Johns Hopkins University

NIH Research Projects · FY 2025 · 2022-08

Project summary/abstract Infection with human papilloma virus (HPV) causes endemic occurrence of cervical, head and neck and anal cancers. Radiotherapy is the cornerstone treatment for these tumors. However, clinical outcomes in patients with locally advanced disease remain poor due to high recurrence rates. Thus, there is a critical need to improve the efficacy of radiation in these tumors. Epigenetic signaling pathways play a crucial role in DNA damage response providing suitable clinical targets for improving the efficacy of radiotherapy. We demonstrate that expression of the HPV proteins E6 and E7 alters epigenetic signaling in cancer cells. We hypothesize that these epigenomic alterations cause tumor dependence on alternative epigenetic pathways to cope with DNA damage. Using a CRISPR/Cas9 screen we discovered that HPV-positive cancer cells selectively depend on the chromatin modifier NSL1 and the E3 ubiquitin ligase RNF168 to survive radiation. The goal of this proposal is to determine the mechanisms of DNA damage response (DDR) regulated by these pathways in the setting of HPV-induced epigenetic changes. Our work will establish a novel concept of tumor-targeted therapy by harnessing HPV- induced epigenomic alterations to allow specific targeting of cancer cells while sparing healthy tissues. The work proposed here will be conducted under the mentorship of Dr. Dennis Hallahan, a leader clinical and experimental radiation oncology. The candidate is an MD/PhD with training in clinical radiation oncology who seeks further training in basic research. His long-term goal is to establish an independent research laboratory studying the role of epigenetics in DDR. It is anticipated that the project will result in impactful contributions to the fields of DDR and Radiation Oncology and prepare the candidate for a career as an independent investigator.

NIH Research Projects · FY 2025 · 2022-08

This K01 Mentored Research Scientist Development Award application will facilitate the Principal Investigator’s career development as a leading, productive, independent researcher conducting innovative implementation science to maximize the real-world impact of human immunodeficiency virus prevention interventions in low-resource settings. The proposed training plan and research implementation in Zambia will allow her to meet her training objectives, developing the critical knowledge, skills, and competencies required to transition into an independent researcher. She will: 1. Gain expertise in pre-exposure prophylaxis (PrEP) and prevention interventions, 2. Develop skills in evidence-based, stakeholder-informed implementation strategy specification to optimize population-level impact of novel interventions, 3. Develop statistical expertise in discrete choice experiments and latent class analysis, 4. Establish capacity in internet-based recruitment and data collection in a low-resource setting, 5. Understand and measure developmental differences, including cognitive and affective development, among adolescents and young adults and their associations with HIV prevention. Her well-established mentoring team will support her successful achievement of her training, research, and career development goals. Her primary mentor is an international expert in HIV prevention. She has five co-mentors offering expertise in Cabotegravir long-acting injectable prevention; implementation science with latent class analysis expertise; development, execution, and analysis of discrete choice experiments; adolescent and young adult development; and human immunodeficiency virus research in Zambia. Her Scientific Advisor is an expert in internet-based research in low-income settings. Cabotegravir long-acting injectable prevention was found superior to daily oral PrEP in clinical trials and received United States Food and Drug Administration approval in December 2021 with international regulatory approvals following. Cabotegravir long-acting injectable prevention has the potential to be effective among people who have found oral PrEP ineffective, including adolescents and young adults in sub-Saharan Africa who remain susceptible to new infections. The proposed research utilizes implementation science to establish evidence needed to optimize Cabotegravir long-acting injectable prevention implementation among adolescents and young adults in Zambia, minimizing the time from proven intervention efficacy to population impact: Aim 1. Assess the fidelity and sustainability of implementation strategies for two related interventions: oral PrEP and injectable contraception, to inform Cabotegravir long-acting injectable prevention implementation using participatory process mapping; Aim 2. Identify preferences for Cabotegravir long-acting injectable prevention implementation and heterogeneity among adolescents and young adults and healthcare workers using discrete choice experiments and latent class analysis with internet-based recruitment; Aim 3. Specify Cabotegravir long-acting injectable prevention implementation strategies to support use and persistence. A future National Institutes of Health grant will evaluate implementation.

NIH Research Projects · FY 2025 · 2022-08

PROJECT SUMMARY Danetta Sloan is an Assistant Scientist, in the Department of Health, Behavior and Society with education and professional experience as a homecare hospice Social Worker, addressing the needs of patients and families experiencing end stage Alzheimer’s disease. Her long-term goal is to become a leader in community-based research to develop, translate, and implement evidenced-based, real world interventions to reduce inequities in dementia care and access for Aging Black Americans. This award will ensure Dr. Sloan has the knowledge, skills, and experience to develop, implement and test interventions that build individual and community capacity to improve early diagnosis, access and quality of care to Black Americans. Dr. Sloan has convened an outstanding mentorship team with expertise to help develop her career development and provide training in the relevant research methods to adapt, design cultural appropriate, communication skill building intervention, to implement and test in the Black Faith community. Despite the growth in dementia information and dissemination, Black American communities still experience late diagnosis, and suboptimal care, engaging with dementia care providers later in the disease without adequate knowledge to participate in goal of care conversations. With no cure for Alzheimer’s Disease Related Dementias (ADRD), morbidity and mortality remain high and Black Americans are disproportionately impacted and more likely to have ADRD and be unaware. This proposal addresses goals of the National Plan to Address Alzheimer’s Disease to 1) optimize care quality and efficiency and 2) enhance public awareness and engagement. This proposal will equip individuals with skills to identify signs and symptoms, (increase health literacy in dementia) and goal of care communication training pre-clinical diagnosis. The outcome of the intervention in Aim 3, will afford those who experience dementia the opportunity to self-advocate, participate in research, and engage in quality of life decision-making and advance care planning. Feasibility, acceptability, comprehension and satisfaction, with the intervention and self-efficacy to engage in a goal of care conversation will be assessed.

