VAMP8/endobrevin is overexpressed in hyperreactive human platelets

Suggested role for platelet microRNA

A. A. Kondkar, M. S. Bray, S. M. Leal, S. Nagalla, Dajiang Liu, Y. Jin, J. F. Dong, Q. Ren, S. W. Whiteheart, C. Shaw, P. F. Bray

Research output: Contribution to journalArticle

124 Citations (Scopus)

Abstract

Background: Variation in platelet reactivity contributes to disorders of hemostasis and thrombosis, but the molecular mechanisms are not well understood. Objectives: To discover associations between interindividual platelet variability and the responsible platelet genes, and to begin to define the molecular mechanisms altering platelet gene expression.Subjects/methods: Two hundred and eighty-eight healthy subjects were phenotyped for platelet responsiveness. Platelet RNA from subjects demonstrating hyperreactivity (n = 18) and hyporeactivity (n = 11) was used to screen the human transcriptome. Results: Distinctly different mRNA profiles were observed between subjects with differing platelet reactivity. Increased levels of mRNA for VAMP8/endobrevin, a critical v-SNARE involved in platelet granule secretion, were associated with platelet hyperreactivity (Q = 0.0275). Validation studies of microarray results showed 4.8-fold higher mean VAMP8 mRNA levels in hyperreactive than hyporeactive platelets (P = 0.0023). VAMP8 protein levels varied 13-fold among platelets from these normal subjects, and were 2.5-fold higher in hyperreactive platelets (P = 0.05). Among our cohort of 288 subjects, a VAMP8 single-nucleotide polymorphism (rs1010) was associated with platelet reactivity in an age-dependent manner (P < 0.003). MicroRNA-96 was predicted to bind to the 3′-untranslated regionof VAMP8 mRNA and was detected in platelets. Overexpression of microRNA-96 in VAMP8-expressing cell lines caused a dose-dependent decrease in VAMP8 protein and mRNA, suggesting a role in VAMP8 mRNA degradation. Conclusions:. These findings support a role for VAMP8/endobrevin in the heterogeneity of platelet reactivity, and suggest a role for microRNA-96 in the regulation of VAMP8 expression.

Original languageEnglish (US)
Pages (from-to)369-378
Number of pages10
JournalJournal of Thrombosis and Haemostasis
Volume8
Issue number2
DOIs
StatePublished - Feb 1 2010

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MicroRNAs
Blood Platelets
Messenger RNA
SNARE Proteins
Validation Studies
RNA Stability
Hemostasis
Transcriptome
Single Nucleotide Polymorphism
Healthy Volunteers
Proteins
Thrombosis

All Science Journal Classification (ASJC) codes

  • Hematology

Cite this

Kondkar, A. A. ; Bray, M. S. ; Leal, S. M. ; Nagalla, S. ; Liu, Dajiang ; Jin, Y. ; Dong, J. F. ; Ren, Q. ; Whiteheart, S. W. ; Shaw, C. ; Bray, P. F. / VAMP8/endobrevin is overexpressed in hyperreactive human platelets : Suggested role for platelet microRNA. In: Journal of Thrombosis and Haemostasis. 2010 ; Vol. 8, No. 2. pp. 369-378.
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abstract = "Background: Variation in platelet reactivity contributes to disorders of hemostasis and thrombosis, but the molecular mechanisms are not well understood. Objectives: To discover associations between interindividual platelet variability and the responsible platelet genes, and to begin to define the molecular mechanisms altering platelet gene expression.Subjects/methods: Two hundred and eighty-eight healthy subjects were phenotyped for platelet responsiveness. Platelet RNA from subjects demonstrating hyperreactivity (n = 18) and hyporeactivity (n = 11) was used to screen the human transcriptome. Results: Distinctly different mRNA profiles were observed between subjects with differing platelet reactivity. Increased levels of mRNA for VAMP8/endobrevin, a critical v-SNARE involved in platelet granule secretion, were associated with platelet hyperreactivity (Q = 0.0275). Validation studies of microarray results showed 4.8-fold higher mean VAMP8 mRNA levels in hyperreactive than hyporeactive platelets (P = 0.0023). VAMP8 protein levels varied 13-fold among platelets from these normal subjects, and were 2.5-fold higher in hyperreactive platelets (P = 0.05). Among our cohort of 288 subjects, a VAMP8 single-nucleotide polymorphism (rs1010) was associated with platelet reactivity in an age-dependent manner (P < 0.003). MicroRNA-96 was predicted to bind to the 3′-untranslated regionof VAMP8 mRNA and was detected in platelets. Overexpression of microRNA-96 in VAMP8-expressing cell lines caused a dose-dependent decrease in VAMP8 protein and mRNA, suggesting a role in VAMP8 mRNA degradation. Conclusions:. These findings support a role for VAMP8/endobrevin in the heterogeneity of platelet reactivity, and suggest a role for microRNA-96 in the regulation of VAMP8 expression.",
author = "Kondkar, {A. A.} and Bray, {M. S.} and Leal, {S. M.} and S. Nagalla and Dajiang Liu and Y. Jin and Dong, {J. F.} and Q. Ren and Whiteheart, {S. W.} and C. Shaw and Bray, {P. F.}",
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Kondkar, AA, Bray, MS, Leal, SM, Nagalla, S, Liu, D, Jin, Y, Dong, JF, Ren, Q, Whiteheart, SW, Shaw, C & Bray, PF 2010, 'VAMP8/endobrevin is overexpressed in hyperreactive human platelets: Suggested role for platelet microRNA', Journal of Thrombosis and Haemostasis, vol. 8, no. 2, pp. 369-378. https://doi.org/10.1111/j.1538-7836.2009.03700.x

