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  1. Article: Proteogenomic convergence for understanding cancer pathways and networks.

    Boja, Emily S / Rodriguez, Henry

    Clinical proteomics

    2014  Volume 11, Issue 1, Page(s) 22

    Abstract: During the past several decades, the understanding of cancer at the molecular level has been primarily focused on mechanisms on how signaling molecules transform homeostatically balanced cells into malignant ones within an individual pathway. However, it ...

    Abstract During the past several decades, the understanding of cancer at the molecular level has been primarily focused on mechanisms on how signaling molecules transform homeostatically balanced cells into malignant ones within an individual pathway. However, it is becoming more apparent that pathways are dynamic and crosstalk at different control points of the signaling cascades, making the traditional linear signaling models inadequate to interpret complex biological systems. Recent technological advances in high throughput, deep sequencing for the human genomes and proteomic technologies to comprehensively characterize the human proteomes in conjunction with multiplexed targeted proteomic assays to measure panels of proteins involved in biologically relevant pathways have made significant progress in understanding cancer at the molecular level. It is undeniable that proteomic profiling of differentially expressed proteins under many perturbation conditions, or between normal and "diseased" states is important to capture a first glance at the overall proteomic landscape, which has been a main focus of proteomics research during the past 15-20 years. However, the research community is gradually shifting its heavy focus from that initial discovery step to protein target verification using multiplexed quantitative proteomic assays, capable of measuring changes in proteins and their interacting partners, isoforms, and post-translational modifications (PTMs) in response to stimuli in the context of signaling pathways and protein networks. With a critical link to genotypes (i.e., high throughput genomics and transcriptomics data), new and complementary information can be gleaned from multi-dimensional omics data to (1) assess the effect of genomic and transcriptomic aberrations on such complex molecular machinery in the context of cell signaling architectures associated with pathological diseases such as cancer (i.e., from genotype to proteotype to phenotype); and (2) target pathway- and network-driven changes and map the fluctuations of these functional units (proteins) responsible for cellular activities in response to perturbation in a spatiotemporal fashion to better understand cancer biology as a whole system.
    Language English
    Publishing date 2014-06-01
    Publishing country England
    Document type Journal Article ; Review
    ZDB-ID 2205154-5
    ISSN 1542-6416
    ISSN 1542-6416
    DOI 10.1186/1559-0275-11-22
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Mapping the KRAS proteoform landscape in colorectal cancer identifies truncated KRAS4B that decreases MAPK signaling.

    Adams, Lauren M / DeHart, Caroline J / Drown, Bryon S / Anderson, Lissa C / Bocik, William / Boja, Emily S / Hiltke, Tara M / Hendrickson, Christopher L / Rodriguez, Henry / Caldwell, Michael / Vafabakhsh, Reza / Kelleher, Neil L

    The Journal of biological chemistry

    2022  Volume 299, Issue 1, Page(s) 102768

    Abstract: The KRAS gene is one of the most frequently mutated oncogenes in human cancer and gives rise to two isoforms, KRAS4A and KRAS4B. KRAS post-translational modifications (PTMs) have the potential to influence downstream signaling. However, the relationship ... ...

    Abstract The KRAS gene is one of the most frequently mutated oncogenes in human cancer and gives rise to two isoforms, KRAS4A and KRAS4B. KRAS post-translational modifications (PTMs) have the potential to influence downstream signaling. However, the relationship between KRAS PTMs and oncogenic mutations remains unclear, and the extent of isoform-specific modification is unknown. Here, we present the first top-down proteomics study evaluating both KRAS4A and KRAS4B, resulting in 39 completely characterized proteoforms across colorectal cancer cell lines and primary tumor samples. We determined which KRAS PTMs are present, along with their relative abundance, and that proteoforms of KRAS4A versus KRAS4B are differentially modified. Moreover, we identified a subset of KRAS4B proteoforms lacking the C185 residue and associated C-terminal PTMs. By confocal microscopy, we confirmed that this truncated GFP-KRAS4B
    MeSH term(s) Humans ; Colorectal Neoplasms/genetics ; Protein Isoforms/genetics ; Protein Isoforms/metabolism ; Protein Processing, Post-Translational ; Proto-Oncogene Proteins p21(ras)/genetics ; Proto-Oncogene Proteins p21(ras)/metabolism ; Cell Line, Tumor ; Signal Transduction ; Proteomics ; Mitogen-Activated Protein Kinases/metabolism
    Chemical Substances KRAS protein, human ; Protein Isoforms ; Proto-Oncogene Proteins p21(ras) (EC 3.6.5.2) ; Mitogen-Activated Protein Kinases (EC 2.7.11.24)
    Language English
    Publishing date 2022-12-05
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 2997-x
    ISSN 1083-351X ; 0021-9258
    ISSN (online) 1083-351X
    ISSN 0021-9258
    DOI 10.1016/j.jbc.2022.102768
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Mass spectrometry-based targeted quantitative proteomics: achieving sensitive and reproducible detection of proteins.