NIH Research Projects · FY 2024 · 2022-08

PROJECT SUMMARY Pulmonary arterial hypertension (PAH) remains a fatal diagnosis despite available therapies. PAH is characterized by extensive remodeling of the pulmonary vasculature involving the formation of vaso-occlusive lesions and a thickened medial layer of the vascular wall, both of which contain pulmonary arterial smooth muscle cells (PASMCs). It has been demonstrated that PASMCs isolated from a well-established rat model of PAH are resistant to apoptosis under both basal and stimulated conditions. The cell membrane protein aquaporin 1 (AQP1) was initially described as a water transport channel, but more recently has been implicated in other cellular functions including migration and proliferation, and in several distinct cancer types, has been associated with apoptosis resistance. AQP1 is upregulated in PASMCs isolated from rat models of PAH suggesting a ‘quasi- malignant’ disease model, although AQP1’s exact role in apoptosis resistance is unclear. Exciting new data from an unpublished proteomics study using lung lysates demonstrates that AQP1 immunoprecipitates with total caspase-3, a enzymatic protein activated in apoptosis which is translocated to the nucleus where it initiates cell death. In proximity studies utilizing biotin ligase techniques, I have demonstrated that AQP1 and total caspase- 3 come within close proximity in live cells. Furthermore, in silico analysis of the AQP1 protein reveals 3 potential caspase-3 cleavage sites, which provide a mechanism for this protein-protein interaction. Together, these data suggest that AQP1 interacts with caspase-3, providing a novel relationship between AQP1, the caspase cascade, and resistance to apoptosis. This application serves to provide a training vehicle as I explore a potential mechanism by which AQP1 regulates apoptosis during PAH. Aim 1 is designed to determine whether the cytosolic caspase-3 cleavage site(s) on AQP1 are necessary for AQP1/caspase-3 interaction, Aim 2 serves to evaluate the impact of AQP1 on nuclear localization of capsase-3, and finally Aim 3 will establish if increased AQP1 is necessary and/or sufficient to confer apoptosis resistance. Techniques utilized to address these aims include but are not limited to protein expression and site directed mutagenesis, biotin ligase proximity assays, co-immunoprecipitation, animal models of PAH and primary cell isolation, immunofluorescence and confocal microscopy, nuclear/cytosolic fractionation, luminescent caspase-3/7 activity assay, Hoechst staining and TUNEL staining. Completion of this project will provide novel insight into the interaction between AQP1 and caspase-3 and the role for AQP1 in apoptosis resistance as well as provide a novel pathway for new therapeutic targets. The skills obtained in the design and execution of this study and the experimental results will provide the necessary foundation for a K award and an excellent platform on which to start a career as an independently funded clinician scientist focused on PAH. Insights gained from this work also could have implications beyond PAH to any diagnosis in which cellular resistance to apoptosis is essential to the development of disease.

NIH Research Projects · FY 2026 · 2022-08

PROJECT SUMMARY This project aims to define the role of farnesylated prelamin A in premature and physiological aging. Genetic mutations leading to defective processing of prelamin A, the precursor for the nuclear scaffold protein lamin A, result in persistence of its farnesyl modification and cause premature aging disorders. The best known of these is Hutchinson-Gilford progeria syndrome (HGPS), in which a truncated farnesylated prelamin A variant called “progerin” is expressed. Mandibuloacral dysplasia-type B (MAD-B) is a related disease, where unprocessed farnesylated prelamin A itself accumulates in cells. Some evidence suggests that prelamin A also accumulates during physiological aging. However, the mechanistic role of farnesylated prelamin A in premature aging disorders remains unclear, and its association with physiological aging has not been rigorously tested. We hypothesize that prelamin A is a driver of osteoporosis and cardiovascular disease that occurs in premature and possibly physiological aging. To specifically examine the consequences of prelamin A accumulation, we generated a novel mouse strain (LmnaL648R/L648R) with a mutation that abolishes its processing by the zinc metalloprotease ZMPSTE24. These mice express solely prelamin A (and no mature lamin A), exhibit profound bone loss, but have a significantly longer lifespan than other progeria models and are thus ideal to study the effects of prelamin A during aging. In Aim 1, we will determine how prelamin A affects the number, function, development, transcriptomes and signaling of bone cells, and compare these to what occurs in physiological aging. We will also assess whether these mice develop vascular disease as they age or develop accelerated atherosclerosis when combined with genetic (Ldlr-/-) and high-fat diet interventions that sensitize mice to atherosclerosis. In Aim 2, we will determine how prelamin A affects cultured cells and define the mechanism(s) by which it promotes cellular alterations related to aging. We will carry out chronological transcriptomic analyses in cultured cells to define the earliest events promoted by prelamin A to distinguish causal changes from chronic responses. We will also test the hypothesis that prelamin A induces nuclear envelope rupture that could stimulate a cytosolic DNA sensor and lead to a transcriptional program of inflammation. We will relate the mechanistic insights obtained from these in vitro experiments to affected cells in the LmnaL648R/L648R mice. In Aim 3, we will determine if prelamin A actually accumulates during physiological aging by examining bone and vascular tissue of young and old mice. We will also probe human tissue arrays and a panel of fibroblasts from young and aged individuals for prelamin A. Elucidating the cellular mechanisms and consequences of prelamin A accumulation could change clinical paradigms for the treatment of prelamin A-based premature aging disorders. More broadly, the results of this project could rigorously implicate prelamin A in physiological aging, identifying it as a potential therapeutic target for age-associated osteoporosis, cardiovascular disease, and possibly other ailments.

NIH Research Projects · FY 2025 · 2022-08

The synovium has recently been described as an important indicator of systemic pathologies and inflammation, suggesting its significance in overall physiological health. It is responsible for regulating joint health and is particularly susceptible to inflammation and age-related changes that lead to disease pathologies such as osteoarthritis (OA). Cellular senescence, or the phenotypic state where cells cease dividing in response to inflammation, injury, or oxidative stress has been implicated as a potential marker and regulator of various systemic pathologies including age-related disease states. While the synovium has also been highlighted as a key sensor for age-related systemic pathology, the role of senescent cells (SnCs) in the synovium is poorly understood. The goal of this project is to use experimental and computational tools, including models of age- and trauma-induced arthritis, to identify the role of senescent cells (SnCs) in the systemic pathology, dysfunction, and tissue degradation of the synovium. We previously demonstrated that SnCs play a central role in the pathogenesis of arthritis, and that their removal may serve as a disease-modifying therapy. However, the identity and function of these cells in vivo, including interactions with the immune system, remain elusive. We do not yet understand the in vivo phenotype of synovial SnCs and the heterogeneity of senescence, which includes both beneficial and pathological cells. Furthermore, the senotypes (subtypes of senescent cells) are likely different across variables such as age, sex, and disease state. In this proposal, we will identify, characterize, and investigate the accumulation of SnCs across multiple variables in the synovium. We will apply computational transfer learning algorithms to understand which cell types become senescent and how these SnCs interact with their environment and regulate disease. We hypothesize that there are multiple types of SnCs that contribute to the systemic pathology, dysfunction, and tissue degradation of the synovium. This research will advance our fundamental understanding of cellular senescence, aging, and arthritis. This work has the potential to address aging-associated factors that may modify therapeutic approaches, providing broad impact and clinical relevance to this work. Our Specific Aims are: Specific Aim 1. Characterize SnCs and senotypes in the synovium in young and aged murine models of arthritis. SubAim 1.1. Perform transfer learning of SenSigs on scRNAseq datasets from young and aged murine models of OA to identify SnC subtypes. SubAim 1.2. Define SnC communication networks with stromal and immune cells. SubAim 1.3. Validate SnC subtypes and cell-cell communication using multiplex fluorescent immunohistochemistry (mfIHC). SubAim 1.4. Identify circulating biomarkers of arthritis senotypes.

