Dataset Information

Identification and relative quantification of hFSH glycoforms in women sera by PRM MS based approach.

ABSTRACT: This study presents an alternative preliminary strategy to identify and quantify serum hFSH glycoforms based on immunopurification assay and mass spectrometry (MS) using parallel reaction monitoring (PRM) analysis. In this study, we provide MS-PRM data acquisition method for hFSH glycopeptides identification with high specificity and their quantification by extracting the chromatographic traces of selected fragments of glycopeptides. Once set up in all its features, the proposed method could be transferred to the clinic to improve the fertility treatments and follow-ups in men and women.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood Serum

SUBMITTER: Chiara Melchiorre

LAB HEAD: Ida Chiara Guerrera

PROVIDER: PXD025585 | Pride | 2021-06-15

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	200129_fsh_Chymotry_sol_ChME_rhs_gonalf_zt_Q31657.raw	Raw
	200129_fsh_Chymotry_sol_ChME_rhs_gonalf_zt_Q31657.xml	Xml
	200228_ChME_IP_ChME_FSH_serum_zt_Q31824.raw	Raw
	200228_ChME_IP_ChME_FSH_serum_zt_Q31824.xml	Xml
	200228_ChME_IP_FSH_serum_zt_PRM_Q31826.raw	Raw

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Publications

Identification and Relative Quantification of hFSH Glycoforms in Women's Sera via MS-PRM-Based Approach.

Melchiorre Chiara C Chhuon Cerina C Jung Vincent V Lipecka Joanna J Di Rella Francesca F Conforti Alessandro A Amoresano Angela A Carpentieri Andrea A Guerrera Ida Chiara IC

Pharmaceutics 20210527 6

Follicle-stimulating hormone (FSH) is a glycohormone synthesized by adenohypophysis, and it stimulates ovulation in women and spermatogenesis in men by binding to its receptor (FSHR). FSHR is involved in several mechanisms to transduce intracellular signals in response to the FSH stimulus. Exogenous FSH is currently used in the clinic for ovarian hyperstimulation during in vitro fertilization in women, and for treatment of infertility caused by gonadotropin deficiency in men. The glycosylation o ...[more]

PMID: 34071747

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Project description:The most common congenital disorder of glycosylation (CDG) is caused by pathogenic variants in PMM2 (PMM2-CDG) that reduce phosphomannomutase activity and impair N-glycan synthesis. Patients present early in life with multi-system disabilities. Traditional diagnosis relies on measuring carbohydrate-deficient transferrin (CDT) and confirmation by genetic testing. However, tests for CDT can be normal in some patients, or normalize with age. Because patients also show subtle alterations in serum glycan structures, we considered that site-specific glycosylation changes might be diagnostic. We analyzed serum from 35 individuals with PMM2-CDG for discovery and validation of glycoproteomic changes. We carried out mass spectrometry-based discovery studies to profile the glycoproteome in an initial set of well-characterized affected individuals and matched controls. Next, we developed a streamlined method for analyzing additional affected individuals. Finally, targeted mass spectrometry was used to validate selected N-glycopeptides in an independent set of samples in a blinded fashion. Of the 3,342 N-glycopeptides derived from 284 glycoproteins identified in discovery experiments, individuals with PMM2-CDG exhibited a general decrease in complex-type N-glycans and an increase in truncated, mannose-rich and hybrid species. We identified a glycopeptide from complement C4 carrying the glycan Man5GlcNAc2, which was not detected in controls. Notably, this glycopeptide was also detected in five affected individuals with normal CDT, including one individual after liver transplant, two individuals with a known genetic variant associated with mild disease and two individuals with well characterized variants, indicating that this glycopeptide is more sensitive than CDT. Finally, this marker was also detected by targeted analysis in two individuals with variants of uncertain significance (VUS) in PMM2 gene. Overall, we believe that C4-derived Man5GlcNAc2 glycopeptide could serve as a novel next-generation biomarker for more accurate diagnosis and tracking progress of patients in therapeutic trials.