Program Official

Principal Investigator

Awardee Organization

Georgetown University
United States

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

O-glycoproteins in the progression of liver disease

We propose to apply targeted mass spectrometric methods to the analysis of specific O-glycoforms of proteins in the progression of liver disease to hepatocellular carcinoma (HCC). We have identified novel sialylated Oglycoform of liver secreted proteins significantly increased at progressive stages of liver disease. We propose to determine structure of the glycoforms and to evaluate extent of their presence on liver secreted glycoproteins. We will study association of the O-glycoforms with progressing stages of liver pathology by their direct quantification at specific sites of protein attachment in serum of patients. The O-glycoforms will be analyzed in combination with other markers identified in previous studies. We combine current concepts of disease mechanisms and advanced technologies to design improved serologic assays expected to complement biopsy and other non-invasive procedures in the assessment of liver disease. Non-invasive monitoring of liver disease progression to HCC is expected to influence optimal selection of therapeutic approaches and improve disease outcomes.


  • Yuan W, Wei R, Goldman R, Sanda M. Optimized Fragmentation for Quantitative Analysis of Fucosylated N-Glycoproteins by LC-MS-MRM. Analytical chemistry. 2019 Jul 16;91(14):9206-9212. Epub 2019 Jul 3. PMID: 31268672
  • McIntosh AT, Wei R, Ahn J, Aouizerat BE, Kassaye SG, Augenbraun MH, Price JC, French AL, Gange SJ, Anastos KM, Goldman R. A genomic variant of ALPK2 is associated with increased liver fibrosis risk in HIV/HCV coinfected women. PloS one. 2021 Mar 11;16(3):e0247277. doi: 10.1371/journal.pone.0247277. eCollection 2021. PMID: 33705408
  • Panigrahi A, Benicky J, Aljuhani R, Mukherjee P, Nováková Z, Bařinka C, Goldman R. Galectin-3-binding protein inhibits extracellular heparan 6-O-endosulfatse Sulf-2. bioRxiv : the preprint server for biology. 2023 Dec 21. PMID: 38187586
  • Mukherjee P, Zhou X, Benicky J, Panigrahi A, Aljuhani R, Liu J, Ailles L, Pomin VH, Wang Z, Goldman R. Heparan-6-O-Endosulfatase 2 Promotes Invasiveness of Head and Neck Squamous Carcinoma Cell Lines in Co-Cultures with Cancer-Associated Fibroblasts. Cancers. 2023 Oct 27;15. (21). PMID: 37958342
  • Benicky J, Sanda M, Brnakova Kennedy Z, Grant OC, Woods RJ, Zwart A, Goldman R. PD-L1 Glycosylation and Its Impact on Binding to Clinical Antibodies. Journal of proteome research. 2021 Jan 1;20(1):485-497. Epub 2020 Oct 19. PMID: 33073996
  • Benicky J, Sanda M, Brnakova Kennedy Z, Goldman R. N-Glycosylation is required for secretion of the precursor to brain-derived neurotrophic factor (proBDNF) carrying sulfated LacdiNAc structures. The Journal of biological chemistry. 2019 Nov 8;294(45):16816-16830. Epub 2019 Sep 26. PMID: 31558607
  • Sanda M, Yang Q, Zong G, Chen H, Zheng Z, Dhani H, Khan K, Kroemer A, Wang LX, Goldman R. LC-MS/MS-PRM Quantification of IgG Glycoforms Using Stable Isotope Labeled IgG1 Fc Glycopeptide Standard. Journal of proteome research. 2023 Apr 7;22(4):1138-1147. Epub 2023 Feb 10. PMID: 36763792