NIH Research Projects · FY 2025 · 2022-08

PROJECT SUMMARY Fused in sarcoma (FUS) is a nuclear RNA binding protein that undergoes liquid-liquid phase separation (LLPS). When mislocalized and/or dysregulated, aberrant phase separation of FUS leads to the formation of pathogenic solid-like aggregates that are implicated in neurodegenerative diseases including amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). We have recently discovered that reduction of DDX6, an RNA helicase known to interact with FUS, significantly diminishes cytoplasmic FUS granule formation during stress in neuroblastoma cells (SH-SY5Y). Our preliminary results show that (i) DDX6 modulates FUS condensate number and size in a concentration dependent manner in vitro i.e., DDX6 promotes FUS condensate formation at low concentrations but limits growth above a finite level and (ii) DDX6 forms a discontinuous ring around FUS condensates. Building on these exciting results, I aim to test the hypothesis that DDX6 regulates FUS granules through dual roles as a granule nucleator and Pickering agent. Pickering agents are particles with distinct properties which adsorb to the surface of condensates, promoting their liquidity and maintaining small condensate size. Based on our results, we propose that DDX6 promotes FUS granule nucleation while its role as a Pickering agent maintains small droplet size and liquidity, thus preventing pathogenic aggregation. Additionally, we predict that ATP binding and RNA structure will affect the activity of DDX6 on FUS granules. We will test these predictions in three aims. Aim 1 will utilize in vitro condensation assays to evaluate the interaction between DDX6, FUS, RNA and ATP and to establish whether DDX6 acts as a Pickering agent. Aim 2 will utilize biochemical and single molecule assays to characterize the molecular-level dynamics of the interaction between FUS, DDX6, and RNA, and its dependence on ATP. Finally, aim 3 will use cell-based methods to investigate how tuning intracellular DDX6 concentrations and disrupting ATP binding affects FUS granule formation in wildtype and ALS-associated mutants. Together, this work will lead to a deeper understanding of RNA-protein granule regulation which is of utmost importance to treating neurodegenerative diseases such as ALS and FTD.

NIH Research Projects · FY 2026 · 2022-08

Obesity and its metabolic complications are leading causes of morbidity and mortality in the world. Evidence is mounting that inappropriate timing of food intake contributes to obesity. Late eating is associated with obesity and metabolic syndrome, suggesting that circadian misalignment may be the mechanism underlying the adverse metabolic consequences of late eating. We hypothesize that meal timing in relation to the endogenous circadian rhythm, rather than to clock hour, determines metabolic outcomes. In this study, we will use dim light melatonin onset (DLMO), the gold-standard for ascertaining central circadian output, to assess individual circadian rhythms. We will use DLMO to prospectively assign “early” (DLMO-3h) vs “late” (DLMO+1h) dinner while maintaining the same sleep times (DLMO+2h to +10h) to evaluate whether acute metabolic dysfunction can be reliably induced or prevented by setting dinner times around DLMO. We will use hourly blood sampling for detailed glucose and insulin profiles, oral [2H31] palmitate tracer to quantify dietary fat oxidation, and whole-room indirect calorimetry to measure total fat oxidation. We will enroll both normal-weight healthy adults (NWH) and adults with obesity and prediabetes (OPD), as the latter population is particularly vulnerable to metabolic diseases and could derive immediate benefit from our findings. The specific aims are to: 1) Quantify the impact of DLMO-based “early” vs. “late” dinner time on post-prandial and overnight glucose and insulin levels in NWH and OPD adults, 2) Measure the impact of DLMO-based “early” vs. “late” dinner time on (a) exogenous/dietary and (b) total fat oxidation in NWH and OPD adults, and 3) Examine the utility of circadian phase markers to predict susceptibility to late eating-induced metabolic dysfunction. For Aims 1 and 2, we will crossover-randomize 16 NWH adults (8 men, 8 women) and 16 OPD adults (8 men, 8 women) to the 2 dinner times with isocaloric feeding in a metabolic chamber. For Aim 3, we will leverage validated circadian metrics derived from actigraphy and ingestible thermosensors to predict effects of late dinner. Dr. Daisy Duan’s long-term career goal is to become an independent clinician investigator leveraging novel mechanistic insights that underly the intersection between the circadian system and metabolism to design and validate interventions for the prevention and treatment of obesity and its metabolic complications. She seeks a K23 mentored career development award to gain critical skills and experience in order to effectively lead an independently-funded research program. The goals during the award period include developing expertise in the design and implementation of in vivo metabolic studies and in the principles, practice, and analytical methods in sleep and circadian phenotyping techniques, through a combination of mentored research experience, focused coursework, hands-on learning in research methodology, participation in local and national conferences, grant writing, and leadership training and experience. The proposed study will lay the foundation for novel, circadian- based meal timing as a precision medicine approach to obesity and metabolic dysfunction.

NIH Research Projects · FY 2025 · 2022-08

Abstract/Summary The development of methods for molecule construction and functionalization has expedited the drug development process during the past decades. To this end, enzymatic C–H functionalization represents a powerful approach to enable rapid molecular construction. Harnessing the ubiquitous C–H bonds in organic molecules and genetic tunability of protein catalysts, enzymatic C–H functionalization provides sustainable strategies to effect chemical transformations with high selectivity and efficiency. Despite significant advances, the reaction scope of current enzymatic C–H functionalization is restricted to chemical reactivities acquired through natural evolution. This limitation hampers the applications of enzymes for drug development as many medicinally important chemical motifs are rarely present or even completely absent in biology. To expand the chemical space of biosynthesis, we herein aim to develop new biocatalytic systems by repurposing metalloenzymes for C–H functionalization reactions currently not present in biology. By integrating protein engineering, computational modeling, organic synthesis, and biochemical and inorganic spectroscopic analysis, we will create collections of metalloenzyme catalysts that can directly functionalize inert C(sp3)–H bonds to install biomedically relevant chemical moieties. We envision that the enzymes we create will not only provide powerful genetically encoded tools for numerous synthetic and biological applications, but will also offer a fertile ground to enrich our fundamental knowledge of biochemistry and enzymatic catalysis

