Richard M Caprioli

Vanderbilt University, Nashville, Michigan, United States

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Publications (369)1728.35 Total impact

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    ABSTRACT: The present study was designed to demonstrate the potential of an optimized histology directed protein identification combined with imaging mass spectrometry technology to reveal and identify molecules associated to ectopic calcification in human tissue. As a proof of concept, mineralized and non-mineralized areas were compared within the same dermal tissue obtained from a patient affected by Pseudoxanthoma elasticum, a genetic disorder characterized by calcification only at specific sites of soft connective tissues. Data have been technically validated on a contralateral dermal tissue from the same subject and compared with those from control healthy skin. Results demonstrate that this approach 1) significantly reduces the effects generated by techniques that, disrupting tissue organization, blend data from affected and unaffected areas; 2) demonstrates that, abolishing differences due to inter-individual variability, mineralized and non-mineralized areas within the same sample have a specific protein profile and have a different distribution of molecules; 3) avoiding the bias of focusing on already known molecules, reveals a number of proteins that have been never related to the disease nor to the calcification process, thus paving the way for the selection of new molecules to be validated as pathogenic or as potential pharmacological targets.
    Bone 01/2015; 24. · 4.46 Impact Factor
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    ABSTRACT: Imaging mass spectrometry (IMS) was employed for the analysis of frozen skin biopsies to investigate the differences between stage IV pressure ulcers that remain stalled, stagnant and unhealed versus those exhibiting clinical and histological signs of improvement. Our data reveal a rich diversity of proteins that are dynamically modulated and we selectively highlight a family of calcium binding proteins (S-100 molecules) including calcyclin (S100-A6), calgranulins A (S100-A8) and B (S100-A9), and calgizzarin (S100-A11). IMS allowed us to target 3 discrete regions of interest: the wound bed, adjacent dermis, and hypertrophic epidermis. Plots derived using unsupervised principal component analysis of the global protein signatures within these 3 spatial niches indicate that these data from wound signatures have potential as a prognostic tool since they appear to delineate wounds that are favorably responding to therapeutic interventions versus those that remain stagnant or intractable in their healing status. Our discovery based approach with IMS augments current knowledge of the molecular signatures within pressure ulcers while providing a rationale for a focused examination of the role of calcium modulators within the context of impaired wound healing.
    Journal of Proteome Research 12/2014; · 5.06 Impact Factor
  • Domenico Taverna, Jeremy L Norris, Richard M Caprioli
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    ABSTRACT: This study presents an on-tissue proteolytic digestion and peptide extraction method using microwave irradiation for in situ analysis of proteins from spatially defined regions of a tissue section. The methodology utilizes hydrogel discs (1 mm diameter) embedded with trypsin solution. The hydrogel discs are applied to a tissue section, directing enzymatic digestion to a spatially confined area of the tissue. By applying microwave radiation, protein digestion is performed in 2 minutes on-tissue, and the extracted peptides are then analyzed by MALDI MS and LC-MS/MS. The reliability and reproducibility of the microwave-assisted hydrogel mediated on-tissue digestion is demonstrated by the comparison with other on-tissue digestion strategies, including comparisons with conventional heating and in-solution digestion. LC-MS/MS data were evaluated considering the number of identified proteins as well as the number of protein groups and distinct peptides. The results of this study demonstrate that a rapid and reliable protein digestion can be performed on a single thin tissue section while preserving the tissue architecture, and the resulting peptides can be extracted in sufficient abundance to permit analysis using LC-MS/MS. This approach will be most useful for samples that have limited availability but are needed for multiple analyses, especially for the correlation of proteomics data with histology and immunohistochemistry.
