause chronic neuropathic pain, sensory loss, and weakness. In this exploratory R21 grant we will test the overall hypothesis that MAP4K4 inhibition protects peripheral sensory neurons against toxicity of several chemotherapy drugs and prevents chemotherapy- induced peripheral neuropathy (CIPN). This hypothesis is built upon unpublished preliminary data that we generated through an unbiased kinase inhibitor screen we performed to prevent chemotherapy induced neurotoxicity. In these preliminary studies we found that PF-06260933 dihydrochloride (PF062), a MAP4K4 inhibitor, provided robust protection against neurotoxicity of Paclitaxel (PTX), and Cisplatin (CDDP) and Bortezomib (BTZ) in vitro. These studies suggest that axon degeneration cascades initiated by three chemotherapy drugs with different mechanisms of action (paclitaxel, cisplatin, and bortezomib) converge on MAP4K4 and that inhibitors of MAP4K4 can be used to prevent neurotoxicity of different chemotherapy drugs. We propose to test this hypothesis further by validating the neuroprotective effects of MAP4K4 inhibitors from different chemical classes as well as using genetic knockdown experiments. Furthermore, we will test the role of MAP4K4 inhibition in vivo. Completion of these studies will give us a better understanding of the role of MAP4K4 in distal axonal degeneration in CIPN and help further explore a novel therapeutic target that can be taken to clinical studies in a timely manner.

NIH Research Projects · FY 2025 · 2024-06

Project Summary/Abstract Cell migration is essential for many physiological processes including embryonic development, wound healing, and immune responses. Cells achieve efficient directional migration through two major steps: front-and-back polarization and re-arrangement of interactions between the cell and extracellular matrix (ECM). Cells adhere to the ECM using actin-based multiprotein complexes called focal adhesions (FAs) with which the cells exert forces to push or pull themselves to migrate, while long-term polarity is maintained by another cytoskeletal component, microtubules (MTs). Therefore, efficient cell polarity and migration are achieved by coordinated regulation of actin, MTs, and FAs. However, it remains unknown if there is a central regulator that orchestrates these seemingly distinct subcellular organizations. Our lab has recently found that cells lacking α-tubulin acetyltransferase 1 (αTAT1), the sole mediator of MT acetylation, display defects in FAs as well as in front-and- back polarity. In addition, preliminary data also showed that cells with an αTAT1 knockout (KO) migrate faster compared to control cells during random migration assays. We hypothesize that MT acetylation is a master regulator of cell migration by dynamically remodeling molecular constituents and their activity at the F-actin and MT cytoskeletons and FAs. To test this hypothesis, I will combine live-cell, time-lapse fluorescence microscopy with pharmacological and genetic perturbations to elucidate how acetylated MT affects single and collective migration (Aim 1), and to reveal molecular mechanisms regulating the front-and-back polarity (Aim 2) and FAs dynamics (Aim 3) in a MT acetylation dependent manner. In Aim 1, I will characterize the role of acetylated MTs in cell migration by performing single and collective migration assays under different levels of MT acetylation and quantifying migration characteristics such as velocity, directionality, and wound closure rate. Actin polymerization in the front defines the leading edge of a migrating cell; this reaction is regulated by spatially restricted activities of Rho family GTPases. Therefore, to investigate the role of acetylated MT in cell polarization (Aim 2), I will use time-lapse fluorescence microscopy to measure the dynamics of MTs and F-actins and quantify the activity of Rho GTPases using FRET sensors. In Aim 3, I will elucidate how MT acetylation modulates biophysical properties of FAs in migrating cells by examining the expression level and localization of FA proteins and measuring FA dynamics using optogenetics and live-cell fluorescence imaging. In addition, I will quantify the force exerted using traction force microscopy in cells with three different conditions where the extent of MT acetylation is varied (normal, none, or elevated). Together this proposal will illuminate a role of acetylated MTs as a regulator of directed cell migration by concertedly regulating front-and-back polarity and FAs dynamics.