    Boja, Emily S / Rodriguez, Henry

    Proteomics

    2012  Volume 12, Issue 8, Page(s) 1093–1110

    Abstract: Traditional shotgun proteomics used to detect a mixture of hundreds to thousands of proteins through mass spectrometric analysis, has been the standard approach in research to profile protein content in a biological sample which could lead to the ... ...

    Abstract Traditional shotgun proteomics used to detect a mixture of hundreds to thousands of proteins through mass spectrometric analysis, has been the standard approach in research to profile protein content in a biological sample which could lead to the discovery of new (and all) protein candidates with diagnostic, prognostic, and therapeutic values. In practice, this approach requires significant resources and time, and does not necessarily represent the goal of the researcher who would rather study a subset of such discovered proteins (including their variations or posttranslational modifications) under different biological conditions. In this context, targeted proteomics is playing an increasingly important role in the accurate measurement of protein targets in biological samples in the hope of elucidating the molecular mechanism of cellular function via the understanding of intricate protein networks and pathways. One such (targeted) approach, selected reaction monitoring (or multiple reaction monitoring) mass spectrometry (MRM-MS), offers the capability of measuring multiple proteins with higher sensitivity and throughput than shotgun proteomics. Developing and validating MRM-MS-based assays, however, is an extensive and iterative process, requiring a coordinated and collaborative effort by the scientific community through the sharing of publicly accessible data and datasets, bioinformatic tools, standard operating procedures, and well characterized reagents.
    MeSH term(s) Biomarkers/analysis ; Databases, Protein ; High-Throughput Screening Assays ; Humans ; Mass Spectrometry/instrumentation ; Mass Spectrometry/methods ; Mass Spectrometry/standards ; Protein Processing, Post-Translational ; Proteins/analysis ; Proteomics/instrumentation ; Proteomics/methods ; Proteomics/standards ; Reference Standards ; Reproducibility of Results ; Sensitivity and Specificity ; Software
    Chemical Substances Biomarkers ; Proteins
    Language English
    Publishing date 2012-04
    Publishing country Germany
    Document type Journal Article ; Review
    ZDB-ID 2032093-0
    ISSN 1615-9861 ; 1615-9853
    ISSN (online) 1615-9861
    ISSN 1615-9853
    DOI 10.1002/pmic.201100387
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Regulatory considerations for clinical mass spectrometry: multiple reaction monitoring.

    Boja, Emily S / Rodriguez, Henry

    Clinics in laboratory medicine

    2011  Volume 31, Issue 3, Page(s) 443–453

    MeSH term(s) Biomarkers, Tumor/blood ; Biomarkers, Tumor/standards ; Clinical Chemistry Tests/instrumentation ; Clinical Chemistry Tests/methods ; Clinical Chemistry Tests/standards ; Humans ; Mass Spectrometry/methods ; Mass Spectrometry/standards ; Practice Guidelines as Topic ; Proteomics/methods ; Proteomics/standards ; United States ; United States Food and Drug Administration/legislation & jurisprudence ; United States Food and Drug Administration/standards
    Chemical Substances Biomarkers, Tumor
    Language English
    Publishing date 2011-09
    Publishing country United States
    Document type Journal Article
    ZDB-ID 604580-7
    ISSN 1557-9832 ; 0272-2712
    ISSN (online) 1557-9832
    ISSN 0272-2712
    DOI 10.1016/j.cll.2011.07.001
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article: The path to clinical proteomics research: integration of proteomics, genomics, clinical laboratory and regulatory science.

    Boja, Emily S / Rodriguez, Henry

    The Korean journal of laboratory medicine

    2011  Volume 31, Issue 2, Page(s) 61–71

    Abstract: Better biomarkers are urgently needed to cancer detection, diagnosis, and prognosis. While the genomics community is making significant advances in understanding the molecular basis of disease, proteomics will delineate the functional units of a cell, ... ...