NIH Research Projects · FY 2025 · 2022-08

Huntington’s disease (HD) is a dominantly inherited, fatal neurodegenerative disorder caused by a CAG expansion in the Huntingtin (HTT) gene. HD preferentially involves the basal ganglia- especially the striatum- but also affects other brain regions and has no cure or disease-modifying treatment yet. Because of its gain-of- function mechanism, strategies to lower mutant HTT are promising as first-ever disease-modifying therapies. Most approaches are currently targeted at manifest HD when clinical outcomes can be used to evaluate the effectiveness. However, as almost 50% of striatal volume has been lost at the time of onset, it would be preferable to begin treatment in the premanifest period before massive loss of striatal volumes. An unmet challenge is how to reliably evaluate therapeutic efficacy in the absence of clinical symptoms as outcome measures. The clinical diagnosis of HD is based on the presence of movement disorders. However, functional changes in the brain can precede motor onset by many years. Neurovascular abnormalities have been reported in premanifest and early HD by us and others. We have reported significantly altered arteriolar CBV (CBVa) in premanifest HD brains, when striatal atrophy was undetectable. We recently found that altered CBVa occurred prior to striatal atrophy in an HD mouse model and that CRISPR/Cas9-mediated mHTT lowering restored CBVa in premanifest HD mice. Collectively, these data suggest reliable measures of neurovascular changes might be valuable biomarkers in premanifest HD. CBV is strongly coupled with brain metabolism, and cerebral metabolic abnormalities are increasingly considered as early neuropathological events in HD. We found impaired response of cerebral metabolism to visual stimulation in premanifest HD patients, correlating with the CAG-Age product (CAP) score, supporting that metabolic disturbances occur at early pathogenic stage and may be another early biomarker for HD. In addition, the recent (re)discovery of brain lymphatic vessels and CSF-lymphatic drainage system illustrates an important brain waste clearance system. Our preliminary data implicate that impairment of elements of this system precedes striatal atrophy and mHTT aggregation in HD mice. The objective of this project is to identify early brain functional changes that rapidly respond to treatment in HD preclinical study by incorporating multimodal advanced MRI measures and to develop sensitive biomarkers translatable to HD clinical trials, particularly in the premanifest period. Aim 1. We will test the hypothesis that mutant HTT impairs cerebral metabolism and alters neurovascular responsivity prior to brain atrophy and behavioral deficits in HD mice. Aim 2. We will assess glymphatic function by measuring lymphatic flow dynamics and BBB permeability in HD mice. Aim 3. We will determine the extent to which metabolic, vascular, and lymphatic MRI measures can monitor the effects of premanifest and manifest treatment of HD mice with CRISPR/Cas9-mediated HTT lowering.

NIH Research Projects · FY 2025 · 2022-08

SUMMARY The staggering incidence of opioid addiction continues at epidemic levels, disrupting and destroying the lives of millions of Americans with a yearly financial toll nearing $80 billion. A major hurdle in treating opioid addiction is a chronic cycle of withdrawal and relapse. Rodent studies have identified long-term changes in gene expression, epigenetics, and circuit connectivity after opioid exposure and withdrawal, but few studies have investigated the interaction of these changes across modalities. We recently developed a new class of brain mapping tools based on cellular barcoding that can relate the three modalities in single cells and experiments. These tools, which include MAPseq and BARseq, use RNA barcodes to uniquely label thousands of neurons per experiment and map their inter-regional connections. In each labeled neuron, barcodes are trafficked into the axons, where we can detect them by high-throughput sequencing. Matching up barcode sequences across potential target regions then produces the single-cell projection matrix for all barcoded neurons. As barcodes are mRNAs, they are in the same modality as the endogenous transcriptome, allowing us to natively bridge single-cell connectomic measurements with single-cell measures of gene expression and genome accessibility in the same cells. Finally, combination of these technologies with spatial transcriptomics methods, including STARmap and BARseq2, lets us map the observed changes with high resolution in 3D brain space. Here we apply our multi-omic tools to measure the changes induced in the medial orbitofrontal cortex (mOFC) in two rodent models of opioid withdrawal and relapse. Our collaborators in the Shaham lab recently identified opposing functional connectivity changes in rat mOFC after electric barrier-induced voluntary abstinence. In the same rat model using our tools, we will define which cell types are responsible for these connectional changes, how single-cell connection patterns change, and how gene expression, epigenetics, and connectivity changes interact at the level of single cells. Critically, we define cell types holistically across transcriptomic, epigenomic, and connectomic modalities. We then compare these changes to those observed in mOFC in a second rodent model, a classical opioid withdrawal conditioned place aversion (CPA) model in mice. We hope to identify a set of changes robust to the two behaviors and conserved across species, highlighting them as core features of opioid withdrawal and relapse. We will causally manipulate epigenetic factors associated with these core changes by cell-type-specific knockouts and overexpression, and assess their effects on behavior. The public health impact of this work lies in the fundamental understanding of the long-term circuit changes induced by opioid withdrawal and relapse at unprecedented multi-omic resolution and the potential identification of a set of conserved core changes as candidates for treatment.

NIH Research Projects · FY 2025 · 2022-07

The overarching goal of the proposed aims has been to leverage novel methods with large and underutilized data sets to evaluate the potential impact of increasingly specific HIV responses across generalized epidemic settings in Sub-Saharan Africa (SSA) in reducing overall HIV incidence. This application was highly responsive to multiple areas of interest in the recent Notice of Special Interest (NOSI): Harnessing Big Data to Halt HIV (NOT-AI-21- 054). Moreover, the aims aligned with current realities of the HIV pandemic. While overall incidence has steadily declined over the last 15 years, over 1.5 million people newly acquired HIV in 2020 including one million people across SSA. The risk for HIV is not evenly distributed anywhere in the world. And while specific key populations are recognized to be at increased risk of HIV in many higher income settings, a general population construct is often used to represent HIV epidemics across SSA. This construct typically negates proximal determinants of HIV acquisition and transmission and ultimately has limited the effectiveness of the HIV response domestically in the US and around the world. We proposed an ambitious set of aims to leverage available HIV-related data for key populations as well as auxiliary data including from social media, search patterns, spatial data, socioeconomic and migration data. We are assembling multiple data sources and integrating these data to build a comprehensive data warehouse to estimate key population-specific indicators including HIV incidence and prevalence, population size, engagement in the HIV treatment cascade, and structural determinants. These estimates, augmented by small area estimation methods where data are sparse, will inform dynamic transmission models to estimate differential risks of onward HIV transmission among key populations and to better address the needs of key populations compared with general-population approaches. Finally, we are leveraging very large and underutilized existing program data for HIV testing, prevention, and treatment programs. This was done in partnership with implementing partners. Cameroon, Kenya, Senegal, and South Africa are used as exemplar countries with sufficient data, willing governments, and representation of common HIV epidemic typologies in their respective regions of SSA. The following aims are near completion and will proceed without any foreign subawards.

NIH Research Projects · FY 2026 · 2022-07

In the United States, heat is responsible for more fatalities than any other type of weather and the burden of morbidity and mortality attributable to heat is growing. Southern cities, such as New Orleans, are expected to suffer some of the greatest consequences of extreme heat in the coming decades. Communities need help measuring and predicting the impacts of heat to dedicate appropriate resources to protect public health and ensure public safety. This challenge is particularly acute in New Orleans, where little is known about the relationship between heat exposure and health outcomes (i.e., the heat metric or threshold that is most predictive of morbidity and mortality). Further, because this relationship can be modified by local meteorological patterns, housing characteristics, individual behaviors, and other neighborhood-level factors, location-specific data and analysis is needed to inform public health and safety investments. In the proposed project, we will (1) determine which exposure metrics, thresholds, and duration predict heat-related morbidity and mortality within New Orleans and use this to estimate the current public health burden of heat exposure to children, adults, and the elderly; (2) identify individual and neighborhood characteristics that increase vulnerability to heat-related morbidity and mortality and create an empirically-based heat-health vulnerability index for New Orleans; (3) collect and analyze citizen-sourced measurements of indoor and outdoor thermal environments, social media observations, and “real time” assessments of heat, risk perceptions, and health symptoms through ecological momentary assessment to further characterize small-area variation in heat exposure and human behaviors that may lead to increased heat vulnerability; and (4) in collaboration with a community advisory panel, develop city-wide recommendations for improved heat-health preparedness in New Orleans and translate our findings to other communities with the production of a toolkit on heat-health preparedness. Key innovations of this project include the use of novel data sources and methods, such as fine- scale gridded data and citizen-sourced measurements of temperature and humidity, to more accurately characterize personal heat exposure; a comprehensive assessment of the burden of heat across the life course to include symptoms of heat stress, sleep quality, hospitalizations, and mortality ascertained through a combination of healthcare claims analysis and ecological momentary assessment; and a dedication to community engagement and research translation throughout the life of the project to ensure that research results lead to policy action and public health and safety improvements.