NIH Research Projects · FY 2026 · 2024-06

PROJECT SUMMARY Antimicrobial resistance (AMR) in Neisseria gonorrhoeae (NG) is in the top tier of AMR threats as defined by WHO. NG is responsible for the second most prevalent sexually transmitted infection worldwide, which can cause long-term health consequences. Unfortunately, NG has rapidly developed resistance to all first-line antimicrobials previously recommended for the treatment of gonorrhea, including the last viable antimicrobial, ceftriaxone. As AMR continues to evolve, there is an urgent need for personalized treatment approaches that can spare the use of ceftriaxone to reduce the chances of NG becoming resistant to this antimicrobial. However, this requires clinicians to know drug resistance or susceptibility quickly enough to inform prescription decisions. The Centers for Disease Control and Prevention (CDC) periodically publishes STD treatment guidelines to help clinicians, which are informed by susceptibility data generated by the national CDC Gonococcal Isolate Surveillance Project (GISP). However, determining AMR requires prolonged (24-48 hours) microbiological cultivation in sophisticated laboratory facilities, which has been supplanted by nucleic acid amplification tests (NAAT) for diagnosis of NG infections. The widespread adoption of NAAT has created a critical void in AMR testing, leading to a loss of capability to perform culture of NG in most testing clinics. We propose to develop a rapid AST platform for NG, capable of testing against multiple antimicrobial conditions at scale directly from clinical specimens. Our proposed method involves measuring the earliest transcriptional responses to antimicrobials for rapid AST. We and others have shown that measuring mRNA expression levels following a 10-minute antimicrobial exposure can predict susceptibility well before phenotypic changes in growth are observable. This forms the basis of our proposed single-cell mRNA-enabled AST (sc-mRNA-AST) for NG. To achieve sc-mRNA-AST for NG, our platform will use droplet microfluidics to isolate and analyze individual NG cells from clinical specimens with heterogeneous bacterial flora. We will first identify the most susceptibility- informative transcripts agnostic to mechanisms of resistance against each of 3 NG-relevant antimicrobials. We will use ultrasensitive single-molecule fluorescence spectroscopy to quantify mRNA molecules from individual cells without the need for amplification. Additionally, we will incorporate an assembly-line-like, cascaded microfluidic design into our platform, allowing for numerous sc-mRNA-AST assays against clinically relevant antimicrobial conditions. With an additional 2 minutes per condition, our platform can test 15 antimicrobial conditions to determine susceptibility and minimal inhibition concentrations (MICs) for 3 antimicrobials within 1 hour, directly from clinical urogenital samples. To evaluate the performance of our platform, we will use contrived samples with NG isolates and prospectively collected clinical samples. Successful development of sc-mRNA- AST for NG will lead to personalized treatment to spare ceftriaxone use, and improved AMR surveillance to inform antimicrobial stewardship and public health policy.

NIH Research Projects · FY 2024 · 2024-06

PROJECT SUMMARY/ABSTRACT People living with HIV experience neurologic dysfunction, including encephalitis, depression, anxiety, and cognitive deficits. Chronic inflammation is a major hallmark of HIV disease today and contributes to the development of neurologic dysfunction in up to 50% of people living with the virus. These neurologic deficits result in poorer daily functioning, decreased quality of life, and have become more prevalent due to the survival benefits of antiretroviral therapy (ART). Thus, chronic inflammation and its consequent effect on neurologic health represents a major public health issue. Microglia are an integral underlying mediator of this chronic inflammation through production of inflammatory cytokines, chemokines, and antiviral responses. Together, these components promote neuroinflammation that persists despite ART and contributes to neurologic impairment through demyelination, pruning, and destruction of neurons. There is a pressing need to develop adjunct therapies to limit deleterious effects on neurologic health. Cannabinoids are emerging as an important modulator of inflammation. Phytocannabinoids, including cannabidiol (CBD), are known to modulate inflammation through activation of canonical and extended endocannabinoid system receptors. While canonical endocannabinoid receptors (CB1 and CB2) are the most well studied, less studied extended endocannabinoid receptors (TRPV2 and PPAR-a) also possess immune modulating functions. The research goal of this F31 proposal is to evaluate expression, function, and immunomodulatory potential of endocannabinoid receptors, and CBD, in microglia in the context of HIV. Our preliminary data determine that macrophage/microglia express both canonical and extended endocannabinoid receptors. Of note, they express TRPV2 and PPAR-a more than any other brain cell type, including neurons, astrocytes, endothelial cells, and pericytes. We determined that endocannabinoid receptor pharmacologic agonists modulate myeloid cell inflammation following exposure to potent bacterial moiety, lipopolysaccharide, an example of a strong inflammatory agent. Together, these preliminary data suggest endocannabinoid receptors are present on myeloid cells and have the capacity to modulate chronic, myeloid derived inflammation. We hypothesize that 1) CB1, CB2, TRPV2, and PPAR-a are present on macrophage/microglia, are selectively impacted by HIV infection, and that they 2) mechanistically contribute to modulation of HIV inflammatory responses. We will systematically test this hypothesis by evaluating the impact of HIV on expression and function of these receptors. We will also use CBD and agonists to evaluate the mechanistic contribution of receptors to modulating microglial inflammation. Successful completion of these research goals along with the proposed mentorship and training provided through this F31 award will propel me forward in accomplishing my long-term goal of becoming a public health research scientist working at the intersection of HIV and substance use.