    Abstract Better biomarkers are urgently needed to cancer detection, diagnosis, and prognosis. While the genomics community is making significant advances in understanding the molecular basis of disease, proteomics will delineate the functional units of a cell, proteins and their intricate interaction network and signaling pathways for the underlying disease. Great progress has been made to characterize thousands of proteins qualitatively and quantitatively in complex biological systems by utilizing multi-dimensional sample fractionation strategies, mass spectrometry and protein microarrays. Comparative/quantitative analysis of high-quality clinical biospecimen (e.g., tissue and biofluids) of human cancer proteome landscape has the potential to reveal protein/peptide biomarkers responsible for this disease by means of their altered levels of expression, post-translational modifications as well as different forms of protein variants. Despite technological advances in proteomics, major hurdles still exist in every step of the biomarker development pipeline. The National Cancer Institute's Clinical Proteomic Technologies for Cancer initiative (NCI-CPTC) has taken a critical step to close the gap between biomarker discovery and qualification by introducing a pre-clinical "verification" stage in the pipeline, partnering with clinical laboratory organizations to develop and implement common standards, and developing regulatory science documents with the US Food and Drug Administration to educate the proteomics community on analytical evaluation requirements for multiplex assays in order to ensure the safety and effectiveness of these tests for their intended use.
    MeSH term(s) Biomarkers/analysis ; Clinical Laboratory Techniques/standards ; Genomics ; Humans ; Mass Spectrometry/methods ; Mass Spectrometry/standards ; Neoplasms/diagnosis ; Neoplasms/genetics ; Proteomics ; Quality Control ; United States ; United States Food and Drug Administration
    Chemical Substances Biomarkers
    Language English
    Publishing date 2011-04-07
    Publishing country Korea (South)
    Document type Journal Article ; Review
    ZDB-ID 2614035-4
    ISSN 1598-6535
    ISSN 1598-6535
    DOI 10.3343/kjlm.2011.31.2.61
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: Deep Proteomics Using Two Dimensional Data Independent Acquisition Mass Spectrometry.

    Cho, Kyung-Cho / Clark, David J / Schnaubelt, Michael / Teo, Guo Ci / Leprevost, Felipe da Veiga / Bocik, William / Boja, Emily S / Hiltke, Tara / Nesvizhskii, Alexey I / Zhang, Hui

    Analytical chemistry

    2020  Volume 92, Issue 6, Page(s) 4217–4225

    Abstract: Methodologies that facilitate high-throughput proteomic analysis are a key step toward moving proteome investigations into clinical translation. Data independent acquisition (DIA) has potential as a high-throughput analytical method due to the reduced ... ...

    Abstract Methodologies that facilitate high-throughput proteomic analysis are a key step toward moving proteome investigations into clinical translation. Data independent acquisition (DIA) has potential as a high-throughput analytical method due to the reduced time needed for sample analysis, as well as its highly quantitative accuracy. However, a limiting feature of DIA methods is the sensitivity of detection of low abundant proteins and depth of coverage, which other mass spectrometry approaches address by two-dimensional fractionation (2D) to reduce sample complexity during data acquisition. In this study, we developed a 2D-DIA method intended for rapid- and deeper-proteome analysis compared to conventional 1D-DIA analysis. First, we characterized 96 individual fractions obtained from the protein standard, NCI-7, using a data-dependent approach (DDA), identifying a total of 151,366 unique peptides from 11,273 protein groups. We observed that the majority of the proteins can be identified from just a few selected fractions. By performing an optimization analysis, we identified six fractions with high peptide number and uniqueness that can account for 80% of the proteins identified in the entire experiment. These selected fractions were combined into a single sample which was then subjected to DIA (referred to as 2D-DIA) quantitative analysis. Furthermore, improved DIA quantification was achieved using a hybrid spectral library, obtained by combining peptides identified from DDA data with peptides identified directly from the DIA runs with the help of DIA-Umpire. The optimized 2D-DIA method allowed for improved identification and quantification of low abundant proteins compared to conventional unfractionated DIA analysis (1D-DIA). We then applied the 2D-DIA method to profile the proteomes of two breast cancer patient-derived xenograft (PDX) models, quantifying 6,217 and 6,167 unique proteins in basal- and luminal- tumors, respectively. Overall, this study demonstrates the potential of high-throughput quantitative proteomics using a novel 2D-DIA method.
    MeSH term(s) Humans ; Mass Spectrometry ; Peptides/analysis ; Proteins/analysis ; Proteomics
    Chemical Substances Peptides ; Proteins
    Language English
    Publishing date 2020-02-26
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 1508-8
    ISSN 1520-6882 ; 0003-2700
    ISSN (online) 1520-6882
    ISSN 0003-2700
    DOI 10.1021/acs.analchem.9b04418
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article ; Online: Linking cancer genome to proteome: NCI's investment into proteogenomics.

