Uncovering the role of the Hsp40 family member cysteine string protein-α in mouse platelets

Abstract: Platelets modulate vascular microenvironments via the release of cargo molecules. Granule secretion is modulated by proteins called soluble N-ethylmaleimide sensitive factor attachment protein receptors (SNAREs). Secretion is complex and regulated by several protein-protein interactions; h...

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Main Authors: Alexis N. Smith, Hammodah R. Alfar, Smita Joshi, Daniëlle M. Coenen, Joshua Lykins, Isabelle C. Becker, Brittany E. Dong, Harry Chanzu, Chi Peng, Kanakanagavalli Shravani Prakhya, Linda Omali, Irina D. Pokrovskaya, Brian Storrie, Joseph Italiano, Sidney W. Whiteheart
Format: Article
Language:English
Published: Elsevier 2025-08-01
Series:Blood Advances
Online Access:http://www.sciencedirect.com/science/article/pii/S2473952925003179
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author Alexis N. Smith
Hammodah R. Alfar
Smita Joshi
Daniëlle M. Coenen
Joshua Lykins
Isabelle C. Becker
Brittany E. Dong
Harry Chanzu
Chi Peng
Kanakanagavalli Shravani Prakhya
Linda Omali
Irina D. Pokrovskaya
Brian Storrie
Joseph Italiano
Sidney W. Whiteheart
author_facet Alexis N. Smith
Hammodah R. Alfar
Smita Joshi
Daniëlle M. Coenen
Joshua Lykins
Isabelle C. Becker
Brittany E. Dong
Harry Chanzu
Chi Peng
Kanakanagavalli Shravani Prakhya
Linda Omali
Irina D. Pokrovskaya
Brian Storrie
Joseph Italiano
Sidney W. Whiteheart
author_sort Alexis N. Smith
collection DOAJ
description Abstract: Platelets modulate vascular microenvironments via the release of cargo molecules. Granule secretion is modulated by proteins called soluble N-ethylmaleimide sensitive factor attachment protein receptors (SNAREs). Secretion is complex and regulated by several protein-protein interactions; however, not all are characterized in platelets. We have identified cysteine string protein-α (CSPα; also known as, DNAJC5 or CLN4) as required for platelet secretion. CSPα is the only member from the CSP family present in platelets and has been proposed as a chaperone for the SNAP-23/25 t (Qb,c) SNAREs. To address CSPα’s role, we analyzed platelets from CSPα−/− mice. The loss of CSPα significantly affected dense- and α-granule release with minimal effects on lysosomal secretion. Consistent with the secretion defects, in vivo and ex vivo assays showed that loss of CSPα caused significant bleeding and attenuated thrombosis under flow. Interestingly, loss of CSPα caused a reduction in glycoprotein VI (GPVI) levels and reduced αIIbβ3 activation, especially in response to GPVI-specific agonists. Deletion of CSPα did not affect proteins in the platelet secretory machinery, for example, the SNAP-23/25 proteins. Subcellular fractionation studies showed that CSPα, which is reported to be acylated, was present on membranes but not in lipid rafts. Immunofluorescence studies showed CSPα colocalized with α and lysosomal granule markers. CSPα−/− mice had reduced red blood cell, leukocyte, and megakaryocyte numbers, suggesting effects on bone marrow progenitor cells. Simultaneously, we detected increased collagen I deposition, but no fibrosis in the marrow of CSPα−/− mice. These results identify CSPα as another element of the platelet secretory machinery that significantly contributes to thrombosis and hemostasis.