NSF Awards · FY 2024 · 2024-06

The equatorial electrojet (EEJ), an intensified electric current flowing eastward in the ionosphere during daytime hours. The work will investigate whether the EEJ weakens or strengthens during substorms. Successful completion of this project will not only resolve this controversy but will also significantly improve our overall understanding of the global current circuit. Understanding how the upper atmosphere at the auroral region couples with that at low latitudes is a crucial step for ensuring space weather awareness and mitigating the harmful effects of geomagnetic perturbations on technology, such as power grids, satellites, and communications. The investigators will address the questions related to the spatiotemporal characteristics of the EEJ and the interplanetary magnetic field (IMF) conditions during substorms. These are: (i.) How does the EEJ response to substorms vary with longitude, local time, season, and solar flux? (ii.) How do substorm duration, intensity, onset time, and IMF conditions affect the EEJ response? The methodology hinges on observational data and robust statistical analysis, leveraging NSF-supported projects, the SuperMAG network, and the Jicamarca Radio Observatory (JRO). Based on four well-known substorm lists, the research team intends to examine events during geomagnetic quiet periods and quasi-steady IMF conditions. These stringent criteria allow them to isolate the sole substorm effect and exclude unconnected factors like IMF variations known to impact the EEJ. The resulting magnetic field perturbations will be assessed using a well-established approach based on a pair of magnetometers, one positioned within ±3° of the dip equator and the other at an offset location (6°-10°). They will assess the related electric field variations via vertical ion drift measurements from the incoherent and coherent scatter radars at the JRO. The success of our proposal is sustained by the wealth of data and the expertise of the team. The investigation will provide new insights into the influence of EEJ response to substorms, thus providing new perspectives on how the electric fields penetrate from high to low latitudes, improving their understanding of the magnetosphere-ionosphere current system, and supplying essential parameters for future simulations. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2024 · 2024-06

PROJECT SUMMARY We are requesting a Leica STELLARIS 8 FALCON laser scanning confocal microscope for the microscopy facility in the Department of Molecular Microbiology and Immunology at the Johns Hopkins Bloomberg School of Public Health, which is part of the Johns Hopkins Medical Campus. A unique feature of our facility is the ability to perform live cell imaging experiments with biological agents that require Biosafety level 2 (BSL-2) containment, including human pathogens, such as Influenza A virus, Cryptococcus neoformans, Borrelia burgdorferi, Listeria monocytogenes, Plasmodium falciparum and Toxoplasma gondii. The proposed instrument will represent the first confocal microscope for this facility and within the current space restrictions will be singularly capable of supporting the various advanced confocal microscopy needs of our users. Notable features that convinced us during testing are its Wight Light Laser, translating into 350 laser lines, its imaging speed and the capacity to detect up to five different fluorophores simultaneously, while offering the gentle imaging conditions needed for live microscopy at confocal resolution. TauSense and FALCON enable the harnessing of fluorescence lifetime- based information and with this provide additional image quality, the ability to separate overlapping fluorophores and state-of-the-art measurements of protein interactions via FLIM-FRET. Altogether, the addition of this instrument to our facility will significantly enhance NIH-funded projects of our users and greatly contribute to the infectious disease focus and other key research areas of our institution.

