Heat shock protein 70 regulates platelet integrin activation, granule secretion and aggregation

Rachel A. Rigg, Laura D. Healy, Marie S. Nowak, Jérémy Mallet, Marisa L D Thierheimer, Jiaqing Pang, Owen McCarty, Joseph Aslan

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Molecular chaperones that support protein quality control, including heat shock protein 70 (Hsp70), participate in diverse aspects of cellular and physiological function. Recent studies have reported roles for specific chaperone activities in blood platelets in maintaining hemostasis; however, the functions of Hsp70 in platelet physiology remain uninvestigated. Here we characterize roles for Hsp70 activity in platelet activation and function. In vitro biochemical, microscopy, flow cytometry, and aggregometry assays of platelet function, as well as ex vivo analyses of platelet aggregate formation in whole blood under shear, were carried out under Hsp70-inhibited conditions. Inhibition of platelet Hsp70 blocked platelet aggregation and granule secretion in response to collagen-related peptide (CRP), which engages the immunoreceptor tyrosine-based activation motif-bearing collagen receptor glycoprotein VI (GPVI)-Fc receptor-γ chain complex. Hsp70 inhibition also reduced platelet integrin-αIIbβ3 activation downstream of GPVI, as Hsp70-inhibited platelets showed reduced PAC-1 and fibrinogen binding. Ex vivo, pharmacological inhibition of Hsp70 in human whole blood prevented the formation of platelet aggregates on collagen under shear. Biochemical studies supported a role for Hsp70 in maintaining the assembly of the linker for activation of T cells signalosome, which couples GPVI-initiated signaling to integrin activation, secretion, and platelet function. Together, our results suggest that Hsp70 regulates platelet activation and function by supporting linker for activation of T cells-associated signaling events downstream of platelet GPVI engagement, suggesting a role for Hsp70 in the intracellular organization of signaling systems that mediate platelet secretion, “inside-out” activation of platelet integrin- αIIbβ3, platelet- platelet aggregation, and, ultimately, hemostatic plug and thrombus formation.

Original languageEnglish (US)
Pages (from-to)C568-C575
JournalAmerican Journal of Physiology - Cell Physiology
Volume310
Issue number7
DOIs
StatePublished - Apr 1 2016

Fingerprint

HSP70 Heat-Shock Proteins
Platelet Activation
Integrins
Blood Platelets
Glycoproteins
Platelet Aggregation
Immunoreceptor Tyrosine-Based Activation Motif
Collagen Receptors
T-Lymphocytes
Molecular Chaperones
Fc Receptors
Hemostatics
Hemostasis
Quality Control
Fibrinogen
Microscopy
Flow Cytometry
Thrombosis
Collagen
Pharmacology

Keywords

  • Chaperones
  • Hemostasis
  • Integrin
  • Platelets
  • Thrombosis

ASJC Scopus subject areas

  • Physiology
  • Medicine(all)
  • Cell Biology

Cite this

Heat shock protein 70 regulates platelet integrin activation, granule secretion and aggregation. / Rigg, Rachel A.; Healy, Laura D.; Nowak, Marie S.; Mallet, Jérémy; Thierheimer, Marisa L D; Pang, Jiaqing; McCarty, Owen; Aslan, Joseph.

In: American Journal of Physiology - Cell Physiology, Vol. 310, No. 7, 01.04.2016, p. C568-C575.

Research output: Contribution to journalArticle

Rigg, Rachel A. ; Healy, Laura D. ; Nowak, Marie S. ; Mallet, Jérémy ; Thierheimer, Marisa L D ; Pang, Jiaqing ; McCarty, Owen ; Aslan, Joseph. / Heat shock protein 70 regulates platelet integrin activation, granule secretion and aggregation. In: American Journal of Physiology - Cell Physiology. 2016 ; Vol. 310, No. 7. pp. C568-C575.
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AU - Rigg, Rachel A.

AU - Healy, Laura D.

AU - Nowak, Marie S.

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AU - Thierheimer, Marisa L D

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AB - Molecular chaperones that support protein quality control, including heat shock protein 70 (Hsp70), participate in diverse aspects of cellular and physiological function. Recent studies have reported roles for specific chaperone activities in blood platelets in maintaining hemostasis; however, the functions of Hsp70 in platelet physiology remain uninvestigated. Here we characterize roles for Hsp70 activity in platelet activation and function. In vitro biochemical, microscopy, flow cytometry, and aggregometry assays of platelet function, as well as ex vivo analyses of platelet aggregate formation in whole blood under shear, were carried out under Hsp70-inhibited conditions. Inhibition of platelet Hsp70 blocked platelet aggregation and granule secretion in response to collagen-related peptide (CRP), which engages the immunoreceptor tyrosine-based activation motif-bearing collagen receptor glycoprotein VI (GPVI)-Fc receptor-γ chain complex. Hsp70 inhibition also reduced platelet integrin-αIIbβ3 activation downstream of GPVI, as Hsp70-inhibited platelets showed reduced PAC-1 and fibrinogen binding. Ex vivo, pharmacological inhibition of Hsp70 in human whole blood prevented the formation of platelet aggregates on collagen under shear. Biochemical studies supported a role for Hsp70 in maintaining the assembly of the linker for activation of T cells signalosome, which couples GPVI-initiated signaling to integrin activation, secretion, and platelet function. Together, our results suggest that Hsp70 regulates platelet activation and function by supporting linker for activation of T cells-associated signaling events downstream of platelet GPVI engagement, suggesting a role for Hsp70 in the intracellular organization of signaling systems that mediate platelet secretion, “inside-out” activation of platelet integrin- αIIbβ3, platelet- platelet aggregation, and, ultimately, hemostatic plug and thrombus formation.

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