Two-Dimensional N-Glycan Distribution Mapping of Hepatocellular Carcinoma Tissues by MALDI-Imaging Mass Spectrometry.
A new mass spectrometry imaging approach to simultaneously map the two-dimensional distribution of N-glycans in tissues has been recently developed. The method uses Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) to spatially profile the location and distribution of multiple N-linked glycan species released by peptide N-glycosidase F in frozen or formalin-fixed tissues. Multiple formalin-fixed human hepatocellular carcinoma tissues were evaluated with this method, resulting in a panel of over 30 N-glycans detected. An ethylation reaction of extracted N-glycans released from adjacent slides was done to stabilize sialic acid containing glycans, and these structures were compared to N-glycans detected directly from tissue profiling. In addition, the distribution of singly fucosylated N-glycans detected in tumor tissue microarray cores were compared to the histochemistry staining pattern of a core fucose binding lectin. As this MALDI-IMS workflow has the potential to be applied to any formalin-fixed tissue block or tissue microarray, the advantages and limitations of the technique in context with other glycomic methods are also summarized.
Department of Cell and Molecular Pharmacology and Experimental Therapeutics and MUSC Proteomics Center, Medical University of South Carolina, 173 Ashley Avenue, Charleston, SC 29425, USA. ThomasWesley.Powers@pfizer.com.
Thomas W
TW
Powers
Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden 2333ZA, The Netherlands. s.holst@lumc.nl.
Stephanie
S
Holst
Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden 2333ZA, The Netherlands. m.wuhrer@lumc.nl.|Division of BioAnalytical Chemistry, VU University, Amsterdam 1081HV, The Netherlands. m.wuhrer@lumc.nl.|Department of Molecular Cell Biology and Immunology, VU University Medical Center, Amsterdam 1007MB, The Netherlands. m.wuhrer@lumc.nl.
Manfred
M
Wuhrer
Department of Microbiology and Immunology, College of Medicine, Drexel University, 2900 W. Queen Lane, Philadelphia, PA 19129, USA. anand.mehta@drexelmed.edu.
Anand S
AS
Mehta
Department of Cell and Molecular Pharmacology and Experimental Therapeutics and MUSC Proteomics Center, Medical University of South Carolina, 173 Ashley Avenue, Charleston, SC 29425, USA. draker@musc.edu.
Richard R
RR
Drake
D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D005557:Formaldehyde; D006031:Glycosylation; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism*; D011134:Polysaccharides / Q000032:analysis / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D011134:Polysaccharides; D005557:Formaldehyde
MALDI imaging mass spectrometry; N-linked glycosylation; formalin-fixed paraffin-embedded tissue; glycoprotein; hepatocellular carcinoma