The Biogenesis of Platelet-Derived Extracellular Vesicles and their Impact on Megakaryocyte Maturation

点击翻译按钮获取中文摘要

The Biogenesis of Platelet-Derived Extracellular Vesicles and their Impact on Megakaryocyte Maturation Abstract Thrombocytopenia is a major clinical problem encountered in multiple conditions, and severe thrombocytopenia (platelet counts <50 x 10^9/L) can lead to life threatening bleeding. Current treatment options have severe side effects, are in limited supply, involve blood products, and the platelet response typically takes up to 12 days. Therefore, there is an urgent need to identify new thrombopoietic agents that increase platelet counts for patients. In many inflammatory conditions platelet counts rise, resulting in thrombocytosis, but what initiates this platelet up-regulation is not well understood. Our lab uses inflammation as a model of exacerbated thrombopoiesis that results in differences in platelet quality and quantity in order to 1) gain a better understanding of the basic biology of megakaryocyte (MK) maturation their production of platelets, 2) identify thrombopoietin (TPO) independent pathways of MK maturation, and 3) determine ways to reduce platelet-related morbidity and mortality in inflammation. We have discovered a novel regulator of MK maturation during inflammation: platelet-derived extracellular vesicles (PEVs) in the bone marrow. Our preliminary data indicate that platelets package and shed MVs in an agonist-specific mechanism dependent on Rho GTPase signaling; the mechanism of Rho-mediated regulation of PEV formation and packaging will be explored in Aim 1. We also found that PEVs enter the bone marrow from the plasma, and bind to and are endocytosed by MKs both in vitro and in vivo. In Aim 2, we will examine how platelet-derived MVs interact with MKs. Specifically, we will determine the mechanisms by which they bind to and are internalized by MKs and how their cargo transferred. In inflammatory conditions such as SLE, ongoing platelet activation increases levels of circulating PEVs. These PEVs deliver disease-related changes from the plasma milieu directly to MKs in the bone marrow, reprogramming the MKs to make more pathogenic platelets. In Aim 3, we will identify the PEV factors that alter MK gene expression and platelet content in SLE. Successful completion of the proposed experiments will, for the first time, provide a detailed roadmap of how PEVs alter the hematopoietic environment in the setting of inflammatory disease. The insights gained may identify novel therapeutic targets that (i) alter PEV poduction independent of platelet activation (Aim 1), (ii) hijack the PEV/MK interaction to alter MK maturation (Aim 2), and (iii) inhibit pathologic MK reprogramming during SLE and other inflammatory diseases (Aim 3).