NIH Research Projects · FY 2025 · 2022-07

ABSTRACT Jing Sun is a physician epidemiologist and Assistant Scientist in the Department of Epidemiology at Johns Hopkins Bloomberg School of Public Health. She seeks a mentored career development award to fill knowledge gaps in her current training, allowing her to develop as an independent molecular epidemiologist who can bridge the diverse fields of HIV epidemiology, genetics / epigenetics, and aging to improve long-term outcomes among people with HIV (PWH). Since the advent of effective antiretroviral therapy, there has been a rising burden of aging-related conditions among PWH. Mitochondrial damage is a major driver of aging, but its role in HIV and aging has been understudied. Leveraging data, infrastructure, and the biospecimens of two long-term HIV cohorts (ALIVE and MACS), Dr. Sun will define the relationship between mitochondrial DNA copy number, a noninvasive marker of mitochondrial content, and biological aging at the phenotypic, molecular, and immunological levels among people with and without HIV (Aim 1). She will further define the interrelationship of mitochondrial DNA copy number with DNA methylation and nuclear genetics using existing data from ALIVE and MACS (Aim 2). In Aim 3, she will design a study nested within an ongoing exercise intervention trial to evaluate the effect of exercise on mitochondrial function, methylation patterns, and physical function among PWH. To complete these research aims and develop an independent research program, Dr. Sun will receive training in the following areas: (1) statistical genetics and multi-omics analysis; (2) laboratory measurement of mitochondrial function; (3) biobanking and management; (4) research leadership. The training program consists of didactic coursework and seminars, hands-on practice in statistical genetics analysis, informational laboratory visits and shadowing, participation in a certificate program, and mentored research by an established and diverse team of senior investigators. Completion of the proposed aims will directly address important epidemiological, mechanistic, and interventional knowledge gaps and elucidate the role of mitochondrial function on biological aging among PWH. Findings will provide evidence for targets of future clinical and interventional research with the potential to guide long-term management and improve quality of life among PWH. Preliminary data and research capacity built through this K01 award will support an R01 investigating the role of mitochondrial function, aging, and comorbidities among PWH using a multi-omics approach. It will further support Dr. Sun’s long-term career goals to become an international leader in HIV and aging with content expertise in mitochondrial genetics and function.

NIH Research Projects · FY 2025 · 2022-07

Subpopulations within the US have disproportionate hypertension (high blood pressure; HBP) rates – differences that have persisted for decades and at the highest cost to society of all cardiovascular conditions. The underlying cause of these differences is unknown, and previous studies have mostly focused on individual-level behaviors, stressors, and physiologic risk factors leaving a missed opportunity to uncover and address the underlying causes of these disproportionate rates. Factors at both the individual-level and area-level may play a role; thus, to fully address hypertension in our nation, we must investigate the pathways through which both individual-level and area-level factors influence HBP risk factors and rates. Given that many key decisions about health and healthcare services are made at the level of the county-seat, county-level factors are critical to assess. Using a novel 5-domain measure of county-level social conditions, our previous cross-sectional studies have demonstrated that our index is associated with higher BMI, one behavioral risk factor for HBP; however, this work has left gaps in understanding how county-level factors influence other risk factors for HBP and HBP rates. Our goal is to conduct a multi-level national study to investigate associations between our novel multi-dimensional measure of county-level social and economic factors and: physiologic, behavioral, and structural risk factors for HBP (Aim 1), HBP incidence, prevalence, and severity (Aim 2), and how much counties could save in HBP healthcare costs if county-level social and economic conditions were modified (Aim 3). We leverage pre-existing resources that are uniquely available to us: (a) our published county-level index, (b) US News & World Report hospital rankings of healthcare quality, and (c) longitudinal behavioral and biomarker HBP data from 30,239 adults across the US in the REGARDS cohort study. We expand beyond previous studies by using a multi-domain measure of county-level factors, across urban and rural areas, applying it to longitudinal health data that allows us to assess exposure to these factors at multiple times in the lifecourse, and quantifying how much social and economic conditions cost counties in HBP healthcare spending if they go unmitigated. We will translate our findings into policy briefs targeted toward county-level executives in the US. Our team of experts in cardiovascular disease (CVD), social and clinical epidemiology, and health economics, with representation from REGARDS, two Hopkins Centers focused on vulnerable populations, Hopkins’ CVD Epidemiology, and former county leaders, are well-equipped to execute this New Investigator application. Our results will offer evidence of how county-level factors influence HBP, with the potential to build support for county-level policy decisions that can ultimately reduce HBP rates.

NIH Research Projects · FY 2025 · 2022-07

Abstract Survival studies show marked racial disparity effect in head and neck cancer between African Americans (AA) and whites. AA may present with more advanced disease and have twice the age-adjusted mortality rate compared with whites. Immune checkpoint inhibitors offer new hope for some patients with recurrent or metastatic disease, but further improvement in therapy is contingent upon developing a comprehensive immunogenetic map of neoplastic evolution in HNSCC. Nearly 20% of patients with oral cancers harbor multiple pre-malignant lesions showing signs of dysplasia, often visually identified as leukoplakia. As some of these lesions evolve to malignant neoplasms, they represent intermediate steps in HPV negative oral squamous cell carcinoma (OSCC) progression. Genetic changes arising at the earliest stages of tumor development drive tumor progression and curtail the propensity of the immune system to destroy precancerous cells. Genetic aberrations selected during OSCC evolution can create a dysfunctional tumor immune microenvironment by upregulating key immunomodulatory ligands that induce immune tolerance and T cell exhaustion. The role of cross-talk between neoplastic cells and their immune microenvironment, particularly in its early developmental stages, has yet to be elucidated. Moreover, little baseline information exists in these lesions in AA, and even less is known about the key genetic changes that lead to progression and immune invasion in this population. The central premise of this project is that key racial differences in both inherited and somatic genetic changes during the progression of OSCC impact the expression of key immunomodulatory cytokines or ligands within the tumor microenvironment in order to escape an antitumor immune response. The specific aims of this project will use whole-exome sequencing, RNA sequencing, high-throughput computational analyses, and tissue cell localization methods to map the specific mutational patterns and corresponding immune landscape represented by expression of key immunomodulatory ligands and signatures of immune tolerance and T cell exhaustion in lesions along the pathway of oral tumorigenesis. This data will help us understand the biologic underpinnings of the different progression pathways and interactions with the immune microenvironment. Such mapping should provide crucial insights that have significant implications for risk assessment, tumor surveillance and treatment interventions for OSCC in AA patients.