    Analytical Chemistry 11/2014; · 5.83 Impact Factor
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    ABSTRACT: Graphical abstract Figure optionsDownload full-size imageDownload as PowerPoint slide
    EuPA Open Proteomics. 09/2014;
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    ABSTRACT: Imaging mass spectrometry (IMS) has become a prime tool for studying the distribution of biomolecules in tissue. Although IMS data sets can become very large, computational methods have made it practically feasible to search these experiments for relevant findings. However, these methods lack access to an important source of information that many human interpretations rely upon: anatomical insight. In this work, we address this need by (1) integrating a curated anatomical data source with an empirically acquired IMS data source, establishing an algorithm-accessible link between them and (2) demonstrating the potential of such an IMS-anatomical atlas link by applying it toward automated anatomical interpretation of ion distributions in tissue. The concept is demonstrated in mouse brain tissue, using the Allen Mouse Brain Atlas as the curated anatomical data source that is linked to MALDI-based IMS experiments. We first develop a method to spatially map the anatomical atlas to the IMS data sets using nonrigid registration techniques. Once a mapping is established, a second computational method, called correlation-based querying, gives an elementary demonstration of the link by delivering basic insight into relationships between ion images and anatomical structures. Finally, a third algorithm moves further beyond both registration and correlation by providing automated anatomical interpretation of ion images. This task is approached as an optimization problem that deconstructs ion distributions as combinations of known anatomical structures. We demonstrate that establishing a link between an IMS experiment and an anatomical atlas enables automated anatomical annotation, which can serve as an important accelerator both for human and machine-guided exploration of IMS experiments.
    Analytical Chemistry 08/2014; · 5.83 Impact Factor
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    ABSTRACT: Imaging Mass Spectrometry is increasingly being applied to the study of small endogenous compounds, including metabolites, neurotransmitters, lipids and other compounds as well. However, due to the high degree of structural homology and the lack of true "blank" samples, generation of images of unequivocal molecular identity is challenging. In this special feature perspective article, Richard Caprioli and colleagues at Vanderbilt University Medical Center discuss these challenges and describe an analytical strategy that combines a number of advanced instrumental methods to identify and confirm the accurate spatial localization of select amino acids and amine-containing metabolites. By combining derivatization, MS(n) , and accurate mass, followed by confirmation via HPLC-MS, they are able to demonstrate the localization of several endogenous metabolites in biological tissue specimens.
    Biological Mass Spectrometry 08/2014; 49(8). · 2.71 Impact Factor
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    ABSTRACT: Imaging mass spectrometry (IMS) studies increasingly focus on endogenous small molecular weight metabolites and consequently bring special analytical challenges. Since analytical tissue blanks do not exist for endogenous metabolites, careful consideration must be given to confirm molecular identity. Here, we present approaches for the improvement in detection of endogenous amine metabolites such as amino acids and neurotransmitters in tissues through chemical derivatization and matrix-assisted laser desorption/ionization (MALDI) IMS. Chemical derivatization with 4-hydroxy-3-methoxycinnamaldehyde (CA) was used to improve sensitivity and specificity. CA was applied to the tissue via MALDI sample targets precoated with a mixture of derivatization reagent and ferulic acid as a MALDI matrix. Spatial distributions of chemically derivatized endogenous metabolites in tissue were determined by high-mass resolution and MSn IMS. We highlight an analytical strategy for metabolite validation whereby tissue extracts are analyzed by high-performance liquid chromatography (HPLC)-MS/MS to unambiguously identify metabolites and distinguish them from isobaric compounds. Copyright © 2014 John Wiley & Sons, Ltd.
    Biological Mass Spectrometry 08/2014; 49(8). · 2.71 Impact Factor
  • Jessica L Moore, Richard M Caprioli, Eric P Skaar
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    ABSTRACT: Matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) has been successfully applied to the field of microbial pathogenesis with promising results, principally in diagnostic microbiology to rapidly identify bacteria based on the molecular profiles of small cell populations. Direct profiling of molecules from serum and tissue samples by MALDI MS provides a means to study the pathogen-host interaction and to discover potential markers of infection. Systematic molecular profiling across tissue sections represents a new imaging modality, enabling regiospecific molecular measurements to be made in situ, in both two-dimensional and three-dimensional analyses. Herein, we briefly summarize work that employs MALDI MS to study the pathogenesis of microbial infection.
