Translational Biomedical Research Research Articles

The Chemical Probes Portal: an expert review-based public resource to empower chemical probe assessment, selection and use.

We describe the Chemical Probes Portal (https://www.chemicalprobes.org/), an expert review-based public resource to empower chemical probe assessment, selection and use. Chemical probes are high-quality small-molecule reagents, often inhibitors, that are important for exploring protein function and biological mechanisms, and for validating targets for drug discovery. The publication, dissemination and use of chemical probes provide an important means to accelerate the functional annotation of proteins, the study of proteins in cell biology, physiology, and disease pathology, and to inform and enable subsequent pioneering drug discovery and development efforts. However, the widespread use of small-molecule compounds that are claimed as chemical probes but are lacking sufficient quality, especially being inadequately selective for the desired target or even broadly promiscuous in behaviour, has resulted in many erroneous conclusions in the biomedical literature. The Chemical Probes Portal was established as a public resource to aid the selection and best-practice use of chemical probes in basic and translational biomedical research. We describe the background, principles and content of the Portal and its technical development, as well as examples of its applications and use. The Chemical Probes Portal is a community resource and we therefore describe how researchers can be involved in its content and development.

Cancer prediction using RNA sequencing and deep learning

Numerous areas of medical services, including as imaging diagnostics, advanced pathology, emergency clinic confirmation prediction, drug plan, grouping of cancer and stromal cells, specialist help, and so forth, have profited from the utilization of deep learning. Cancer prognosis involves predicting the course of the disease, the likelihood that it will spread and recur, and the likelihood that patients will survive. The clinical management of cancer patients will considerably benefit from the precision of cancer prognosis prediction. To better forecast cancer prognosis, modern statistical analysis and Deep learning techniques are being applied, as well as biomedical translational research being improved. In recent years, the processing capacity has significantly increased and the innovation of artificial insight, especially deep learning, has advanced quickly. Cancer is the leading reason for death in people. As a result, cancer detection is essential for early diagnosis and provides the best chance for treating cancer patients in a secure and efficient manner. It is, however, the trickiest way to increase the likelihood of the person surviving. RNA sequencing has significantly advanced in the last few decades and is now a crucial method for transcriptome profiling.

A guide to open science practices for animal research.

Translational biomedical research relies on animal experiments and provides the underlying proof of practice for clinical trials, which places an increased duty of care on translational researchers to derive the maximum possible output from every experiment performed. The implementation of open science practices has the potential to initiate a change in research culture that could improve the transparency and quality of translational research in general, as well as increasing the audience and scientific reach of published research. However, open science has become a buzzword in the scientific community that can often miss mark when it comes to practical implementation. In this Essay, we provide a guide to open science practices that can be applied throughout the research process, from study design, through data collection and analysis, to publication and dissemination, to help scientists improve the transparency and quality of their work. As open science practices continue to evolve, we also provide an online toolbox of resources that we will update continually.

Making translational value: Identifying ‘good targets’ for clinical research on gene editing and induced pluripotent stem cell technologies

Biomedical translational researchers aim to develop knowledge and techniques arising from research in the life sciences into clinical applications. Using the examples of induced pluripotent stem cells (iPSC) and gene editing, this paper examines how translational researchers identify and justify which particular conditions or patient populations make ‘good targets’ for translational research with particular technologies. Drawing on empirical data from qualitative interviews with academic and commercial researchers working on clinical translation of iPSC and gene editing in the UK, this study illustrates how particular combinations of technology and disease (for example iPSC-derived cells as a therapy for Parkinson's disease or gene editing for Cystic Fibrosis) were evaluated and justified as worth pursuing. The results show that translational researchers anticipate the ways in which their therapies-in-the making will be evaluated by other groups including regulators, physicians, patients, and bodies charged with health technology assessment. Each of these groups have their own understandings of what is valuable in a novel health technology and their own criteria for evaluation. As a result, translational researchers must supplement justifications that draw on scientific and industrial logics, with accounts that recognise other forms of worth, including market and civic registers of justification. These findings give an insight into the factors shaping contemporary biomedical translational research. The current regulatory and health technology adoption frameworks exert a strong influence, with elements such as ‘safety’ or ‘unmet need’ being common to most justification. However there was also sufficient flexibility to allow different competing definitions of what safety or unmet need might look like.