NIH Research Projects · FY 2026 · 2024-06

Project Summary/Abstract Urine drug testing (UDT) is an integral component of opioid use disorder (OUD) treatment, and is often required by payers for patients treated with medications for OUD (MOUD). These regulations exist despite a lack of evidence-based guidelines. There is no consensus about the optimal UDT frequency, what drugs to test for, when to use different types of tests (e.g., presumptive versus definitive), and how to adapt care when patients test positive for non-prescribed substances. Inconsistent practices have important public health and clinical implications, as they can contribute to either missed opportunities to improve care or to the misallocation of clinical resources. The goal of this grant is to identify variation in real-world use of UDT that can be modified by clinical guidelines and policy change, and to examine the patient- and clinician-level factors that influence the application of UDT, and their relationship with processes of care. Aim 1 will characterize distinct trajectories of UDT use during episodes of OUD treatment nationally, and across patient characteristics, clinicians, payers, and states. The sample will comprise continuously enrolled patients with diagnosed OUD from 2015-2022 from a national, multi-payer database that tracks patients across disparate data systems and includes service claims data linked to laboratory results. We will examine the correlates of testing frequency and type across different groups of interest. Aim 2 will identify distinct trajectories of drug positivity during OUD treatment episodes and identify variation across groups. We will identify the prevalence of patients testing positive for common non- prescribed drugs (e.g., opioids, cocaine) at the beginning of treatment episodes and the persistence of positivity in the treatment episode. Using latent class analysis, we will group patients into subtypes related to patterns of drug positivity (e.g., persistently abstinent, transitioning to abstinence, persistently positive). Aim 3 will estimate the association between trajectories of UDT use, substance positivity, and process of care measures in the episode (e.g., retention, MOUD adherence, hospitalization). Using Aims 1-2 variables, we hypothesize that greater UDT use will not generally lead to changes in process of care outcomes and that patients who are persistently positive for substances will have worse process of care outcomes. Aim 4 will examine perspectives of clinicians to identify perceptions and practices related to UDT use. We will survey a national sample of clinicians treating OUD recruited from a drug testing laboratory. They will be asked about their knowledge, attitudes, and practices related to testing. From this group, we will conduct qualitative interviews with 30 clinicians who vary in their testing practices to learn more about potential opportunities to inform decision- making surrounding UDT in clinical practice. Overall, this study will characterize the variation in UDT use in real- world clinical practice and help identify the potential scope for UDT to positively change processes of care for patients. The findings will lay the foundation for developing pragmatic clinical trials, informing policy decisions, and refining clinical practice.

NIH Research Projects · FY 2025 · 2024-06

PROJECT SUMMARY Anaphylaxis is a systemic, potentially fatal, immediate hypersensitivity reaction (IHR) that can be triggered by foods, drugs, vaccines, singing insect venom, exercise, or even idiopathically. Accurate diagnosis is essential in both the acute setting, when medical intervention can be life-saving, and during subsequent outpatient evaluation, when management depends on confirming the diagnosis and identifying the triggering substance. The diagnosis of anaphylaxis is based on symptoms, which is problematic due to the wide variability of clinical presentations and substantial overlap of allergic symptoms with other disease states. These diagnostic complexities and lack of reliable biomarkers for IHRs contribute to the misdiagnosis and subsequent mismanagement of food and drug allergies. Current known biomarkers for systemic allergic reactions are lacking in that none are both sensitive and specific. Therefore, there is a sizable unmet need to discover novel biomarkers of IHRs that are both highly-sensitive and specific for all contexts, mechanisms, and severities of anaphylactic reactions. This proposal’s central objective is to characterize and quantify circulating extracellular vesicles (EVs) as novel biomarkers of acute IHRs using patient samples obtained from ongoing clinical trials. Aim 1 will characterize the quantities, surface marker expression, and contents of plasma cell-specific EVs in adults after acute IgE-mediated IHRs to foods, and compare findings to patients’ baseline. Aim 2 will similarly characterize EVs in adults after acute non-IgE-mediated IHRs to drugs. Aim 3 will create a biobank of patient samples for analysis of alternative biomarkers using cutting edge technologies. This highly feasible and valuable clinical research project is expected to identify novel biomarkers that will aid in the accurate diagnosis of anaphylaxis as well as provide new insight into the mechanisms of certain drug IHRs. This application is in response to the program announcement for Exploratory and Developmental Research Grant Program for NIAID K-award Recipients. This award will provide crucial support and resources for the PI as she transitions to research independence.