NIH Research Projects · FY 2025 · 2022-07

There has been critical underinvestment in addressing the Substance Use Disorders (SUDs) problem. We continue to rely on the same ineffective approaches. In this proposal, we promote and support disruptive innovations to stem the rising tide of SUDs. Such disruptions arise from clinician-scientists who are treating and researching SUDs yet are unaware of, or do not see value in, commercializing their ideas. Our overarching goals are to encourage more scientists to think entrepreneurially and to provide access to the resources they need for commercializing their ideas. Through our proposed tasks, we will develop and deliver entrepreneurship education specifically tailored for basic scientists to empower them with the tools for their commercialization journeys to advance the prevention, diagnosis, and treatment of SUDs (Aim 1). The Innovation for Substance Use Disorders (I4SUD) curriculum will be scientifically developed, implemented, and evaluated during this 5-year grant. The cornerstone of the proposed initiative is a new Certificate in Entrepreneurship at Johns Hopkins University Carey Business School (Aim 2). This program leverages our deep experience teaching entrepreneurship to scientists at Johns Hopkins Medicine to customize content for SUD researchers. The asynchronous online modules developed for the certificate program will be widely distributed (Aim 1). Through our educational efforts, we will intentionally build a community that can support the translation of discovery to market to change the trajectory of the SUD crisis in our nation (Aim 3). This is a specialized program for SUD researchers at all career levels, grounded in theoretical frameworks and complemented by professional education experiences related to entrepreneurial skills formation and community building. I4SUD will consist of three parts: asynchronous online learning modules covering topics in ideation, market discovery, intellectual property, regulatory strategy, revenue models, and business and financial modeling; an in-person multi-day workshop comprising observations in SUD clinics and recovery centers, conversations with stakeholders, designing and prototyping solutions; and mentoring by subject matter experts, culminating in a pitch competition. Program graduates will be equipped to apply for translational funding, patents, IP licensing, or startups. They will belong to a large virtual community of researchers and practitioners while contributing to healthcare entrepreneurship in their academic institutions. With a clear understanding of the gaps that exist in the SUD field, the possibilities for innovation, and the training process needed to help scientists move their discoveries to market, this program will significantly impact public health by facilitating the launch of projects that offer the next generation of solutions to SUD.

NIH Research Projects · FY 2026 · 2022-07

Project Summary: White matter hyperintensities (WMHs) of the brain are a phenotype of cerebrovascular small vessel disease (cSVD) and estimated to represent 40% of the cSVD disease burden . The WMH phenotype is closely associated with damage to the neurovascular unit and is thought to contribute to cognitive impairment, deficits in motor control, and stroke. Understanding how WMHs are affected by vascular risk factors and progress over time is a part of a national research priority of understanding vascular contributions that influence cognitive decline and dementia (VCID). There is a knowledge gap about how WMHs progress in younger age groups (age 40-50). This study addresses two issues of significance regarding early progression: (i) how periventricular WMHs (PVWMHs) and subcortically located deep WMHs (DWMHs) progress independently and influence different clinical outcomes and (ii) how increasing blood–brain barrier (BBB) permeability and microvascular changes are associated with lesion expansion. This study adds to the understanding of the pathophysiology of WMH progression, supporting the development of earlier prevention and mitigation. GeneSTAR is a vascular study of asymptomatic younger (age 40-50) siblings of patients with premature coronary artery disease enriched for WMH. The GeneSTAR population is deeply phenotyped for vascular properties, has a high prevalence of asymptomatic WMHs, and is relatively young at the time of first MRI. We will leverage these features and our experience studying PVWMH, DWMH, and BBB permeability by recalling these participants and repeating the 3T MRI volumetrics with adding two independent measures of BBB permeability. This study will include relatively young African American and European American adults (n=500) with a mean age of 50.8±10.4 at baseline in GeneSTAR and will now reimage them at age 61.9±10.5 years. We will relate any changes in WMH volume to: (i) vascular risk factors (hypertension, diabetes, blood lipids, smoking), (ii) changes in vascular function (carotid artery wall stiffness, pulse pressure, and brachial arterial diameter), and (iii) changes in cognitive function (immediate memory, working memory, information retrieval, multi-domain cognitive skills, and motor function). We also will recall 130 of our proposed participants in years 4 and 5 to study the relationship between WMH progression and increases in BBB permeability and mean transit time (MTT) using serial imaging. This second MRI at year 3 or 4 has the goal of measuring the interval change in all MRI phenotypes measured at year 1 and relate WMH volume progression to BBB permeability and mean transit time measured in year 1 to that at years 4 or 5. We will study how changes in BBB permeability and perfusion are: (i) related to WMH volume changes, (ii) affected by vascular risk factors, and (iii) associated with cognitive changes.

NIH Research Projects · FY 2025 · 2022-07

PROJECT SUMMARY Amyotrophic lateral sclerosis (ALS), an adult onset motor neuron degenerative disease, is increasingly recognized to have clinical, pathological and genetic overlaps with Frontotemporal dementia (FTD). Dysfunction of RNA metabolisms has emerged to play crucial roles in disease etiology. Pathological inclusions and/or causative genetic mutations in several RNA-binding proteins are widely found in the two diseases, such as TDP- 43 and FUS/TLS. Alternatively, the hexanucleotide repeat expansion in the non-coding region of C9ORF72 gene could also induce toxicity from the repeat RNA-derived products. This supports the susceptibility of neurons to the dysfunction of RNA processing and the importance of RNA homeostasis in preserving neuronal integrity. RNA modifications have recently emerged to play important roles in posttranscriptional regulation of gene expression. N6-methyladenosine (m6A) is by far the most abundant internal RNA modification of eukaryotic cells. m6A modification is dynamic and reversible, providing an additional layer of regulation on RNA. It is noted that m6A is most enriched and the methylome is highly specific in the nervous system compared to other tissues. Emerging studies indicate the important roles of m6A in regulating brain function, from development to synaptic plasticity, learning and memory, and neurodegeneration. But it has not been explored in ALS/FTD or other neurodegenerative diseases. The hexanucleotide GGGGCC repeat expansion located in the first intron of the C9ORF72 gene is the most common cause of both ALS and FTD. The leading hypothesis for the disease mechanism is gain of toxicity from the expanded repeats, with two non-mutually exclusive mechanisms: 1) RNA foci formed by repeats that could sequester RNA binding proteins and disrupt RNA processing; and 2) accumulation of dipeptide repeat proteins (DPRs) produced by repeat-associated non-AUG translation (RAN translation). Our lab recently identified that m6A RNA methylation can affect the DPR levels produced from the repeats. Furthermore, we also found that the m6A pathway is profoundly reduced in C9ORF72-ALS/FTD patient neurons. Therefore, it is important to determine the molecular mechanisms on how the m6A dysregulation changes the repeat RNA metabolisms and disturbs the global mRNA processing, and how this contributes to the neuronal dysfunction and degeneration in C9ORF72-ALS/FTD. Furthermore, we will also determine whether targeting specific components in the m6A pathway can rescue the disease-related phenotypes. This study addresses an emerging theme of RNA regulation that has not been explored in neurodegeneration. The findings will help understanding the etiology of the disease and developing novel therapeutic strategies for ALS and FTD.