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spelling doaj-art-8a33304e7c3f4216b8ba10cab0761ac52025-08-20T04:00:55ZengElsevierBlood Advances2473-95292025-08-019164111412510.1182/bloodadvances.2024014870Uncovering the role of the Hsp40 family member cysteine string protein-α in mouse plateletsAlexis N. Smith0Hammodah R. Alfar1Smita Joshi2Daniëlle M. Coenen3Joshua Lykins4Isabelle C. Becker5Brittany E. Dong6Harry Chanzu7Chi Peng8Kanakanagavalli Shravani Prakhya9Linda Omali10Irina D. Pokrovskaya11Brian Storrie12Joseph Italiano13Sidney W. Whiteheart14Department of Molecular and Cellular Biochemistry, College of Medicine, University of Kentucky, Lexington, KYDepartment of Molecular and Cellular Biochemistry, College of Medicine, University of Kentucky, Lexington, KYDepartment of Molecular and Cellular Biochemistry, College of Medicine, University of Kentucky, Lexington, KYDepartment of Molecular and Cellular Biochemistry, College of Medicine, University of Kentucky, Lexington, KYDepartment of Molecular and Cellular Biochemistry, College of Medicine, University of Kentucky, Lexington, KYDepartment of Surgery, Boston Children’s Hospital, Boston, MADepartment of Molecular and Cellular Biochemistry, College of Medicine, University of Kentucky, Lexington, KYDepartment of Molecular and Cellular Biochemistry, College of Medicine, University of Kentucky, Lexington, KYDepartment of Molecular and Cellular Biochemistry, College of Medicine, University of Kentucky, Lexington, KYDepartment of Molecular and Cellular Biochemistry, College of Medicine, University of Kentucky, Lexington, KYDepartment of Molecular and Cellular Biochemistry, College of Medicine, University of Kentucky, Lexington, KYDepartment of Physiology and Biophysics, University of Arkansas for Medical Sciences, Little Rock, ARDepartment of Physiology and Biophysics, University of Arkansas for Medical Sciences, Little Rock, ARDepartment of Surgery, Boston Children’s Hospital, Boston, MADepartment of Molecular and Cellular Biochemistry, College of Medicine, University of Kentucky, Lexington, KY; Correspondence: Sidney W. Whiteheart, Department of Molecular and Cellular Biochemistry, College of Medicine, University of Kentucky, B361 BBSRB, 741 S Limestone, Lexington, KY 40536;Abstract: Platelets modulate vascular microenvironments via the release of cargo molecules. Granule secretion is modulated by proteins called soluble N-ethylmaleimide sensitive factor attachment protein receptors (SNAREs). Secretion is complex and regulated by several protein-protein interactions; however, not all are characterized in platelets. We have identified cysteine string protein-α (CSPα; also known as, DNAJC5 or CLN4) as required for platelet secretion. CSPα is the only member from the CSP family present in platelets and has been proposed as a chaperone for the SNAP-23/25 t (Qb,c) SNAREs. To address CSPα’s role, we analyzed platelets from CSPα−/− mice. The loss of CSPα significantly affected dense- and α-granule release with minimal effects on lysosomal secretion. Consistent with the secretion defects, in vivo and ex vivo assays showed that loss of CSPα caused significant bleeding and attenuated thrombosis under flow. Interestingly, loss of CSPα caused a reduction in glycoprotein VI (GPVI) levels and reduced αIIbβ3 activation, especially in response to GPVI-specific agonists. Deletion of CSPα did not affect proteins in the platelet secretory machinery, for example, the SNAP-23/25 proteins. Subcellular fractionation studies showed that CSPα, which is reported to be acylated, was present on membranes but not in lipid rafts. Immunofluorescence studies showed CSPα colocalized with α and lysosomal granule markers. CSPα−/− mice had reduced red blood cell, leukocyte, and megakaryocyte numbers, suggesting effects on bone marrow progenitor cells. Simultaneously, we detected increased collagen I deposition, but no fibrosis in the marrow of CSPα−/− mice. These results identify CSPα as another element of the platelet secretory machinery that significantly contributes to thrombosis and hemostasis.http://www.sciencedirect.com/science/article/pii/S2473952925003179
spellingShingle Alexis N. Smith
Hammodah R. Alfar
Smita Joshi
Daniëlle M. Coenen
Joshua Lykins
Isabelle C. Becker
Brittany E. Dong
Harry Chanzu
Chi Peng
Kanakanagavalli Shravani Prakhya
Linda Omali
Irina D. Pokrovskaya
Brian Storrie
Joseph Italiano
Sidney W. Whiteheart
Uncovering the role of the Hsp40 family member cysteine string protein-α in mouse platelets
Blood Advances
title Uncovering the role of the Hsp40 family member cysteine string protein-α in mouse platelets
title_full Uncovering the role of the Hsp40 family member cysteine string protein-α in mouse platelets
title_fullStr Uncovering the role of the Hsp40 family member cysteine string protein-α in mouse platelets
title_full_unstemmed Uncovering the role of the Hsp40 family member cysteine string protein-α in mouse platelets
title_short Uncovering the role of the Hsp40 family member cysteine string protein-α in mouse platelets
title_sort uncovering the role of the hsp40 family member cysteine string protein α in mouse platelets
url http://www.sciencedirect.com/science/article/pii/S2473952925003179
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