Ultra-performance ion-pairing liquid chromatography with on-line electrospray ion trap mass spectrometry for heparin disaccharide analysis

Yang, Bo; Weyers, Amanda; Baik, Jong Youn; Sterner, Eric; Sharfstein, Susan T.; Mousa, Shaker A.; Zhang, Fuming; Dordick, Jonathan S.; Linhardt, Robert J.

doi:10.1016/j.ab.2011.04.003

Cited by 71 publications

(78 citation statements)

References 51 publications

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“…GAG chains were purified and analyzed from tumorsphere cultures following the protocol reported by Yang B. et al (2011) (37) with minor modifications (see Supplemental Materials and Methods). The HS disaccharides were released from the GAG chains with heparitinase I, II, III (a kind gift from Dr. Kuberan Balagurunathan) in heparitinase buffer (3.3 mM calcium acetate, 40 mM amonium acetate, 0.1mg/ml BSA, pH 7.0) overnight followed by enzyme inactivation at 100°C for 5 min.…”

Section: Methodsmentioning

confidence: 99%

Heparan Sulfate Glycosaminoglycans in Glioblastoma Promote Tumor Invasion

Tran

Wade

McKinney

et al. 2017

Molecular Cancer Research

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Glioblastoma (GBM) is the most common primary malignant brain tumor of adults and confers a poor prognosis due, in part, to diffuse invasion of tumor cells. Heparan sulfate (HS) glycosaminoglycans, present on the cell surface and in the extracellular matrix, regulate cell signaling pathways and cell-microenvironment interactions. In GBM, the expression of HS glycosaminoglycans and the enzymes that regulate their function are altered but the actual HS content and structure are unknown. However, inhibition of HS glycosaminoglycan function is emerging as a promising therapeutic strategy for some cancers. In this study, we use liquid chromatography-mass spectrometry (LC/MS) analysis to demonstrate differences in HS disaccharide content and structure across four patient-derived tumorsphere lines (GBM1, 5, 6, 43) and between two murine tumorsphere lines derived from murine GBM with enrichment of mesenchymal and proneural gene expression (mMES and mPN, respectively) markers. In GBM, the heterogeneous HS content and structure across patient-derived tumorsphere lines suggested diverse functions in the GBM tumor microenvironment (TME). In GBM5 and mPN, elevated expression of sulfatase 2 (SULF2), an extracellular enzyme that alters ligand binding to HS, was associated with low trisulfated HS disaccharides, a substrate of SULF2. In contrast, other primary tumorsphere lines had elevated expression of the HS-modifying enzyme heparanase (HPSE). Using gene editing strategies to inhibit HPSE a role for HPSE in promoting tumor cell adhesion and invasion was identified. These studies characterize the heterogeneity in HS glycosaminoglycan content and structure across GBM and reveal their role in tumor cell invasion.

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Section: Methodsmentioning

confidence: 99%

Heparan Sulfate Glycosaminoglycans in Glioblastoma Promote Tumor Invasion

Tran

Wade

McKinney

et al. 2017

Molecular Cancer Research

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“…Glycosaminoglycan Isolation and Purification-Isolation of glycosaminoglycans from media samples has been described previously (20,21). The plasma samples were lyophilized and defatted and then individually subjected to proteolysis at 55°C with 10% (w/v) of actinase E (20 mg/ml in HPLC grade water, Kaken Biochemicals, Tokyo, Japan) for 20 h. After proteolysis, particulates were removed from the resulting solutions by passing each through a 0.22-m membrane syringe filter.…”

Section: Methodsmentioning

confidence: 99%

The Circulating Glycosaminoglycan Signature of Respiratory Failure in Critically Ill Adults

Schmidt

et al. 2014

Journal of Biological Chemistry

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134

137

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Background: Endothelial glycocalyx degradation contributes to the pathogenesis of critical illness. Results: Mechanically ventilated subjects exhibited plasma glycocalyx breakdown signatures (glycosaminoglycan fragments) characteristic of direct versus indirect etiologies of respiratory failure. Conclusion: Circulating glycosaminoglycans provide insight into respiratory failure pathophysiology. Significance: This is the first study to characterize circulating glycosaminoglycans during critical illness, offering insight into the mechanisms underlying respiratory failure.

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“…The electrospray interface was set in negative ionization mode with the skimmer potential -40.0 V, capillary exit -40.0 V, and a source of temperature of 350°C to obtain maximum abundance of the ions in a full-scan spectra (350-1500 Da, 10 full scans/s). Nitrogen was used as a drying gas (8 L/min) and a nebulizing gas (40 psi) (Yang et al, 2011).…”

Section: Analysis Of Isolated Gags Using Pagementioning

confidence: 99%

A Structural Analysis of Glycosaminoglycans from Lethal and Nonlethal Breast Cancer Tissues: Toward a Novel Class of Theragnostics for Personalized Medicine in Oncology?

Weyers

Yang

Yoon

et al. 2012

OMICS: A Journal of Integrative Biology

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Cancer is one of the leading noncommunicable diseases that vastly impacts both developed and developing countries. Truly innovative diagnostics that inform disease susceptibility, prognosis, and/or response to treatment (theragnostics) are seriously needed for global public health and personalized medicine for patients with cancer. This study examined the structure and content of glycosaminoglycans (GAGs) in lethal and nonlethal breast cancer tissues from six patients. The glycosaminoglycan content isolated from tissue containing lethal cancer tumors was approximately twice that of other tissues. Molecular weight analysis showed that glycosaminoglycans from cancerous tissue had a longer weight average chain length by an average of five disaccharide units, an increase of approximately 15%. Dissacharide analysis found differences in sulfation patterns between cancerous and normal tissues, as well as sulfation differences in GAG chains isolated from patients with lethal and nonlethal cancer. Specifically, cancerous tissue showed an increase in sulfation at the ''6S'' position of CS chains and an increase in the levels of the HS disaccharide NSCS. Patients with lethal cancer showed a decrease in HS sulfation, with lower levels of ''6S'' and higher levels of the unsulfated ''0S'' disaccharide. Although these findings come from a limited sample size, they indicate that structural changes in GAGs exist between cancerous and noncancerous tissues and between tissues from patients with highly metastatic cancer and cancer that was successfully treated by chemotherapy. Based on these findings, we hypothesize that (1) there are putative changes in the body's construction of GAGs as tissue becomes cancerous; (2) there may be innate structural person-to-person variations in GAG composition that facilitate the metastasis of tumors in some patients when they develop cancer.

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Ultra-performance ion-pairing liquid chromatography with on-line electrospray ion trap mass spectrometry for heparin disaccharide analysis

Cited by 71 publications

References 51 publications

Heparan Sulfate Glycosaminoglycans in Glioblastoma Promote Tumor Invasion

Heparan Sulfate Glycosaminoglycans in Glioblastoma Promote Tumor Invasion

The Circulating Glycosaminoglycan Signature of Respiratory Failure in Critically Ill Adults

A Structural Analysis of Glycosaminoglycans from Lethal and Nonlethal Breast Cancer Tissues: Toward a Novel Class of Theragnostics for Personalized Medicine in Oncology?

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