NIH Research Projects · FY 2025 · 2024-06

PROJECT SUMMARY Metastasis is the primary cause of cancer mortality, yet few breast cancer drugs effectively inhibit metastasis. Breast cancer cells use collective migration to remodel and align surrounding extracellular matrix (ECM) fibrils, which facilitates invasion. Aligned tumor stroma topography can induce cluster budding and dissemination of breast cancer cells. The goal of this project is to identify chemotherapeutic drugs using engineered biomimetic tumor invasion models and to evaluate therapeutic feasibility of inhibiting the target genes involved in breast cancer dissemination. To achieve this goal, we developed a quasi-3D nanotopographically patterned substrate and are incorporating it into a nanopatterned impedance electrode array (nanoIEA) to quantify collective cell migration and proliferation in real-time at high-throughput. We are validating a 3D aligned collagen fiber hydrogel model with control over fiber alignment that recapitulates the fiber dimension and orientation of in vivo breast tumor stroma. These models markedly promote breast cancer cluster dissemination and increase its resistance to chemotherapy. In our preliminary study, we have identified differentially expressed genes via RNA-seq between ‘disseminated tumor cell clusters’ and ‘non-disseminated tumor cells’ using the quasi-3D model. We will pursue three aims that leverage our expertise in cancer molecular biology/genomics (Ahn), tissue engineering (Kim), machine learning (ML)-based image analysis (Lee), cancer organoids/metastasis (Ewald), and pharmacology/drug development (Liu). Human breast cancer patient-derived xenograft (PDX) cell clusters/organoids will be investigated in this project. In Aim 1, we will evaluate effects of the following drugs on collective cell migration and on growth using the nanoIEA: [a] the 23 oncology drugs (out of 147 drugs we tested) which most significantly inhibited the viability of breast cancer cells in the quasi-3D model, [b] the 73 non- oncology drugs which inhibited the viability of 22 breast cancer cell lines by at least 4-fold in conventional 2D culture, and [c] the 95 inhibitors of target genes (CYP1A1, CYP1A2, CYP1B1). In Aim 2, we will characterize phenotypic responses of breast cancer cells/organoids to the identified drug candidates from Aim 1 using live cell microscopy and ML analyses. Phenotypic changes (e.g., motility, morphology) will be quantified to contrast subpopulations (non-invasive vs. invasive) and drug-treated cells vs. untreated. In Aim 3, we will evaluate therapeutic feasibility of regulating the target genes to inhibit cancer invasion. We will determine expressions of target genes at protein levels in PDX organoids, then correlate these with organoid invasiveness in the 3D model. We will then determine how inhibition of the target genes influences chemosensitivity of PDX organoids and suppresses their invasiveness. This project will increase our understanding of the mechanisms of topography- induced breast cancer dissemination and establish our tumor ECM-mimetics, nanoIEA, and ML imaging analysis as a preclinical cancer invasion model/assay to characterize heterogenous cell populations with different metastatic phenotypes and to identify chemotherapeutic agents that directly inhibit breast cancer invasion.