    Current Opinion in Microbiology 07/2014; 19C:45-51. · 7.22 Impact Factor
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    Junhai Yang, Andre Zavalin, Richard Caprioli
    62th ASMS Conference on Mass Spectrometry and Allied Topics; 06/2014
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    62th ASMS Conference on Mass Spectrometry and Allied Topics; 06/2014
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    ABSTRACT: Diabetic nephropathy (DN) is a major life-threatening complication of diabetes. Renal lesions affect glomeruli and tubules but the pathogenesis is not completely understood. Phospholipids and glycolipids are molecules that carry out multiple cell functions in norm and disease and their role in DN pathogenesis is unknown. We employed high spatial resolution matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI IMS) to determine lipid changes in kidneys of eNOS(-/-) db/db mice, a robust model of DN. Phospholipid and glycolipid structures, localization patterns, and relative tissue levels were determined in individual renal glomeruli and tubules without disturbing tissue morphology. Significant increase in the levels of specific glomerular and tubular lipid species from four different classes, i.e. gangliosides, sulfoglycosphingolipids, lysophospholipids, and phosphatidylethanolamines was detected in diabetic kidneys compared to non-diabetic controls. Inhibition of non-enzymatic oxidative and glycoxidative pathways attenuated the increase in lipid levels and ameliorated renal pathology, even though blood glucose levels remained unchanged. Our data demonstrate that the levels of specific phospho- and glycolipids in glomeruli and/or tubules are associated with diabetic renal pathology. We suggest that hyperglycemia-induced DN pathogenic mechanisms require intermediate oxidative steps that involve specific phospholipid and glycolipid species.
    The Journal of Lipid Research 05/2014; · 4.73 Impact Factor
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    ABSTRACT: Matrix assisted laser desorption ionization imaging mass spectrometry (MALDI IMS) has the ability to provide an enormous amount of information on the abundances and spatial distributions of molecules within biological tissues. The rapid progress in the development of this technology significantly improves our ability to analyze smaller and smaller areas and features within tissues. The mammalian eye has evolved over millions of years to become an essential asset for survival, providing important sensory input of an organism's surroundings. The highly complex sensory retina of the eye is comprised of numerous cell types organized into specific layers with varying dimensions, the thinnest of which is the 10 μm retinal pigment epithelium (RPE). This single cell layer and the photoreceptor layer contain the complex biochemical machinery required to convert photons of light into electrical signals that are transported to the brain by axons of retinal ganglion cells. Diseases of the retina, including age-related macular degeneration (AMD), retinitis pigmentosa, and diabetic retinopathy, occur when the functions of these cells are interrupted by molecular processes that are not fully understood. In this report, we demonstrate the use of high spatial resolution MALDI IMS and FT-ICR tandem mass spectrometry in the Abca4 (-/-) knockout mouse model of Stargardt disease, a juvenile onset form of macular degeneration. The spatial distributions and identity of lipid and retinoid metabolites are shown to be unique to specific retinal cell layers.
    Journal of the American Society for Mass Spectrometry 05/2014; 25(8). · 3.59 Impact Factor
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    ABSTRACT: We identified Acyl-Coenzyme A Binding Protein (ACBP) as part of a proteomic signature predicting the risk of having lung cancer. Because ACBP is known to regulate beta oxidation (β-oxidation), which in turn controls cellular proliferation, we hypothesized that ACBP contributes to regulation of cellular proliferation and survival of non-small cell lung cancer (NSCLC) by modulating β-oxidation. We utilized matrix assisted laser desorption ionization- imaging mass spectrometry (MALDI-IMS) and immunohistochemistry (IHC) to confirm ACBP's tissue localization in pre-invasive and invasive NSCLCs. We correlated ACBP gene expression levels in NSCLC with clinical outcomes. In loss of function studies, we tested the effect of the downregulation of ACBP on cellular proliferation and apoptosis in normal bronchial and NSCLC cell lines. Using tritiated-palmitate (3H-palmitate), we measured β-oxidation levels and tested the effect of etomoxir, a β-oxidation inhibitor, on proliferation and apoptosis. MALDI-IMS and IHC analysis confirmed that ACBP is overexpressed in preinvasive and invasive lung cancers. High ACBP gene expression levels in NSCLCs correlated with worse survival (HR = 1.73). We observed a 40% decrease in β-oxidation and concordant decreases in proliferation and increases in apoptosis in ACBP depleted NSCLC cells as compared to bronchial airway epithelial cells. Inhibition of β-oxidation by etomoxir in ACBP overexpressing cells produced dose-dependent decrease in proliferation, and increase in apoptosis (p=0.01 and p <0.001 respectively). These data suggest a role for ACBP in controlling lung cancer progression by regulating β-oxidation.