    Rivers, Robert C / Kinsinger, Christopher / Boja, Emily S / Hiltke, Tara / Mesri, Mehdi / Rodriguez, Henry

    Proteomics

    2014  Volume 14, Issue 23-24, Page(s) 2633–2636

    Abstract: Advances in both targeted and unbiased MS-based proteomics are now at a mature stage for comprehensively and reproducibly characterizing a large part of the cancer proteome. These developments combined with the extensive genomic characterization of ... ...

    Abstract Advances in both targeted and unbiased MS-based proteomics are now at a mature stage for comprehensively and reproducibly characterizing a large part of the cancer proteome. These developments combined with the extensive genomic characterization of several cancer types by large-scale initiatives such as the International Cancer Genome Consortium and Cancer Genome Atlas Project have paved the way for proteogenomic analysis of omics datasets and integration methods. The advances serve as the basis for the National Cancer Institute's Clinical Proteomic Tumor Analysis Consortium and this article highlights its current work and future steps in the area of proteogenomics.
    MeSH term(s) Animals ; Genomics/methods ; Humans ; Neoplasms/genetics ; Neoplasms/metabolism ; Proteome/genetics ; Proteome/metabolism ; Proteomics/methods
    Chemical Substances Proteome
    Language English
    Publishing date 2014-12
    Publishing country Germany
    Document type Journal Article
    ZDB-ID 2032093-0
    ISSN 1615-9861 ; 1615-9853
    ISSN (online) 1615-9861
    ISSN 1615-9853
    DOI 10.1002/pmic.201400193
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  8. Article ; Online: Integrated Proteomic and Glycoproteomic Characterization of Human High-Grade Serous Ovarian Carcinoma.

    Hu, Yingwei / Pan, Jianbo / Shah, Punit / Ao, Minghui / Thomas, Stefani N / Liu, Yang / Chen, Lijun / Schnaubelt, Michael / Clark, David J / Rodriguez, Henry / Boja, Emily S / Hiltke, Tara / Kinsinger, Christopher R / Rodland, Karin D / Li, Qing Kay / Qian, Jiang / Zhang, Zhen / Chan, Daniel W / Zhang, Hui

    Cell reports

    2020  Volume 33, Issue 3, Page(s) 108276

    Abstract: Many gene products exhibit great structural heterogeneity because of an array of modifications. These modifications are not directly encoded in the genomic template but often affect the functionality of proteins. Protein glycosylation plays a vital role ... ...

    Abstract Many gene products exhibit great structural heterogeneity because of an array of modifications. These modifications are not directly encoded in the genomic template but often affect the functionality of proteins. Protein glycosylation plays a vital role in proper protein functions. However, the analysis of glycoproteins has been challenging compared with other protein modifications, such as phosphorylation. Here, we perform an integrated proteomic and glycoproteomic analysis of 83 prospectively collected high-grade serous ovarian carcinoma (HGSC) and 23 non-tumor tissues. Integration of the expression data from global proteomics and glycoproteomics reveals tumor-specific glycosylation, uncovers different glycosylation associated with three tumor clusters, and identifies glycosylation enzymes that were correlated with the altered glycosylation. In addition to providing a valuable resource, these results provide insights into the potential roles of glycosylation in the pathogenesis of HGSC, with the possibility of distinguishing pathological outcomes of ovarian tumors from non-tumors, as well as classifying tumor clusters.
    MeSH term(s) Biomarkers, Tumor/metabolism ; Cystadenocarcinoma, Serous/genetics ; Cystadenocarcinoma, Serous/metabolism ; Cystadenocarcinoma, Serous/pathology ; Female ; Glycoproteins/metabolism ; Glycosylation ; Humans ; Ovarian Neoplasms/genetics ; Ovarian Neoplasms/metabolism ; Ovarian Neoplasms/pathology ; Proteomics/methods ; Tissue Banks
    Chemical Substances Biomarkers, Tumor ; Glycoproteins
    Language English
    Publishing date 2020-10-19
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 2649101-1
    ISSN 2211-1247 ; 2211-1247
    ISSN (online) 2211-1247
    ISSN 2211-1247
    DOI 10.1016/j.celrep.2020.108276
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  9. Article ; Online: Analytical validation considerations of multiplex mass-spectrometry-based proteomic platforms for measuring protein biomarkers.

    Boja, Emily S / Fehniger, Thomas E / Baker, Mark S / Marko-Varga, György / Rodriguez, Henry

    Journal of proteome research

    2014  Volume 13, Issue 12, Page(s) 5325–5332

    Abstract: ... our understanding of the molecular basis of disease, with a number of biomarker panels already authorized by the U.S ...

