Targeting cargo to an unconventional secretory system within megakaryocytes allows the release of transgenic proteins from platelets.

Megakaryocyte bone marrow dense tubular system granules hemostasis platelet proplatelet protein disulfide isomerase thrombosis

Journal

Journal of thrombosis and haemostasis : JTH
ISSN: 1538-7836
Titre abrégé: J Thromb Haemost
Pays: England
ID NLM: 101170508

Informations de publication

Date de publication:
07 Aug 2024
Historique:
received: 16 01 2024
revised: 21 07 2024
accepted: 23 07 2024
medline: 10 8 2024
pubmed: 10 8 2024
entrez: 9 8 2024
Statut: aheadofprint

Résumé

Platelets are essential for hemostasis, and thrombosis, and play vital roles during metastatic cancer progression and infection. Hallmarks of platelet function are activation, cytoskeletal rearrangements, and the degranulation of their cellular contents upon stimulation. While alpha and dense granules are the most studied platelet secretory granules, the dense tubular system (DTS) also functions as a secretory system for vascular thiol isomerases. However, how DTS cargo is packaged and transported from megakaryocytes (MKs) to platelets is poorly understood. To underpin the mechanisms responsible for DTS cargo transport and leverage those for therapeutic protein packaging into platelets. A retroviral expression system combined with immunofluorescence confocal microscopy was employed to track protein DTS cargo protein disulfide isomerase (PDI) fused to eGFP (eGFP-PDI) during platelet production. Murine bone marrow transplantation models were used to determine the release of therapeutic proteins from platelets. We demonstrated that the endoplasmic reticulum retrieval motif Lys-Asp-Glu-Leu (KDEL) located at the C-terminus of PDI was essential for the regular transport of eGFP-PDI-containing granules. eGFP-PDI

Sections du résumé

BACKGROUND BACKGROUND
Platelets are essential for hemostasis, and thrombosis, and play vital roles during metastatic cancer progression and infection. Hallmarks of platelet function are activation, cytoskeletal rearrangements, and the degranulation of their cellular contents upon stimulation. While alpha and dense granules are the most studied platelet secretory granules, the dense tubular system (DTS) also functions as a secretory system for vascular thiol isomerases. However, how DTS cargo is packaged and transported from megakaryocytes (MKs) to platelets is poorly understood.
OBJECTIVES OBJECTIVE
To underpin the mechanisms responsible for DTS cargo transport and leverage those for therapeutic protein packaging into platelets.
METHODS METHODS
A retroviral expression system combined with immunofluorescence confocal microscopy was employed to track protein DTS cargo protein disulfide isomerase (PDI) fused to eGFP (eGFP-PDI) during platelet production. Murine bone marrow transplantation models were used to determine the release of therapeutic proteins from platelets.
RESULTS AND CONCLUSIONS CONCLUSIONS
We demonstrated that the endoplasmic reticulum retrieval motif Lys-Asp-Glu-Leu (KDEL) located at the C-terminus of PDI was essential for the regular transport of eGFP-PDI-containing granules. eGFP-PDI

Identifiants

DOI: 10.1016/j.jtha.2024.07.021 PMID: 39122192
pubmed: 39122192
pii: S1538-7836(24)00440-9
doi: 10.1016/j.jtha.2024.07.021
pii:
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Informations de copyright

Copyright © 2024 International Society on Thrombosis and Haemostasis. Published by Elsevier Inc. All rights reserved.

Auteurs

Nathan L Asquith (NL)

Vascular Biology Program, Boston Children's Hospital, 1 Blackfan Circle, Karp Research Building, Boston, MA, USA; Harvard Medical School, Boston, MA, USA.

Isabelle C Becker (IC)

Vascular Biology Program, Boston Children's Hospital, 1 Blackfan Circle, Karp Research Building, Boston, MA, USA; Harvard Medical School, Boston, MA, USA.

Mark T Scimone (MT)

Cellular Imaging Core, Neurobiology, Boston Children's Hospital 3 Blackfan Circle, Boston, MA, USA; Life Sciences, Biotechnology, University of New Hampshire, Manchester, NH, USA.

Thais Boccia Da Costa (TB)

Department of Surgery, Boston Children's Hospital, 1 Blackfan Circle, Karp Research Building, Boston, MA, USA; Harvard Medical School, Boston, MA, USA.

Virginia Camacho (V)

Vascular Biology Program, Boston Children's Hospital, 1 Blackfan Circle, Karp Research Building, Boston, MA, USA; Harvard Medical School, Boston, MA, USA.

María N Barrachina (MN)

Vascular Biology Program, Boston Children's Hospital, 1 Blackfan Circle, Karp Research Building, Boston, MA, USA; Harvard Medical School, Boston, MA, USA.

Shihui Guo (S)

Harvard Medical School, Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center, 3 Blackfan Circle, Boston, MA, USA.

Daniela Freire (D)

Vascular Biology Program, Boston Children's Hospital, 1 Blackfan Circle, Karp Research Building, Boston, MA, USA.

Kellie Machlus (K)

Vascular Biology Program, Boston Children's Hospital, 1 Blackfan Circle, Karp Research Building, Boston, MA, USA; Harvard Medical School, Boston, MA, USA.

Sol Schulman (S)

Harvard Medical School, Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center, 3 Blackfan Circle, Boston, MA, USA.

Robert Flaumenhaft (R)

Harvard Medical School, Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center, 3 Blackfan Circle, Boston, MA, USA.

Joseph E Italiano (JE)

Vascular Biology Program, Boston Children's Hospital, 1 Blackfan Circle, Karp Research Building, Boston, MA, USA; Harvard Medical School, Boston, MA, USA. Electronic address: Joseph.Italiano@childrens.harvard.edu.

Classifications MeSH