    Cancer Prevention Research 05/2014; · 4.89 Impact Factor
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    Junhai Yang, Richard M. Caprioli
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    ABSTRACT: We have developed matrix pre-coated targets for imaging proteins in thin tissue sections by matrix-assisted laser desorption/ionization mass spectrometry. Gold covered microscope slides were coated with sinapinic acid (SA) in batches in advance and were shown to be stable for over 6 months when kept in the dark. The sample preparation protocol using these SA pre-coated targets involves treatment with diisopropylethylamine (DIEA)-H2O vapor, transforming the matrix layer to a viscous ionic liquid. This SA-DIEA ionic liquid layer extracts proteins and other analytes from tissue sections that are thaw mounted to this target. DIEA is removed by the immersion of the target into diluted acetic acid, allowing SA to co-crystallize with extracted analytes directly on the target. Ion images (3–70 kDa) of sections of mouse brain and rat kidney at spatial resolution down to 10 µm were obtained. Use of pre-coated slides greatly reduces sample preparation time for matrix-assisted laser desorption/ionization imaging while providing high throughput, low cost and high spatial resolution images. Copyright © 2014 John Wiley & Sons, Ltd.
    Biological Mass Spectrometry 05/2014; 49(5). · 2.71 Impact Factor
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    Molecular Psychiatry 05/2014; 19(5):529. · 15.15 Impact Factor
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    ABSTRACT: We have investigated the use of a Gaussian beam laser for MALDI Imaging Mass Spectrometry to provide a precisely defined laser spot of 5 μm diameter on target using a commercial MALDI TOF instrument originally designed to produce a 20 μm diameter laser beam spot at its smallest setting. A Gaussian beam laser was installed in the instrument in combination with an aspheric focusing lens. This ion source produced sharp ion images at 5 μm spatial resolution with signals of high intensity as shown for images from thin tissue sections of mouse brain.
    Journal of the American Society for Mass Spectrometry 04/2014; · 3.59 Impact Factor
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    ABSTRACT: Wilms tumor (WT) is the most common childhood kidney cancer worldwide and arises in children of black African ancestry with greater frequency and severity than other race groups. A biologic basis for this pediatric cancer disparity has not been previously determined. We hypothesized that unique molecular fingerprints might underlie the variable incidence and distinct disease characteristics of WT observed between race groups. To evaluate molecular disparities between WTs of different race groups, the Children's Oncology Group provided 80 favorable histology specimens divided evenly between black and white patients and matched for disease characteristics. As a surrogate of black sub-Saharan African patients, we also analyzed 18 Kenyan WT specimens. Tissues were probed for peptide profiles using matrix-assisted laser desorption ionization time of flight imaging mass spectrometry. To control for histologic variability within and between specimens, cellular regions were analyzed separately as triphasic (containing blastema, epithelia, and stroma), blastema only, and stroma only. Data were queried using ClinProTools and statistically analyzed. Peptide profiles, detected in triphasic WT regions, recognized race with good accuracy, which increased for blastema- or stroma-only regions. Peptide profiles from North American WTs differed between black and white race groups but were far more similar in composition than Kenyan specimens. Individual peptides were identified that also associated with WT patient and disease characteristics (eg, treatment failure and stage). Statistically significant peptide fragments were used to sequence proteins, revealing specific cellular signaling pathways and candidate drug targets. Wilms tumor specimens arising among different race groups show unique molecular fingerprints that could explain disparate incidences and biologic behavior and that could reveal novel therapeutic targets.
