Hypoglycosylated hFSH has greater bioactivity than fully glycosylated recombinant hFSH in human granulosa cells

Chao Jiang, Xiaoying Hou, Cheng Wang, Jeffrey V. May, Viktor Y. Butnev, George R. Bousfield, John S Davis

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Context: Previous studies suggest that aging in women is associated with a reduction in hypoglycosylated forms of FSH. Objective: Experiments were performed to determine whether glycosylation of the FSHβ subunit modulates the biological activity of FSH in human granulosa cells. Design and Setting: Recombinant human FSH (hFSH) derived from GH<inf>3</inf> pituitary cells was purified into fractions containing hypoglycosylated hFSH<sup>21/18</sup> and fully glycosylated hFSH<sup>24</sup>. The response to FSH glycoforms was evaluated using the well-characterized, FSH-responsive human granulosa cell line, KGN at an academic medical center. Interventions: Granulosa cells were treated with increasing concentrations of fully- or hypoglycosylated FSH glycoforms for periods up to 48 hours. Main Outcome Measure(s): The main outcomes were indices of cAMP-dependent cell signaling and estrogen and progesterone synthesis. Results: We observed that hypoglycosylated FSH<sup>21/18</sup> was significantly more effective than fully glycosylated FSH<sup>24</sup> at stimulating cAMP accumulation, protein kinase A (PKA) activity, and cAMP response element binding protein (CREB) (S133) phosphorylation. FSH<sup>21/18</sup> was also much more effective than hFSH<sup>24</sup> on the stimulation CREB-response element-mediated transcription, expression of aromatase and STAR proteins, and synthesis of estrogen and progesterone. Adenoviralmediated expression of the endogenous inhibitor of PKA, inhibited FSH<sup>21/18</sup> - and FSH<sup>24</sup> -stimulated CREB phosphorylation, and steroidogenesis. Conclusions: Hypoglycosylated FSH<sup>21/18</sup> has greater bioactivity than fully glycosylated hFSH<sup>24</sup>, suggesting that age-dependent decreases in hypoglycosylated hFSH contribute to reduced ovarian responsiveness. Hypoglycosylated FSH may be useful in follicle stimulation protocols for older patients using assisted reproduction technologies.

Original languageEnglish (US)
Pages (from-to)E852-E860
JournalJournal of Clinical Endocrinology and Metabolism
Volume100
Issue number6
DOIs
StatePublished - Jun 1 2015

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Human Follicle Stimulating Hormone
Cyclic AMP Response Element-Binding Protein
Granulosa Cells
Bioactivity
Phosphorylation
Cyclic AMP-Dependent Protein Kinases
Progesterone
Estrogens
Cell signaling
Glycosylation
Aromatase
Response Elements
Transcription
Aging of materials
Cells
Reproduction
Outcome Assessment (Health Care)
Technology
Cell Line
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry
  • Endocrinology
  • Biochemistry, medical
  • Endocrinology, Diabetes and Metabolism

Cite this

Hypoglycosylated hFSH has greater bioactivity than fully glycosylated recombinant hFSH in human granulosa cells. / Jiang, Chao; Hou, Xiaoying; Wang, Cheng; May, Jeffrey V.; Butnev, Viktor Y.; Bousfield, George R.; Davis, John S.

In: Journal of Clinical Endocrinology and Metabolism, Vol. 100, No. 6, 01.06.2015, p. E852-E860.

Research output: Contribution to journalArticle

Jiang, Chao ; Hou, Xiaoying ; Wang, Cheng ; May, Jeffrey V. ; Butnev, Viktor Y. ; Bousfield, George R. ; Davis, John S. / Hypoglycosylated hFSH has greater bioactivity than fully glycosylated recombinant hFSH in human granulosa cells. In: Journal of Clinical Endocrinology and Metabolism. 2015 ; Vol. 100, No. 6. pp. E852-E860.
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abstract = "Context: Previous studies suggest that aging in women is associated with a reduction in hypoglycosylated forms of FSH. Objective: Experiments were performed to determine whether glycosylation of the FSHβ subunit modulates the biological activity of FSH in human granulosa cells. Design and Setting: Recombinant human FSH (hFSH) derived from GH3 pituitary cells was purified into fractions containing hypoglycosylated hFSH21/18 and fully glycosylated hFSH24. The response to FSH glycoforms was evaluated using the well-characterized, FSH-responsive human granulosa cell line, KGN at an academic medical center. Interventions: Granulosa cells were treated with increasing concentrations of fully- or hypoglycosylated FSH glycoforms for periods up to 48 hours. Main Outcome Measure(s): The main outcomes were indices of cAMP-dependent cell signaling and estrogen and progesterone synthesis. Results: We observed that hypoglycosylated FSH21/18 was significantly more effective than fully glycosylated FSH24 at stimulating cAMP accumulation, protein kinase A (PKA) activity, and cAMP response element binding protein (CREB) (S133) phosphorylation. FSH21/18 was also much more effective than hFSH24 on the stimulation CREB-response element-mediated transcription, expression of aromatase and STAR proteins, and synthesis of estrogen and progesterone. Adenoviralmediated expression of the endogenous inhibitor of PKA, inhibited FSH21/18 - and FSH24 -stimulated CREB phosphorylation, and steroidogenesis. Conclusions: Hypoglycosylated FSH21/18 has greater bioactivity than fully glycosylated hFSH24, suggesting that age-dependent decreases in hypoglycosylated hFSH contribute to reduced ovarian responsiveness. Hypoglycosylated FSH may be useful in follicle stimulation protocols for older patients using assisted reproduction technologies.",
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T1 - Hypoglycosylated hFSH has greater bioactivity than fully glycosylated recombinant hFSH in human granulosa cells