Analysis of demographic characteristics and disparities of enrollment in cancer clinical research at the U.S. National Cancer Institute (NCI): A 15-year experience.

6529 Background: Racial, gender, and socioeconomic disparities in cancer clinical trials participation have been well documented. The intramural program of the NCI in the National Institutes of Health (NIH) Bethesda campus conducts clinical research where participants are referred by their physician or self-referred, treated without charge and receive aid in travel expenses. We sought to describe the demographics of participants and identify potential disparities. Methods: We used the clinical research data repository of the NIH, Biomedical Translational Research Information System (BTRIS), to extract self-reported demographic data of enrollees in NCI protocols from 2005-2020. Comparisons were made with the U.S. population of patients with cancer as captured in the Surveillance, Epidemiology, and End Results (SEER) Program (2005-2018). The average household income per zip code was derived from the American Community Survey at five-year increments from 2011, 2015, and 2019. Results: We obtained data on 40,007 participants; 38,531 (95%) came from the U.S., of which 18,606 (48%) from DC, Maryland, or Virginia (DMV). Median age at consent was 54 years (range 2 – 89). Proportions of basic demographic attributes and corresponding values from SEER data were presented. Notably, Hispanic male, non-Hispanic female, and white female participants were under-represented in NCI clinical research compared to SEER data. African Americans were under-represented in the non-DMV population (5%) vs. the DMV (19%). Also, for patients from the DMV area, median income was consistently less for African American participants compared to White and Asian participants. Conclusions: The identified disparities may be partially due to the specific research activities available at the NCI, but strategies aiming at increasing the participation and representation of under-represented groups are required. [Table: see text]

Breast Cancer Therapy: The Potential Role of Mesenchymal Stem Cells in Translational Biomedical Research.

The potential role of mesenchymal stem cells (MSCs) in the treatment of metastatic cancers, including breast cancer, has been investigated for many years leading to encouraging results. The role of fat grafting and the related adipose-derived mesenchymal stem cells (AD-MSCs) has been detailed and described for breast reconstruction purposes confirming the safety of AD-MSCs. MSCs have great potential for delivering anticancer agents, suicide genes, and oncolytic viruses to tumors. Currently, many studies have focused on the products of MSCs, including extracellular vesicles (EVs), as a cell-free therapy. This work aimed to review and discuss the current knowledge on MSCs and their EVs in breast cancer therapy.

The Case for Modernizing Biomedical Research in Ireland through the Creation of an Irish 3Rs Centre.

Simple SummaryThe 3Rs principle indicates the measures to refine, reduce, and replace the use of animals in research. Thirteen out of the twenty-seven European countries already have 3Rs Centres. These support the development, acceptance, and/or implementation of methods that address the 3Rs principle at a research and educational level. To date, Ireland has no 3Rs centre. In this commentary, we present the reasons for Ireland to embrace the creation of a national Irish 3Rs Centre. We believe the centre will enable Ireland to be ready for the paradigm shift that is internationally occurring in biomedical research, towards the modernisation and replacement of tests carried out in animals when it is scientifically possible to do so.Since its publication, the 3Rs principle has provided a cornerstone for more ethical and humane biomedical and regulatory research. In Europe, the 3Rs principle has been incorporated into the European Directive 63/2010/EU, with the ultimate aim of fully replacing the procedures on live animals for scientific and educational purposes as soon as it is scientifically possible to do so. Thus, a critical shift in the discussion on animal use in biomedical and regulatory research is undergoing in Europe, a discussion where satisfying the “replacement” principle is becoming more and more defined as a scientific rather than ethical need. 3Rs Centres have been established in recent years across Europe. To date, Ireland has no 3Rs Centre, and the uptake of the 3Rs principle, and in particular of the “replacement” aspect, has been slow. In this Commentary, we present the Irish context of the use of animal models in biomedical and regulatory research, and urge for what, in the authors’ opinion, are the most critical actions that Ireland must undertake to align its biomedical (basic, applied and translational) research with the European 3Rs strategy.

The Survey of the Health of Wisconsin (SHOW) Program: An Infrastructure for Advancing Population Health.

