Principal Investigator

Richard R.
Drake
Awardee Organization

Medical University Of South Carolina
United States

Fiscal Year
2024
Activity Code
R33
Early Stage Investigator Grants (ESI)
Not Applicable
Project End Date

Targeted Isolation and Identification of Sialylated Glycoproteins in Cancer Tissues, Cells and Biofluids

Glycoproteins account for the majority of the proteins located at the cell surface and in the extracellular environment. During carcinogenesis the normal function of these glycoproteins as binding ligands for cell-cell adhesion, extracellular matrix molecules, signaling receptors, and immune cells becomes altered. Many of the steps in metastatic progression involve re-organization and degradation of the glycoproteins in the extracellular matrix. Hence, glycan and glycoprotein antigens still comprise the majority of FDA approved diagnostic cancer biomarkers. Our group has recently developed multiple N-linked glycan imaging mass spectrometry workflows to characterize the glycomes in clinically relevant tumor tissues, biofluids and cells, as well as individually targeted glycoproteins by immunocapture. In this proposal, application of a new click chemistry reagent specific for targeting of α2,3 sialic acid antigens, like sialyl Lewis A/CA19-9, sialyl Lewis X and sTRA, will be used to improve isolation and sequencing of tumor-associated glycopeptides from FFPE tissues, biofluids and cells. Standard proteases and new ECM-targeted proteases will be used in the analyses. To accomplish this, three Specific Aims are proposed: 1. Isolate, identify and co-localize sialic acid glycoproteins from clinical FFPE cancer tissues; 2. Isolate and identify sialic acid glycoproteins in plasma cancer samples; 3. Isolate and identify sialic acid glycoproteins in cancer cell lines with high levels of α2,3 sialic antigens. The carbohydrate-targeted click capture reagents developed can be applied directly to any FFPE tissue, cell or biofluid of interest using simple chemical reactions. The combination of tools developed herein will facilitate the identification of specific glycopeptide and glycoprotein species in relevant cancer samples which can be applied across the spectrum of cancer prevention, progression, prognosis, and biomarker applications.

Publications

  • Wallace EN, West CA, McDowell CT, Lu X, Bruner E, Mehta AS, Aoki-Kinoshita KF, Angel PM, Drake RR. An N-glycome tissue atlas of 15 human normal and cancer tissue types determined by MALDI-imaging mass spectrometry. Scientific reports. 2024 Jan 4;14(1):489. PMID: 38177192
  • Lu X, McDowell CT, Blaschke CRK, Liu L, Grimsley G, Wisniewski L, Gao C, Mehta AS, Haab BB, Angel PM, Drake RR. Bioorthogonal Chemical Labeling Probes Targeting Sialic Acid Isomers for N-Glycan MALDI Imaging Mass Spectrometry of Tissues, Cells, and Biofluids. Analytical chemistry. 2023 May 16;95(19):7475-7486. Epub 2023 May 1. PMID: 37126482
  • Young LEA, Nietert PJ, Stubler R, Kittrell CG, Grimsley G, Lewin DN, Mehta AS, Hajar C, Wang K, O'Quinn EC, Angel PM, Wallace K, Drake RR. Utilizing multimodal mass spectrometry imaging for profiling immune cell composition and N-glycosylation across colorectal carcinoma disease progression. Frontiers in pharmacology. 2024 Jan 11;14:1337319. doi: 10.3389/fphar.2023.1337319. eCollection 2023. PMID: 38273829