    Journal of the American College of Surgeons 04/2014; 218(4):707-20. · 4.45 Impact Factor
  • Megan M Gessel, Jeremy L Norris, Richard M Caprioli
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    ABSTRACT: Matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI IMS) combines the sensitivity and selectivity of mass spectrometry with spatial analysis to provide a new dimension for histological analyses to provide unbiased visualization of the arrangement of biomolecules in tissue. As such, MALDI IMS has the capability to become a powerful new molecular technology for the biological and clinical sciences. In this review, we briefly describe several applications of MALDI IMS covering a range of molecular weights, from drugs to proteins. Current limitations and challenges are discussed along with recent developments to address these issues.
    Journal of proteomics 03/2014; · 5.07 Impact Factor
  • Cancer Epidemiology Biomarkers & Prevention 03/2014; 21(10_Supplement):B71-B71. · 4.32 Impact Factor
  • Richard M Caprioli
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    ABSTRACT: The molecular expression from genomic, proteomic, and metabolomic processes ongoing in living cells is enormously complex, continuously challenging our ability to measure and understand their integrated totality. Important biological studies and advances in the understanding of the biochemistry and biology of living cells have most often been preceded by innovations in the technology used to probe cells, tissues, and animals that then facilitate new insightful observations. A great deal has been learned over the decades about individual enzymes and pathways from isolated samples and extracts from a wide variety of bacterial and animal sources and much progress has been made toward integrating these findings into a framework that describes the underlying biology. This article is protected by copyright. All rights reserved.
    Proteomics 02/2014; 14(7-8). · 3.97 Impact Factor

Publication Stats

12k Citations
1,728.35 Total Impact Points


  • 1998–2014
    • Vanderbilt University
      • • Mass Spectrometry Research Center
      • • Department of Biochemistry
      • • Department of Neurological Surgery
      • • Medical Center
      • • Division of Allergy, Pulmonary and Critical Care
      • • Department of Chemistry
      Nashville, Michigan, United States
  • 2012
    • Cairo University
      • Faculty of Pharmacy
      Cairo, Muhafazat al Qahirah, Egypt
    • University of Leipzig
      • Medizinische Fakultät
      Leipzig, Saxony, Germany
  • 2011
    • Università degli Studi di Milano-Bicocca
      • Department of Health Science
      Milano, Lombardy, Italy
    • Università della Calabria
      Rende, Calabria, Italy
    • Yale University
      New Haven, Connecticut, United States
  • 2010
    • University of Colorado
      • Division of Endocrinology, Metabolism and Diabetes
      Denver, CO, United States
    • David H. Murdock Research Institute
      North Carolina, United States
  • 2009
    • American Society for Mass Spectrometry
      Nashville, Tennessee, United States
  • 2003–2007
    • Uppsala University
      • Department of Pharmaceutical Biosciences
      Uppsala, Uppsala, Sweden
    • Gateway-Vanderbilt Cancer Treatment Center
      Clarksville, Tennessee, United States
  • 2006
    • New York University
      • Department of Pathology
      New York City, NY, United States
  • 1978–2005
    • University of Texas Medical School
      • • Department of Biochemistry and Molecular Biology
      • • Department of Neurobiology and Anatomy
      Houston, Texas, United States
  • 1969–2005
    • Purdue University
      • Department of Chemistry
      West Lafayette, IN, United States
  • 2001
    • Oak Ridge National Laboratory
      Oak Ridge, Florida, United States
    • Chennai Institute Of Technology
      Chennai, Tamil Nādu, India
  • 1983
    • University of Texas MD Anderson Cancer Center
      • Department of Pathology
      Houston, Texas, United States
  • 1982
    • University of Texas Health Science Center at Houston
      Houston, Texas, United States