IntroductionThe Survey of the Health of Wisconsin (SHOW) was established in 2008 by the University of Wisconsin (UW) School of Medicine and Public Health (SMPH) with the goals of (1) providing a timely and accurate picture of the health of the state residents; and (2) serving as an agile resource infrastructure for ancillary studies. Today, the SHOW program continues to serve as a unique and vital population health research infrastructure for advancing public health.MethodsSHOW currently includes 5,846 adult and 980 minor participants recruited between 2008 and 2019 in four primary waves. WAVE I (2008–2013) includes annual statewide representative samples of 3,380 adults ages 21 to 74 years. WAVE II (2014–2016) is a triannual statewide sample of 1,957 adults (age ≥18 years) and 645 children (age 0–17). WAVE III (2017) consists of follow-up of 725 adults from the WAVE I and baseline surveys of 222 children in selected households. WAVEs II and III include stool samples collected as part of an ancillary study in a subset of 784 individuals. WAVE IV consists of 517 adults and 113 children recruited from traditionally under-represented populations in biomedical research including African Americans and Hispanics in Milwaukee, Wisconsin.Findings to DateThe SHOW resource provides unique spatially granular and timely data to examine the intersectionality of multiple social determinants and population health. SHOW includes a large biorepository and extensive health data collected in a geographically diverse urban and rural population. Over 60 studies have been published covering a broad range of topics including, urban and rural disparities in cardio-metabolic disease and cancer, objective physical activity, sleep, green-space and mental health, transcriptomics, the gut microbiome, antibiotic resistance, air pollution, concentrated animal feeding operations and heavy metal exposures.DiscussionThe SHOW cohort and resource is available for continued follow-up and ancillary studies including longitudinal public health monitoring, translational biomedical research, environmental health, aging, microbiome and COVID-19 research.

Systematic Investigation of the Effects of Multiple SV40 Nuclear Localization Signal Fusion on the Genome Editing Activity of Purified SpCas9.

The emergence of CRISPR-Cas9 technology has revolutionized both basic and translational biomedical research. For Cas9 nuclease to exert genome editing activity, nuclear localization signal (NLS) derived from simian virus 40 (SV40) T antigen is commonly installed as genetic fusion to direct the intracellular Cas9 proteins to the nucleus of cells. Notably, previous studies have shown that multiple SV40 NLS fusion can improve the targeting activity of Cas9-derived genome-editing and base-editing tools. In addition, the multi-NLS fusion can increase the intracellular activity of Cas9 in the forms of both constitutive expression and directly delivered Cas9-guide RNA ribonucleoprotein (RNP) complex. However, the relationship between NLS fusion and intracellular Cas9 activity has not been fully understood, including the dependency of activity on the number or organization of NLS fusion. In the present study, we constructed and purified a set of Streptococcus pyogenes Cas9 (SpCas9) variants containing one to four NLS repeats at the N- or C-terminus of the proteins and systematically analyzed the effects of multi-NLS fusion on the activity of SpCas9 RNPs. It was found that multi-NLS fusion could improve the intracellular activity as lipofected or nucleofected Cas9 RNPs. Importantly, multi-NLS fusion could enhance the genome-editing activity of SpCas9 RNPs in primary and stem/progenitor cells and mouse embryos.

Biochemical and Hematologic Reference Intervals for Anesthetized, Female, Juvenile Yorkshire Swine.

Swine are widely used in biomedical research, translational research, xenotransplantation, and agriculture. For these uses, physiologic reference intervals are extremely important for assessing the health status of the swine and diagnosing disease. However, few biochemical and hematologic reference intervals that comply with guidelines from the Clinical and Laboratory Standards Institute and the American Society for Veterinary Clinical Pathology are available for swine. These guidelines state that reference intervals should be determined by using 120 subjects or more. The aim of this study was to generate hematologic and biochemical reference intervals for female, juvenile Yorkshire swine (Sus scrofa domesticus) and to compare these values with those for humans and baboons (Papio hamadryas). Blood samples were collected from the femoral artery or vein of female, juvenile Yorkshire swine, and standard hematologic and biochemical parameters were analyzed in multiple studies. Hematologic and biochemical reference intervals were calculated for arterial blood samples from Yorkshire swine (n = 121 to 124); human and baboon reference intervals were obtained from the literature. Arterial reference intervals for Yorkshire swine differed significantly from those for humans and baboons in all commonly measured parameters except platelet count, which did not differ significantly from the human value, and glucose, which was not significantly different from the baboon value. These data provide valuable information for investigators using female, juvenile Yorkshire swine for biomedical re- search, as disease models, and in xenotransplantation studies as well as useful physiologic information for veterinarians and livestock producers. Our findings highlight the need for caution when comparing data and study outcomes between species.