VAMP8/endobrevin is overexpressed in hyperreactive human platelets : Suggested role for platelet microRNA. / Kondkar, A. A.; Bray, M. S.; Leal, S. M.; Nagalla, S.; Liu, Dajiang; Jin, Y.; Dong, J. F.; Ren, Q.; Whiteheart, S. W.; Shaw, C.; Bray, P. F.

In: Journal of Thrombosis and Haemostasis, Vol. 8, No. 2, 01.02.2010, p. 369-378.

Research output: Contribution to journalArticle

TY - JOUR

T1 - VAMP8/endobrevin is overexpressed in hyperreactive human platelets

T2 - Suggested role for platelet microRNA

AU - Kondkar, A. A.

AU - Bray, M. S.

AU - Leal, S. M.

AU - Nagalla, S.

AU - Liu, Dajiang

AU - Jin, Y.

AU - Dong, J. F.

AU - Ren, Q.

AU - Whiteheart, S. W.

AU - Shaw, C.

AU - Bray, P. F.

PY - 2010/2/1

Y1 - 2010/2/1

N2 - Background: Variation in platelet reactivity contributes to disorders of hemostasis and thrombosis, but the molecular mechanisms are not well understood. Objectives: To discover associations between interindividual platelet variability and the responsible platelet genes, and to begin to define the molecular mechanisms altering platelet gene expression.Subjects/methods: Two hundred and eighty-eight healthy subjects were phenotyped for platelet responsiveness. Platelet RNA from subjects demonstrating hyperreactivity (n = 18) and hyporeactivity (n = 11) was used to screen the human transcriptome. Results: Distinctly different mRNA profiles were observed between subjects with differing platelet reactivity. Increased levels of mRNA for VAMP8/endobrevin, a critical v-SNARE involved in platelet granule secretion, were associated with platelet hyperreactivity (Q = 0.0275). Validation studies of microarray results showed 4.8-fold higher mean VAMP8 mRNA levels in hyperreactive than hyporeactive platelets (P = 0.0023). VAMP8 protein levels varied 13-fold among platelets from these normal subjects, and were 2.5-fold higher in hyperreactive platelets (P = 0.05). Among our cohort of 288 subjects, a VAMP8 single-nucleotide polymorphism (rs1010) was associated with platelet reactivity in an age-dependent manner (P < 0.003). MicroRNA-96 was predicted to bind to the 3′-untranslated regionof VAMP8 mRNA and was detected in platelets. Overexpression of microRNA-96 in VAMP8-expressing cell lines caused a dose-dependent decrease in VAMP8 protein and mRNA, suggesting a role in VAMP8 mRNA degradation. Conclusions:. These findings support a role for VAMP8/endobrevin in the heterogeneity of platelet reactivity, and suggest a role for microRNA-96 in the regulation of VAMP8 expression.

AB - Background: Variation in platelet reactivity contributes to disorders of hemostasis and thrombosis, but the molecular mechanisms are not well understood. Objectives: To discover associations between interindividual platelet variability and the responsible platelet genes, and to begin to define the molecular mechanisms altering platelet gene expression.Subjects/methods: Two hundred and eighty-eight healthy subjects were phenotyped for platelet responsiveness. Platelet RNA from subjects demonstrating hyperreactivity (n = 18) and hyporeactivity (n = 11) was used to screen the human transcriptome. Results: Distinctly different mRNA profiles were observed between subjects with differing platelet reactivity. Increased levels of mRNA for VAMP8/endobrevin, a critical v-SNARE involved in platelet granule secretion, were associated with platelet hyperreactivity (Q = 0.0275). Validation studies of microarray results showed 4.8-fold higher mean VAMP8 mRNA levels in hyperreactive than hyporeactive platelets (P = 0.0023). VAMP8 protein levels varied 13-fold among platelets from these normal subjects, and were 2.5-fold higher in hyperreactive platelets (P = 0.05). Among our cohort of 288 subjects, a VAMP8 single-nucleotide polymorphism (rs1010) was associated with platelet reactivity in an age-dependent manner (P < 0.003). MicroRNA-96 was predicted to bind to the 3′-untranslated regionof VAMP8 mRNA and was detected in platelets. Overexpression of microRNA-96 in VAMP8-expressing cell lines caused a dose-dependent decrease in VAMP8 protein and mRNA, suggesting a role in VAMP8 mRNA degradation. Conclusions:. These findings support a role for VAMP8/endobrevin in the heterogeneity of platelet reactivity, and suggest a role for microRNA-96 in the regulation of VAMP8 expression.

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U2 - 10.1111/j.1538-7836.2009.03700.x

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