NIH Research Projects · FY 2025 · 2024-06

Cancer-associated fibroblasts (CAFs) are abundant components of the tumor microenvironment in the skin of early-stage mycosis fungoides (MF) patients. Evidence suggests CAFs promote the progression of early-stage MF to late-stage MF by suppressing the tumor surveillance function of the immune system. Currently, little is known about the CAFs responsible for immune evasion and disease progression in MF or how this differs by patient demographics or stage of disease. As CAFs are heterogeneous cells, investigating CAFs requires novel molecular techniques that account for the diversity of cell populations and can determine the location of critical subtypes. The Research Training Plan will use the novel molecular techniques of single-cell RNA sequencing (AIM 1) and spatial RNA transcriptomics (AIM 2) to uncover the subtypes and location of CAFs responsible for immune evasion and MF progression from a diverse patient population. In Aim 1, Dr. Johnson will work with mentor Dr. Winston Timp and advisor Dr. Ron Sweren to obtain patient samples, dissociate whole skin tissue, and perform single-cell RNA sequencing on patients from diverse stages and demographics in the K99 phase. Dr. Johnson will train with advisors, Drs. Elana Fertig and Stephanie Hicks to develop computational analysis and apply statistical models to identify subtypes of CAFs based on gene expression. Advisor Dr. Jaroslaw Jedrych will provide guidance with immunohistochemistry verification of gene biomarkers. The R00 phase will include the analysis to identify biomarkers relevant to MF disease diagnosis and prognosis in this diverse patient population. In Aim 2, Dr. Johnson will work with mentor Dr. Winston Timp and collaborator Dr. Elana Fertig to execute spatial RNA sequencing techniques and acquire computational analysis to integrate single-cell RNA sequencing data with spatial RNA sequencing data to locate the subtypes of CAFs identified in AIM 1 in the K99 period. The R00 period will further explore all cells within the tumor microenvironment to uncover candidate cytokines and receptors responsible for crosstalk pathways between the MF tumor cells and the CAF subtypes of interest. Dr. Johnson has had some prior training in single-cell RNA sequencing and analysis and expertise in skin biology. The Career Development Plan is tailored to enable Dr. Johnson to develop computational analytical skills and refine her statistical analysis of single-cell and spatial data while gaining valuable background skills through coursework. Mentor Dr. Winston Timp is a leading expert in sequencing and genomics. Advisors Dr. Stephanie Hicks, an expert in single-cell statistical analysis, and Dr. Elana Fertig, an innovator in computational analysis of integrative transcriptomics, offer complementary expertise. The JHU Department of Dermatology and Biomedical Engineering environment is an outstanding collaborative setting with excellent core facilities and biorepositories infrastructure. In summary, the strong mentoring environment and training plan are anticipated to prepare Dr. Johnson to launch her independent career. The proposed studies will offer mechanistic insights into the pathogenesis of MF progression and immune evasion in a diverse patient population.

NIH Research Projects · FY 2026 · 2024-06

PROJECT SUMMARY Infection-induced B cell activation occurs in the context of complex innate immune responses, including the elaboration of cytokines, and the remodeling of secondary lymph tissues draining sites of infection. Our long-term objective is to determine how optimal protective humoral immunity to infections is induced and maintained. Exploring infection-induced innate stimuli that modulate adaptive immunity, we showed that infection-induced and TLR-agonist adjuvanted s.c. immunization provide required B cell intrinsic TLR signaling via both adaptors (MyD88 and TRIF) for extrafollicular responses, as deletion of both adaptors abrogated EF responses. While B cell intrinsic MyD88 stimulation drives B cell proliferation, TRIF signaling does not, indicating distinct yet unknown mechanisms of TRIF-mediated support for EF formation. When and where, during B cell activation, these innate signals regulate B cell fate is unknown. Recently we identified an early induced, post proliferative, influenza specific B cell population with a unique phenotype and transcript profile that might represent an activation intermediate of EF. The objectives for this project are to test the hypothesis that early in influenza infection B cell intrinsic TRIF/TLR3 and MyD88-mediated signals support EF differentiation via two distinct differentiation paths, each differently affecting extrafollicular antibody quality, acting on antigen-stimulated, post-proliferative B cells. To achieve our objectives, we will test in Specific Aim 1 the hypothesis that B cell intrinsic TLR3-signals support effective plasmablast differentiation by enhancing B cell responsiveness to IL2, while MyD88 principally drives proliferation followed by terminal differentiation, differentially affecting antibody quality. In Specific Aim 2 we will determine the extent to which the population of transcriptionally distinct, antigen-experienced, mostly non-switched and post-proliferative B cells that appear in the draining LN at 5 dpi with influenza virus, represent an intermediary, quiescent stage in B cell activation to EF, extrafollicular memory B cells and/or GC development and determine the impact of TLR signaling on their development and fate. Successful completion of the work would provide significant conceptual advances to understanding early B cell activation and extrafollicular memory B and plasma cell response induction. It would also identify a novel role for TLR3 in support of EF development and clarify the function of TLRs on B cell responses. Collectively these advances would identify new, critical junctures in B cell activation, regulated by innate immune signals for potential exploitation in vaccine design.