    Abstract Protein biomarker discovery and validation in current omics era are vital for healthcare professionals to improve diagnosis, detect cancers at an early stage, identify the likelihood of cancer recurrence, stratify stages with differential survival outcomes, and monitor therapeutic responses. The success of such biomarkers would have a huge impact on how we improve the diagnosis and treatment of patients and alleviate the financial burden of healthcare systems. In the past, the genomics community (mostly through large-scale, deep genomic sequencing technologies) has been steadily improving our understanding of the molecular basis of disease, with a number of biomarker panels already authorized by the U.S. Food and Drug Administration (FDA) for clinical use (e.g., MammaPrint, two recently cleared devices using next-generation sequencing platforms to detect DNA changes in the cystic fibrosis transmembrane conductance regulator (CFTR) gene). Clinical proteomics, on the other hand, albeit its ability to delineate the functional units of a cell, more likely driving the phenotypic differences of a disease (i.e., proteins and protein-protein interaction networks and signaling pathways underlying the disease), "staggers" to make a significant impact with only an average ∼ 1.5 protein biomarkers per year approved by the FDA over the past 15-20 years. This statistic itself raises the concern that major roadblocks have been impeding an efficient transition of protein marker candidates in biomarker development despite major technological advances in proteomics in recent years.
    MeSH term(s) Biomarkers, Tumor/analysis ; Biomarkers, Tumor/metabolism ; Biomedical Research/methods ; Biomedical Research/standards ; Biomedical Research/trends ; Computational Biology/methods ; Computational Biology/standards ; Computational Biology/trends ; Humans ; Mass Spectrometry/instrumentation ; Mass Spectrometry/methods ; Neoplasms/diagnosis ; Neoplasms/metabolism ; Proteome/analysis ; Proteome/metabolism ; Proteomics/methods ; Proteomics/standards ; Proteomics/trends ; Reproducibility of Results
    Chemical Substances Biomarkers, Tumor ; Proteome
    Language English
    Publishing date 2014-11-18
    Publishing country United States
    Document type Journal Article ; Review
    ZDB-ID 2078618-9
    ISSN 1535-3907 ; 1535-3893
    ISSN (online) 1535-3907
    ISSN 1535-3893
    DOI 10.1021/pr500753r
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  10. Article ; Online: Four areas of engagement requiring strengthening in modern proteomics today.

    Fehniger, Thomas E / Boja, Emily S / Rodriguez, Henry / Baker, Mark S / Marko-Varga, György

    Journal of proteome research

    2014  Volume 13, Issue 12, Page(s) 5310–5318

    Abstract: ... response to therapy into the approval processes of regulatory agencies (e.g., U.S ...

    Abstract The global human proteomics community in 2014 is fully engaged in projects that aim to create a better understanding of human biology and its complexities and to provide products from this new knowledge that will in some way benefit humanity. Human proteomics, like any other scientific enterprise, needs to identify areas of direction and development, both for the near future in completing current research projects and into the long-term for the engagement with even more complex challenges. In this Editorial we highlight and discuss four important areas that we collectively believe require attention and demand a collective response going forward. These four areas are: (1) Provide high-quality standardized, sensitive, specific, quantitative, and readily accessible protein, peptide, or other biomarkers of health, disease, response to therapy into the approval processes of regulatory agencies (e.g., U.S. Food and Drug Administration; FDA), and obtaining approval from the relevant agencies for their use in a clinical or other testing settings. (2) Implement standard processes for collecting, processing, and storing human clinical samples in biorepositories and enforcement of measures to ensure subject integrity including informed consent for the downstream use of samples and in registrations of subject identities within study databases. (3) Test and validate mass spectrometry technology platforms that hold much promise for creating opportunities for obtaining new important knowledge at levels of detection previously not achievable. (4) Organize clinical discovery operations and activities in an intuitive manner to meet the challenges of increased interests in the science we provide and diminishing levels of centrally financed resource and infrastructure support.
    MeSH term(s) Biomarkers/analysis ; Biomarkers/metabolism ; Biomedical Research/methods ; Biomedical Research/standards ; Biomedical Research/trends ; Humans ; Mass Spectrometry/methods ; Proteome/analysis ; Proteome/metabolism ; Proteomics/methods ; Proteomics/standards ; Proteomics/trends ; Reference Standards ; Reproducibility of Results
    Chemical Substances Biomarkers ; Proteome
    Language English
    Publishing date 2014-12-05
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2078618-9
    ISSN 1535-3907 ; 1535-3893
    ISSN (online) 1535-3907
    ISSN 1535-3893
    DOI 10.1021/pr500472d
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