Understanding sex differences in zebrafish pain- and fear-related behaviors

Sex is an important variable in translational biomedical research. While overt sex differences have been reported for pain and fear-like behaviors in humans and rodents, these differences in other popular model organisms, such as zebrafish, remain poorly understood. Here, we evaluate potential sex differences in zebrafish behavioral responses to pain (intraperitoneal administration of 5% acetic acid) and fear stimuli (exposure to alarm substance). Overall, both male and female zebrafish exposed to pain (acetic acid injection) show lesser distance traveled, fewer top entries and more writhing-like pain-related behavior vs. controls, whereas female fish more robustly (than males) altered some other pain-like behaviors (e.g., increasing freezing episodes and time in top) in this model. In contrast, zebrafish of both sexes responded equally strongly to fear evoked by acute alarm substance exposure. Collectively, these findings emphasize the growing importance of studying sex differences in zebrafish behavioral and pain models.

Iron Status Is Inversely Correlated with Circulating Red Cells in Patients with Sickle Cell Anemia

▪Introduction: Sickle cell anemia (SCA, HbSS) arises from the presence of sickle hemoglobin (HbS, a consequence of a single base mutation in HBB gene). Upon deoxygenation, deoxy-HbS polymerizes with detrimental effects on shape, function, and lifespan of the erythrocytes. Because the delay time of sickling is inversely proportional to the mean cellular hemoglobin concentration (MCHC), decreasing the concentration of HbS is an attractive therapeutic approach. We recently demonstrated that dietary iron restriction decreases MCHC with a concomitant improvement in hematologic parameters and a decreased propensity to sickle in the Townes murine model of sickle cell anemia. Data describing the relationship between iron status, MCHC, and hematologic parameters in SCA patients are limited to isolated case series that do not account for alpha thalassemia, a known modifier of SCA severity. Based on our observations in mice, we hypothesize that decreased iron availability is associated with improved hematologic parameters in SCA patients in the absence of alpha thalassemia. We investigated the relationship between MCHC and red cell count, as well as transferrin saturation and red cell count, in healthy volunteers and sickle cell patients without alpha thalassemia.Methods: A database consisting of 795 patients with sickle cell disease (all genotypes) attending the National Institutes of Health between 2001 and 2015 was queried (see table). Data were filtered for sickle genotype, whether on hydroxyurea therapy, transfusion status (HbA), alpha thalassemia genotype, and relevant hematologic and/or iron panels obtained on the same visit. Data from healthy volunteers were obtained from the National Institutes of Health Biomedical Translational Research Information System (BTRIS) database filtering for those without detectable HbS that had hematologic and iron panel data obtained on the same visit. Correlations were analyzed by the Pearson correlation coefficient, r. p-values < 0.05 were considered statistically significant.Results: Analyses were performed on patients with HbSS without detectable HbA that were not taking hydroxyurea and did not have alpha thalassemia. Thirty-nine of these patients had MCHC and red cell counts obtained on the same visit. Data indicate that MCHC is inversely correlated with red cell count in these patients (r=-0.423; p = 0.007). Importantly, no significant relationship between MCHC and red cell count was observed in healthy volunteers (n=40; r=-0.125; p =0.438). Transferrin saturation is directly correlated with circulating red cells in healthy volunteers (n=65; r=0.359; p=0.0033). However, in patients with sickle cell disease transferrin saturation has an inverse relationship with red cell count (n=53; r=-0.3131; p=0.0225).Conclusions: Transferrin saturation and MCHC have are inversely correlated with circulating red cell counts in patients with sickle cell disease. These results support the concept that decreased iron availability improves hematologic parameters in sickle cell disease. Future studies will focus on pharmacologic means to achieve this end. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Using bibliometrics to evaluate outcomes and influence of translational biomedical research centers.

Federal grant funding to support infrastructure development of translational biomedical research centers is a form of public health intervention. Establishing rigorous methods for measuring center success and outcomes is essential to justify continued funding. Bibliometric data compiled from a 5-year funding cycle of neurodegeneration and translational neuroscience research center were analyzed using the package bibliometrix for open-source software R and the NIH-developed research tool iCite. The research team and their collaborators (n = 485) produced 157 grant-citing publications from 2015-2020. The science was produced by small research teams clustered around three main communities of topics: Alzheimer's Disease, brain imaging, and neuropsychological testing in the elderly. Using the relative citation ratio, the publications produced by the research team were found to be influential when compared to other R01-funded publications. Recent developments in bibliometric analysis expand beyond traditional measurement capabilities to better understand the characteristics, outcomes, and influences of research teams. These findings can be used to inform researchers and institutions about research team composition, productivity, and success. Measures of research influence may be used to justify return on investment to funders.