NIH Research Projects · FY 2024 · 2024-06

Abstract The DASH (Dietary Approaches to Stop Hypertension) eating pattern has been shown to reduce systolic and diastolic blood pressure by clinically meaningful levels (10.7 and 4.7 mm Hg, respectively) among adults with hypertension. Accordingly, national treatment guidelines recommend the DASH as a first-line treatment for the 116 million US adults with hypertension. However, DASH adherence among U.S. adults with hypertension remains exceedingly low. A major barrier to DASH uptake is poor accessibility to and high cost of healthful foods. Approximately 38% of U.S. census tracts have limited access to high-quality foods; these communities generally lack supermarkets or large grocery stores, which facilitate both availability and affordable food pricing. Healthful food access is especially constrained in lower-income and predominately racial/ethnic minority neighborhoods. To best address the considerable population health challenge of hypertension, we need innovative approaches to expand equitable access to the DASH eating pattern. Members of our team are currently conducting Nourish (R01HL146768), a digital behavioral health intervention delivered via a smartphone application (app), to improve adherence to DASH among adults with hypertension. The intervention components include DASH education and skills training, self-monitoring, personalized feedback, goal setting, and responsive health coaching. We expect that the Nourish intervention, if successful, can be scaled and disseminated to millions of adults. However, access challenges remain; without additional intervention, a large proportion of U.S. adults will still have limited access to high-quality foods--a critical barrier to adopting the DASH eating pattern. In the current proposal, we will leverage the rapidly mounting availability of grocery delivery services to deliver Access Nourish, an intervention that will provide tailored grocery lists and at-home grocery delivery options through a commercially available smartphone app. We will test the feasibility of Access Nourish among n=65 patients with hypertension living in low access census tracts, as defined by the USDA. We will deliver Access Nourish in addition to the existing intervention components of Nourish, described above, to provide a comprehensive intervention package that targets behavioral mechanisms to increase DASH adherence and improves food access. Feasibility will be assessed via participant accrual; participant retention; and other indicators of feasibility, including acceptability, demand, implementation, practicality, and adaptation of the intervention. We will also conduct qualitative interviews to elicit barriers and facilitators to using an online grocery delivery app. We believe this approach will yield meaningful information regarding the utility of these tools in improving access and adherence to effective hypertension management interventions. Subject to future efficacy testing, this intervention can be scaled to enhance dietary interventions that are needed by millions of Americans who lack access to high-quality food, yet require dietary modifications to control their high blood pressure and related comorbidities.

NIH Research Projects · FY 2025 · 2024-06

Project Summary Malaria continues to have a huge toll on morbidity and mortality in the world, responsible for over 600,000 deaths annually. The transmission stages are bottlenecks for the parasite and thus critical stages for intervention, particularly when aiming for elimination. Using the rodent model Plasmodium yoelli, we recently found that salivary gland sporozoite load of a biting mosquito strongly correlates with infection likelihood, with highly infected mosquitoes being 7.5 times more likely to initiate an infection. Importantly, the likelihood of achieving secondary infections rapidly rises at salivary gland sporozoite densities >10,000 that represent a small proportion of field- caught mosquitoes. These data challenge the assumption that all infected mosquitoes are equal, a belief that underlies epidemiological models of transmission and impacts malaria elimination strategies. In this proposal we will determine the mechanisms underlying the increased infectious potential of highly infected mosquitoes, comparing the quantity of sporozoites inoculated by mosquitoes harboring high and low numbers of sporozoites, as well as the quality of sporozoites in these two groups. In Aim 1 we will determine the inocula associated with low- and high-infected mosquitoes using both rodent malaria parasites and the human parasite P. falciparum. In Aim 2, we will compare sporozoites from mosquitoes with low and high infections using infection assays, intravital imaging, and single-cell RNA-Seq analysis. Overall these experiments will fill crucial knowledge gaps on parasite transmission probability that will greatly improve our understanding of parasite biology and epidemiology.