  • Kozlik P, Molnarova K, Jecmen T, Krizek T, Goldman R. Glycan-specific precipitation of glycopeptides in high organic content sample solvents used in HILIC. Journal of chromatography. B, Analytical technologies in the biomedical and life sciences. 2020 Aug 1;1150:122196. Epub 2020 May 28. PMID: 32485649
  • York WS, Mazumder R, Ranzinger R, Edwards N, Kahsay R, Aoki-Kinoshita KF, Campbell MP, Cummings RD, Feizi T, Martin M, Natale DA, Packer NH, Woods RJ, Agarwal G, Arpinar S, Bhat S, Blake J, Castro LJG, Fochtman B, Gildersleeve J, Goldman R, Holmes X, Jain V, Kulkarni S, Mahadik R, Mehta A, Mousavi R, Nakarakommula S, Navelkar R, Pattabiraman N, Pierce MJ, Ross K, Vasudev P, Vora J, Williamson T, Zhang W. GlyGen: Computational and Informatics Resources for Glycoscience. Glycobiology. 2020 Jan 28;30(2):72-73. PMID: 31616925
  • Panigrahi A, Benicky J, Wei R, Ahn J, Goldman R, Sanda M. A Rapid LC-MS/MS-PRM Assay for Serologic Quantification of Sialylated O-HPX Glycoforms in Patients with Liver Fibrosis. Molecules (Basel, Switzerland). 2022 Mar 29;27. (7). PMID: 35408612
  • Yang Y, Ahn J, Edwards NJ, Benicky J, Rozeboom AM, Davidson B, Karamboulas C, Nixon KCJ, Ailles L, Goldman R. Extracellular Heparan 6-O-Endosulfatases SULF1 and SULF2 in Head and Neck Squamous Cell Carcinoma and Other Malignancies. Cancers. 2022 Nov 11;14. (22). PMID: 36428645
  • Sanda M, Benicky J, Goldman R. Low Collision Energy Fragmentation in Structure-Specific Glycoproteomics Analysis. Analytical chemistry. 2020 Jun 16;92(12):8262-8267. Epub 2020 Jun 4. PMID: 32441515
  • Kawahara R, Chernykh A, Alagesan K, Bern M, Cao W, Chalkley RJ, Cheng K, Choo MS, Edwards N, Goldman R, Hoffmann M, Hu Y, Huang Y, Kim JY, Kletter D, Liquet B, Liu M, Mechref Y, Meng B, Neelamegham S, Nguyen-Khuong T, Nilsson J, Pap A, Park GW, Parker BL, Pegg CL, Penninger JM, Phung TK, Pioch M, Rapp E, Sakalli E, Sanda M, Schulz BL, Scott NE, Sofronov G, Stadlmann J, Vakhrushev SY, Woo CM, Wu HY, Yang P, Ying W, Zhang H, Zhang Y, Zhao J, Zaia J, Haslam SM, Palmisano G, Yoo JS, Larson G, Khoo KH, Medzihradszky KF, Kolarich D, Packer NH, Thaysen-Andersen M. Community evaluation of glycoproteomics informatics solutions reveals high-performance search strategies for serum glycopeptide analysis. Nature methods. 2021 Nov;18(11):1304-1316. Epub 2021 Nov 1. PMID: 34725484
  • Benicky J, Sanda M, Panigrahi A, Liu J, Wang Z, Pagadala V, Su G, Goldman R. A 6-O-endosulfatase activity assay based on synthetic heparan sulfate oligomers. Glycobiology. 2023 Jun 3;33(5):384-395. PMID: 37052463
  • Yang Y, Ahn J, Raghunathan R, Kallakury BV, Davidson B, Kennedy ZB, Zaia J, Goldman R. Expression of the Extracellular Sulfatase SULF2 Affects Survival of Head and Neck Squamous Cell Carcinoma Patients. Frontiers in oncology. 2021 Jan 8;10:582827. doi: 10.3389/fonc.2020.582827. eCollection 2020. PMID: 33585200
  • Sanda M, Ahn J, Kozlik P, Goldman R. Analysis of site and structure specific core fucosylation in liver cirrhosis using exoglycosidase-assisted data-independent LC-MS/MS. Scientific reports. 2021 Dec 2;11(1):23273. PMID: 34857845
  • Panigrahi A, Zhang L, Benicky J, Sanda M, Ahn J, Goldman R. A multiplexed microflow LC-MS/MS-PRM assay for serologic quantification of IgG N- and HPX O- glycoforms in liver fibrosis. Scientific reports. 2023 Jan 12;13(1):606. PMID: 36635317
  • Sanda M, Morrison L, Goldman R. N- and O-Glycosylation of the SARS-CoV-2 Spike Protein. Analytical chemistry. 2021 Feb 2;93(4):2003-2009. Epub 2021 Jan 6. PMID: 33406838