Radiomics in Head and Neck Cancers Radiotherapy. Promises and Challenges

Radiomics, a subdomain of artificial intelligence, consists in extracting a large volume of data from all medical imaging techniques and correlating them with clinical data in order to build predictive and prognostic models. Radiomics is related to radiogenomics that correlates genetic mutations and molecular and biological characteristics of tissues with information extracted from medical imaging. Both are state-of-the-art fields of translational biomedical research. The ability to predict early patient survival and response to treatment, but also the capacity to identify tumor subtypes non-invasively, could make radiomics a key player with an essential role in personalized oncology. In head and neck cancer radiotherapy, radiomic algorithms can predict not only the response to radiochemotherapy or induction chemotherapy but also the need for planning through adaptive radiotherapy (ART). Radiomics can also predict the risk of severe toxicities, especially that of xerostomia. Given the benefit that a de-escalation of treatment can bring in selected cases to improve the quality of life, radiomics is expected to be part of the therapeutic decision for head and neck cancers in the near future, and may help identify cases where de-escalation of multimodal therapy will not jeopardize the therapeutic benefit.

Promoting FAIR Data Through Community-driven Agile Design: the Open Data Commons for Spinal Cord Injury (odc-sci.org)

The past decade has seen accelerating movement from data protectionism in publishing toward open data sharing to improve reproducibility and translation of biomedical research. Developing data sharing infrastructures to meet these new demands remains a challenge. One model for data sharing involves simply attaching data, irrespective of its type, to publisher websites or general use repositories. However, some argue this creates a ‘data dump’ that does not promote the goals of making data Findable, Accessible, Interoperable and Reusable (FAIR). Specialized data sharing communities offer an alternative model where data are curated by domain experts to make it both open and FAIR. We report on our experiences developing one such data-sharing ecosystem focusing on ‘long-tail’ preclinical data, the Open Data Commons for Spinal Cord Injury (odc-sci.org). ODC-SCI was developed with community-based agile design requirements directly pulled from a series of workshops with multiple stakeholders (researchers, consumers, non-profit funders, governmental agencies, journals, and industry members). ODC-SCI focuses on heterogeneous tabular data collected by preclinical researchers including bio-behaviour, histopathology findings and molecular endpoints. This has led to an example of a specialized neurocommons that is well-embraced by the community it aims to serve. In the present paper, we provide a review of the community-based design template and describe the adoption by the community including a high-level review of current data assets, publicly released datasets, and web analytics. Although odc-sci.org is in its late beta stage of development, it represents a successful example of a specialized data commons that may serve as a model for other fields.

Comparative annotation of porcine bone marrow cell types using scRNAseq

Abstract Pigs are important as a biomedical model and for worldwide meat production. Therefore, a better understanding of porcine hematopoiesis and immune system development could have a wide impact on human disease research and food production. The bone marrow represents a unique environment that is the major site of differentiation of stem cells and progenitors into mature blood cells. We performed scRNA-seq on bone marrow from healthy adult pigs and sequenced a total of 3,576 cells. After QC, 2,707 cells and 16,432 genes were used for downstream analysis. The porcine bone marrow cells were grouped into 15 distinct clusters and these relationships were mapped using non-linear dimensional reduction. To find markers that defined each cluster, we found a total of 3,335 differentially expressed genes using the non-parameteric Wilcoxon rank sum test. Further, pairwise differential expression testing and random forest models were used to evaluate the similarity between clusters and further define the unique features of each cluster. Using these defined markers, we were able to annotate the cell types for each cluster. The clusters represented a diverse set of cell types including myeloid and lymphoid cells, myeloid and lymphoid cell progenitors, and platelet and red blood cell progenitors. We validated these annotations using publicly available datasets including the CITE-seq database of human bone marrow mononuclear cells (Stuart et al., 2019). We also compared the expression profiles between similarly annotated cell types between human and pig. Our novel annotation of porcine bone marrow cells will be a resource for the study of porcine hematopoiesis and immune system development and help inform human translational biomedical research.