AU - Jiang, Chao

AU - Hou, Xiaoying

AU - Wang, Cheng

AU - May, Jeffrey V.

AU - Butnev, Viktor Y.

AU - Bousfield, George R.

AU - Davis, John S

PY - 2015/6/1

Y1 - 2015/6/1

N2 - Context: Previous studies suggest that aging in women is associated with a reduction in hypoglycosylated forms of FSH. Objective: Experiments were performed to determine whether glycosylation of the FSHβ subunit modulates the biological activity of FSH in human granulosa cells. Design and Setting: Recombinant human FSH (hFSH) derived from GH3 pituitary cells was purified into fractions containing hypoglycosylated hFSH21/18 and fully glycosylated hFSH24. The response to FSH glycoforms was evaluated using the well-characterized, FSH-responsive human granulosa cell line, KGN at an academic medical center. Interventions: Granulosa cells were treated with increasing concentrations of fully- or hypoglycosylated FSH glycoforms for periods up to 48 hours. Main Outcome Measure(s): The main outcomes were indices of cAMP-dependent cell signaling and estrogen and progesterone synthesis. Results: We observed that hypoglycosylated FSH21/18 was significantly more effective than fully glycosylated FSH24 at stimulating cAMP accumulation, protein kinase A (PKA) activity, and cAMP response element binding protein (CREB) (S133) phosphorylation. FSH21/18 was also much more effective than hFSH24 on the stimulation CREB-response element-mediated transcription, expression of aromatase and STAR proteins, and synthesis of estrogen and progesterone. Adenoviralmediated expression of the endogenous inhibitor of PKA, inhibited FSH21/18 - and FSH24 -stimulated CREB phosphorylation, and steroidogenesis. Conclusions: Hypoglycosylated FSH21/18 has greater bioactivity than fully glycosylated hFSH24, suggesting that age-dependent decreases in hypoglycosylated hFSH contribute to reduced ovarian responsiveness. Hypoglycosylated FSH may be useful in follicle stimulation protocols for older patients using assisted reproduction technologies.

AB - Context: Previous studies suggest that aging in women is associated with a reduction in hypoglycosylated forms of FSH. Objective: Experiments were performed to determine whether glycosylation of the FSHβ subunit modulates the biological activity of FSH in human granulosa cells. Design and Setting: Recombinant human FSH (hFSH) derived from GH3 pituitary cells was purified into fractions containing hypoglycosylated hFSH21/18 and fully glycosylated hFSH24. The response to FSH glycoforms was evaluated using the well-characterized, FSH-responsive human granulosa cell line, KGN at an academic medical center. Interventions: Granulosa cells were treated with increasing concentrations of fully- or hypoglycosylated FSH glycoforms for periods up to 48 hours. Main Outcome Measure(s): The main outcomes were indices of cAMP-dependent cell signaling and estrogen and progesterone synthesis. Results: We observed that hypoglycosylated FSH21/18 was significantly more effective than fully glycosylated FSH24 at stimulating cAMP accumulation, protein kinase A (PKA) activity, and cAMP response element binding protein (CREB) (S133) phosphorylation. FSH21/18 was also much more effective than hFSH24 on the stimulation CREB-response element-mediated transcription, expression of aromatase and STAR proteins, and synthesis of estrogen and progesterone. Adenoviralmediated expression of the endogenous inhibitor of PKA, inhibited FSH21/18 - and FSH24 -stimulated CREB phosphorylation, and steroidogenesis. Conclusions: Hypoglycosylated FSH21/18 has greater bioactivity than fully glycosylated hFSH24, suggesting that age-dependent decreases in hypoglycosylated hFSH contribute to reduced ovarian responsiveness. Hypoglycosylated FSH may be useful in follicle stimulation protocols for older patients using assisted reproduction technologies.

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U2 - 10.1210/jc.2015-1317

DO - 10.1210/jc.2015-1317

M3 - Article

VL - 100

SP - E852-E860

JO - Journal of Clinical Endocrinology and Metabolism

JF - Journal of Clinical Endocrinology and Metabolism

SN - 0021-972X

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