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Nascent preplatelets and B4GALT1 glycosylation contribute to 5-FU-induced bone marrow recovery
Natalia Weich1, Leonardo Rivadeneyra1, Alejandro Roisman1
1Translational Glycomics Center, Versiti Blood Research Institute, Milwaukee, WI.
Abstract:
Megakaryocytes (MKs) are essential for hemostasis, vascular integrity, and hematopoietic stem and progenitor cell (HSPC) support, but their role in bone marrow (BM) recovery has remained underexplored. Here, using a 5-fluorouracil (5-FU) injury model, we identify a transient intramedullary space enriched in extracellular matrix (ECM) proteins such as perlecan, von Willebrand factor, and heparanase, as well as functionally responsive GPIbα+ platelet particles (termed preplatelets). During 5-FU-induced injury, this compartment undergoes dynamic changes, and its resolution depends on the presence of functional preplatelets and ECM components. Consistent with this, platelet depletion further delays restoration of the intramedullary space despite preserved MK numbers, supporting a critical local role for nascent platelets during 5-FU-induced injury. Furthermore, mice lacking β-1,4-galactosyltransferase 1 (B4GALT1) develop persistent thrombocytopenia, exhibit mislocalized and morphologically abnormal MKs, and display expansion of MK-biased HSPCs after 5-FU injury, collectively leading to delayed hematopoietic recovery and expansion of the BM intramedullary space. Single-cell transcriptomic analysis of B4GALT1-/- MK-biased HSPCs at steady state further revealed disruption of adhesion, cytoskeletal, and Notch1-associated programs required for proplatelet formation and MK interactions with ECM components. Our findings reveal a previously unrecognized role for locally retained platelet intermediates and identify B4GALT1-dependent glycosylation as a key regulator of megakaryocyte integrity, platelet production, and hematopoietic recovery after 5-FU-induced myeloablation.
Insights
Newly discovered preplatelets and extracellular matrix proteins aid bone marrow recovery after injury. The enzyme B4GALT1 is crucial for megakaryocyte function and platelet production, impacting hematopoietic recovery.
Area of Science:
- Hematology
- Cell Biology
- Biochemistry
Background:
- Megakaryocytes (MKs) are vital for hemostasis, vascular integrity, and supporting hematopoietic stem and progenitor cells (HSPCs).
- The role of MKs and their products in bone marrow (BM) recovery following injury is not well understood.
- A 5-fluorouracil (5-FU) model is used to investigate BM recovery mechanisms.
Purpose of the Study:
- To identify the role of megakaryocytes and platelet intermediates in bone marrow recovery after chemotherapy-induced injury.
- To investigate the function of B4GALT1 in megakaryocyte integrity and platelet production.
- To understand the molecular mechanisms underlying hematopoietic recovery.
Main Methods:
- Utilized a 5-fluorouracil (5-FU) induced bone marrow injury model in mice.
- Analyzed the intramedullary space composition and dynamics during injury and recovery.
- Employed platelet depletion and genetic knockout (B4GALT1⁻/⁻) strategies.
- Performed single-cell transcriptomic analysis on MK-biased HSPCs.
Main Results:
- Identified a transient intramedullary space containing extracellular matrix (ECM) proteins and preplatelets, crucial for BM recovery.
- Platelet depletion delayed intramedullary space restoration, highlighting the role of nascent platelets.
- Mice lacking B4GALT1 showed persistent thrombocytopenia, abnormal MKs, and delayed hematopoietic recovery.
- B4GALT1 deficiency disrupted MK-biased HSPC programs related to adhesion, cytoskeleton, and Notch1 signaling.
Conclusions:
- Locally retained platelet intermediates and ECM components play a critical, previously unrecognized role in bone marrow recovery.
- B4GALT1-dependent glycosylation is essential for maintaining megakaryocyte integrity and platelet production.
- Proper MK function and platelet generation are key determinants of hematopoietic recovery after myeloablation.
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