Cas9-expressing chickens and pigs as resources for genome editing in livestock

Genetically modified animals continue to provide important insights into the molecular basis of health and disease. Research has focused mostly on genetically modified mice, although other species like pigs resemble the human physiology more closely. In addition, cross-species comparisons with phylogenetically distant species such as chickens provide powerful insights into fundamental biological and biomedical processes. One of the most versatile genetic methods applicable across species is CRISPR-Cas9. Here, we report the generation of transgenic chickens and pigs that constitutively express Cas9 in all organs. These animals are healthy and fertile. Functionality of Cas9 was confirmed in both species for a number of different target genes, for a variety of cell types and in vivo by targeted gene disruption in lymphocytes and the developing brain, and by precise excision of a 12.7-kb DNA fragment in the heart. The Cas9 transgenic animals will provide a powerful resource for in vivo genome editing for both agricultural and translational biomedical research, and will facilitate reverse genetics as well as cross-species comparisons.

Gene Editing in Rabbits: Unique Opportunities for Translational Biomedical Research.

The rabbit is a classic animal model for biomedical research, but the production of gene targeted transgenic rabbits had been extremely challenging until the recent advent of gene editing tools. More than fifty gene knockout or knock-in rabbit models have been reported in the past decade. Gene edited (GE) rabbit models, compared to their counterpart mouse models, may offer unique opportunities in translational biomedical research attributed primarily to their relatively large size and long lifespan. More importantly, GE rabbit models have been found to mimic several disease pathologies better than their mouse counterparts particularly in fields focused on genetically inherited diseases, cardiovascular diseases, ocular diseases, and others. In this review we present selected examples of research areas where GE rabbit models are expected to make immediate contributions to the understanding of the pathophysiology of human disease, and support the development of novel therapeutics.

Recommendations of National symposium on “Integrative Medicines and Health: From Basic to Translational Research”

Translational Biomedical Research Society (TBRS), India founded in the year 2019, strives to promote communication and integration of traditional and standard healthcare systems through bridging the gap between Ancient Science and Modern Medicine. Continuing in this direction, TBRS took the lead to organize a national symposium entitled “Integrative Medicines and Health: From Basic to Translational Research” during December 4-6, 2020 as TBRS-2020. The symposium was jointly organized by Centre of Biomedical Research (CBMR), Lucknow and Faculty of Biosciences Institute of Biosciences and Technology, Shri Ramswaroop Memorial University (SRMU), Barabanki, UP in online mode. This meeting was a successful endeavor promoting integration of Natural Healthcare systems and Modern medical care systems for the benefit of human society. The theme of the symposium revolved around integrative health care so that to bring conventional and complementary approaches together with mainstream medical care systems. The use of integrative approaches to healthcare management is growing rapidly, especially, within critical care settings and researchers are currently exploring the potential benefits of integrative medicines in a variety of situations, including relief of symptoms in cancer patients and survivors, and programs to promote healthy life style and healthy behaviors.

Cardiovascular bioimaging of nitric oxide: Achievements, challenges, and the future.

Nitric oxide (NO) is a ubiquitous, volatile, cellular signaling molecule that operates across a wide physiological concentration range (pM-µM) in different tissues. It is a highly diffusible messenger and intermediate in various metabolic pathways. NO plays a pivotal role in maintaining optimum cardiovascular function, particularly by regulating vascular tone and blood flow. This review highlights the need for accurate, real-time bioimaging of NO in clinical diagnostic, therapeutic, monitoring, and theranostic applications within the cardiovascular system. We summarize electrochemical, optical, and nanoscale sensors that allow measurement and imaging of NO, both directly and indirectly via surrogate measurements. The physical properties of NO render it difficult to accurately measure in tissues using direct methods. There are also significant limitations associated with the NO metabolites used as surrogates to indirectly estimate NO levels. All these factors added to significant variability in the measurement of NO using available methodology have led to a lack of sensors and imaging techniques of clinical applicability in relevant vascular pathologies such as atherosclerosis and ischemic heart disease. Challenges in applying current methods to biomedical and clinical translational research, including the wide physiological range of NO and limitations due to the characteristics and toxicity of the sensors are discussed, as are potential targets and modifications for future studies. The development of biocompatible nanoscale sensors for use in combination with existing clinical imaging modalities provides a feasible opportunity for bioimaging NO within the cardiovascular system.