NIH Research Projects · FY 2025 · 2022-07

Project Summary Brain tumors are relatively common in children, with medulloblastoma (MB) the most frequent type, especially in children under five. MBs can spread through the cerebrospinal fluid, and metastases are common at the time of diagnosis. Some of the regulatory genes, signaling pathways, and gene regulatory networks in MB are known, but the role of non-coding RNAs in MB, particularly long non-coding RNAs (lncRNAs), are poorly described. By applying machine learning to publicly available RNA-seq datasets, we found that lnc-HLX2-7 was highly upregulated and specific for difficult-to-treat and poor prognosis grade 3 (G3) MBs compared with other molecular subgroups. CRISPR-Cas9 depletion of lnc-HLX-2-7 in G3 MB cells significantly reduced cell proliferation, invasion, and 3D colony formation and induced apoptosis. When lnc- HLX-2-7-deleted G3 MB cells were injected into the mouse cerebellum, they produced considerably smaller tumors than those derived from parental cells. Further, cerium oxide nanoparticle-coated antisense oligonucleotides (ASOs) against lnc-HLX2-7 reduced in vivo tumor growth. Our preliminary results also demonstrate that lnc-HLX-2-7 is a critical MB metabolic regulator that modulates NAD+ (nicotinamide adenine dinucleotide) via nicotinamide phosphoribosyltransferase (NAMPT), a key enzyme mediating NAD+ production. The above results highlight the functional impact of lnc-HLX-2-7 on G3 MB development, some of the underlying mechanisms of action, and its importance as a therapeutic target. Our central hypothesis is that lnc-HLX-2-7 is an important oncogenic molecule that can be therapeutically targeted in G3 MBs. We propose the following three Specific Aims to test our hypothesis: (a) to test pre-clinical therapeutics targeting G3 MBs; (b) to delineate the lnc-HLX-2-7-driven molecular mechanisms underlying G3 MB tumors, and (c) to identify how lnc-HLX-2-7 is regulated and regulates other genes in G3 MBs. This study will provide valuable mechanistic insights into how lnc-HLX-2-7 drives G3 MB development, advance pre-clinical therapeutic targeting of this challenging subgroup, and anticipate compensatory and resistance mechanisms. This study provides important insights into how lncRNAs function as critical oncogenes in the brain and other cancers.

NIH Research Projects · FY 2025 · 2022-07

Project Summary Nucleotide repeat elements, including microsatellites or short tandem repeats, are common in eukaryotic genomes. Expansions of short nucleotide repeats have been linked to over 50 different types of genetic disorders, primarily neurological and neuromuscular disorders. Our understanding of how these repeat elements in the human genome cause diseases is still in its infancy. A hexanucleotide repeat expansion in a noncoding region of the C9orf72 gene has been linked to the neurodegenerative diseases amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). ALS is characterized by loss of motor neurons, and the C9orf72 hexanucleotide repeat expansion represents the most common genetic cause of both familial and sporadic ALS. FTD is characterized by degeneration of the frontal and temporal lobes of the brain and is the second-most common type of dementia in people older than 65; the C9orf72 hexanucleotide repeat expansion is also the most common genetic cause of FTD. This repeat expansion is also found to contribute to Alzheimer’s disease and Huntington’s disease. To help relieve the public health burden associated with these diseases, it is important to understand the mechanisms underlying their pathogenesis. Multiple hypotheses exist to explain the pathogenic mechanisms underlying C9orf72-linked ALS/FTD. The goal of the proposed project is to elucidate novel mechanisms through which the C9orf72 hexanucleotide repeat expansion leads to molecular defects and neuronal toxicity, focusing on gain-of-function mechanisms. The specific aims are to identify previously unknown pathogenic cascades initiated by the C9orf72 hexanucleotide repeat expansion. These novel pathogenic cascades include, but are not limited to, RNA toxicity and non-canonical translation products resulting from the C9orf72 hexanucleotide repeat expansion. We propose a series of fundamental studies that combine biochemical, molecular, and genetic approaches to shed light on the novel pathways leading to ALS/FTD pathogenesis and to identify potential intervention strategies. Successful completion of the project is expected to provide insights into fundamental mechanisms of neurodegeneration in ALS/FTD that may ultimately lead to novel approaches for treating ALS/FTD and other relevant neurodegenerative diseases.

NIH Research Projects · FY 2025 · 2022-07

Most disease associated GWAS variants have relatively modest effects on expression in reporter or CRISPR perturbation assays. In addition, enhancer disruption in vivo often has surprisingly weak phenotypic consequences. We hypothesize that a critical missing element is our lack of quantitative models of how multiple TFs interact at an enhancer, and how multiple enhancers interact at a locus to respond to perturbations in a nonlinear way through altered gene network activity. Predicting the impact of genomic variation thus requires quantitative modeling of how one variant's impact depends on other variants through their combined effect on altered cellular regulatory state. The central aim of this proposal is to develop computational methods to infer quantitative models of these combinatorial interactions by training on temporally-resolved measurements of gene activity, enhancer activity, and core cell fate-regulating transcription factor (TF) activity across cell state transitions in early human development. Our preliminary studies show that while promoter knockdown has robust effects on target gene expression, individual enhancer knockdown is often weaker and affects temporal transition dynamics, but not the final steady state. We show that gene network models based on sequence-based machine learning are consistent with these observations. We propose improvements to our sequence based models to develop kinetic rate equation and stochastic simulation gene network models to predict the variable and often temporal effects of enhancer perturbation. We will generate high time resolution ATAC, H3K27ac, and scRNA-seq data to train these models, and validate the gene network predictions of network response with CRISPRi in a native genomic context. We will first focus on our embryonic- stem-cell to definitive-endoderm (ESC-DE) system, and we will then develop methods to generalize application of these focused models to larger ENCODE regulatory datasets. Our work will enable a quantitative understanding of how the altered activity of regulatory elements affects the stability and dynamics of the gene regulatory networks within which the element operates, and how they play a role in controlling developmentally important and disease relevant cell state transitions.

NIH Research Projects · FY 2025 · 2022-07

Abstract. In many low- and middle-income countries (LMICs) breast cancer is diagnosed at advanced stages. Many women present with a palpable breast mass, which is rare in communities where breast screening is available. In addition to limited imaging facilities, prolonged diagnostic delays (76-630 days) due in part to the extreme shortage of pathologists (as few as 1 per million/population) contribute to a 5-year mortality rate up to 4 times higher than that in the US. An innovative solution to this problem could be an affordable, easily deployable molecular test to identify and prioritize women likely to have a malignancy for expedited biopsy and pathology review. It is well established that early detection of breast cancer improves survival. With our industrial partner, Cepheid, we propose to build on our published breast cancer detection prototype to develop an affordable, <3- hour, automated breast cancer detection (aBCD) assay that analyzes a panel of hypermethylated genes in breast fine needle aspirates (FNAs).The proposed innovations will cut the assay time in half, and reduce costs by at least 3-fold to provide a single-cartridge assay for quick cancer detection. In Aim 1a, we will optimize the Offboard bisulfite-mediated DNA conversion method and test its efficiency in Patient Set 1 FNAs (N= 29 malignant, 25 benign). In Aim 1b we will select one optimal 5-marker panel and test its performance using first, the gold standard, FFPE samples (N= 30 malignant, 30 benign), and then, Patient Set 2 FNAs (N=35 malignant, 35 benign). In Aim 1c, we will perform technical validation of the aBCD assay. Intra-assay reproducibility will be assessed on multiple sample collections of Patient Set 3 FNAs (N=30 malignant, 30 benign). Inter-operator reproducibility will be determined using replicate FNA slides from Patient Set 2 (N= 35 malignant, 35 benign). The goal of Aim 2a is to perform clinical validation of the aBCD assay. We will first select a threshold in a Training set of FNAs: Patient Set 4 (N=100 malignant, 100 benign) to optimally balance sensitivity and specificity, and validate performance of the selected threshold in a Test set of FNAs: Patient Set 5 (N= 180 malignant, 180 benign). We will measure the accuracy (sensitivity, specificity, and positive- and negative-predictive value) of aBCD-based diagnosis to distinguish benign versus malignant lesions using histopathological diagnosis of the core biopsy as the gold standard. Lastly, in Aim 2b, to determine whether select patient characteristics alter the performance of the aBCD assay, we will test its clinical accuracy among specific patient subgroups based on age, race, BMI, and tumor characteristics (grade, stage, tumor subtype). All these steps are necessary to ensure an accurate and reliable test. This intervention is paradigm shifting, and could revolutionize the current detection of breast cancer in underserved regions of the world by expedited treatment and, in turn, saving thousands of lives yearly. This study will also facilitate further development of the aBCD assay toward commercialization, making it accessible globally.

NIH Research Projects · FY 2025 · 2022-07

The 2021-2025 HIV National Strategic Plan (HIV Plan) recognizes stigma as a barrier to the success of the Ending the HIV Epidemic (EHE) initiative given consistent data highlighting the relationship between stigma and suboptimal HIV testing, prevention, and treatment outcomes. The prioritization of stigma by government agencies has yet to translate to scaled stigma mitigation strategies as a means of improving mental health, quality of life, and optimizing the HIV response. In response, our goal is to address the disconnect between practice and priorities in HIV epidemic control by systematically collecting stigma data in partnership with seven health departments and community partners in priority EHE areas, systematically visualizing key stigma indicators and the potential impact of stigma mitigation on EHE pillars and local HIV incidence, and using implementation research to optimize the usability of these tools and inform stigma mitigation interventions. These goals will be achieved via the following specific aims: Specific Aim 1 – Use historical and future iterations of data from AMIS, TWIST, PLHIV Stigma Index 2.0, NHBS, combined with systematic reviews to create stigma dashboards integrated into AIDSVu. Specific Aim 2 Using stigma data to model and visualize the effects of stigma and stigma mitigation interventions on proximal EHE indicators including incidence, knowledge of status, diagnoses, linkage to HIV care, viral suppression, and PrEP coverage. Specific Aim 3 – Use qualitative methods with health departments and CBO partners to inform the optimization of the Stigma Dashboard including usability, presentation, and information included to inform stigma-related policies and the development of local stigma mitigation strategies. These aims are highly responsive to “Respond: Epidemiology to End the HIV Epidemic” in using data to support effective and tailored approaches to respond to local epidemics and include balanced collaborations between epidemiologists, data scientists, statisticians, and local health departments and community partners. The key outputs of the proposed research will include refined, user-centered stigma dashboards integrated into AIDSVu that allow for data visualization and tools to plan and assess downstream impacts of intervention, as well as guidelines to support implementation of use of stigma dashboards and support connection to evidence for stigma mitigation intervention planning to achieve HIV epidemic control goals in additional health departments. While an ambitious research agenda, the diverse experience and competencies of this investigative team focused on stigma and existing partnerships makes us well placed to be successful in contributing to the Ending HIV Epidemic plan through improved measurement, use of, and response to HIV-related and intersectional stigmas.

NIH Research Projects · FY 2025 · 2022-07

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 Biochemistry, Cellular and Molecular Biology (BCMB) Graduate Program has trained more than 700 scientists since 1972. BCMB serves as the major training program for seven basic science departments at the Johns Hopkins University School of Medicine and is one of the oldest multidisciplinary graduate programs in the country. There are 100 faculty members actively involved in research, teaching and as mentors. The Departments that participate in the program are Biological Chemistry, Biophysics and Biophysical Chemistry, Cell Biology, Molecular Biology and Genetics, Neuroscience, Pharmacology and Molecular Sciences, and Physiology. An average of 22 students matriculates each year and obtain their Ph.D. in an average of 5.7 years. The objectives of the BCMB program are: (1) to provide a broad and deep science curriculum; (2) to provide longitudinal training in rigorous, reproducible and responsible experimental research; (3) to provide training in professional skills; (4) to provide activities for trainees to explore career options; and (5) to recruit and support students into a rigorous training environment that is positive, supportive, and safe. These objectives will be met through a rigorous curriculum covering the first year of study that includes courses focused on macromolecules (energetics, structure and function), molecular biology and genomics, genetics, organic mechanisms in biology, cell structure and dynamics, and pathways and regulation. There is a new strong emphasis on bioinformatics and computational skills throughout the coursework. There are multiple paper discussion courses extending from years 1 through 3 to support training in rigorous and reproducible research. There are discussion courses that focus on responsible conduct of research, in addition to department-based workshops as students advance in their thesis work. Oral and written presentation skills are developed throughout training through workshops targeting different specific tasks (oral exams, chalk talks, manuscript and proposal writing). Professional development and career planning is an integral part of the program, occurring through workshops and courses throughout the training period, as well as diverse internship offerings. Most students publish multiple research papers and the training concludes with presentation of a public seminar and submission of the doctoral thesis. BCMB graduates hold leadership positions at all levels of academia, government and industry. The success of our students is fostered by an extraordinary level of collaboration and interaction among the faculty and trainees. Special emphasis is placed on applying conceptual breakthroughs in basic science to problems relevant to human health and disease. Here we request 25 training grant slots to appoint training grant eligible students during their first two years in the program.