Platelet granule formation and function in health and disease

血小板颗粒的形成及其在健康和疾病中的功能

• 批准号: 9257459
• 负责人: Michael S Marks
• 金额: $ 47.56万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-07 至 2020-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9257459
• 关键词: Adhesions; Affect; Alpha Granule; Biogenesis; Blood Coagulation Disorders; Blood Platelet Disorders; Blood Platelets; Blood Vessels; Bone Marrow; Calcium; Cell membrane; Cells; Chimeric Proteins; Coagulation Process; Complex; Cytoplasmic Granules; Data; Defect; Diagnostic; Disease; Disease susceptibility; Early Endosome; Endosomes; Epitopes; Fetal Liver; Genes; Health; Hemorrhage; Hemorrhagic Disorders; Hemostatic function; Hereditary Disease; Hermanski-Pudlak Syndrome; Human; Impairment; In Situ; Inflammation; Integral Membrane Protein; Label; Lysosomes; Megakaryocytes; Melanosomes; Membrane; Membrane Proteins; Modeling; Molecular; Morphology; Mus; Oculocutaneous Albinism; Organelles; Pathology; Pathway interactions; Patients; Pharmacology; Phenotype; Pigments; Process; Proteins; Quinacrine; Role; Secondary to; Serotonin; Sorting - Cell Movement; Source; Structure; Testing; Therapeutic; Therapeutic Intervention; Thrombus; Tissues; Transport Process; Variant; Vesicle; Vesicle Transport Pathway; Wound Healing; angiogenesis; base; cell type; cohort; functional disability; improved; in vivo; induced pluripotent stem cell; insight; melanocyte; mouse model; protein complex; public health relevance; reconstitution; small molecule; tool

DESCRIPTION (provided by applicant): Platelets fulfill essential functions in hemostasis, inflammation, angiogenesis, and other processes. Many of these functions require regulated secretion from three types of storage compartments - � granules, dense granules, and lysosomes. Many hemorrhagic and thrombotic disorders are caused by dysregulation of granule formation and/or content release. In particular, bleeding diathesis in Hermansky-Pudlak syndrome (HPS) results from an absence of detectable platelet dense granules and consequent defects in activation-dependent release of calcium and ADP, which normally promote thrombus formation. Despite the importance of platelet granules, the mechanisms underlying their formation and membrane dynamics within megakaryocytes (MKs) and/or their derived platelets are largely uncharacterized, and the pathways regulated by the genes that are defective in the nine characterized HPS variants are not known. In other cell types, HPS-associated genes regulate vesicular transport processes required to deliver resident integral membrane proteins from early endosomes to lysosome-related organelles - cell type-specific intracellular storage compartments like � and dense granules. We and our collaborators have recently identified two integral membrane proteins - SLC35D3 and VMAT2 - as candidate dense granule-specific cargoes that can serve as markers to study dense granule biogenesis. Surprisingly, these two cargoes localize to early endosomes in MKs from both wild-type and HPS model mice. Nevertheless, SLC35D3 is destabilized in platelets from the same HPS models. Based on these and other data, we hypothesize that dense granule integral membrane proteins are delivered from early endosomes to immature dense granules at a late stage of differentiation of platelets from MKs. We further hypothesize that dense granule membranes are not static but are constantly remodeled in active platelets, perhaps providing an avenue for delivery of pharmacological agents to platelet dense granules. Finally, preliminary data suggest that activation-induced content release is impaired not only from dense granules but also from � granules and lysosomes in HPS model platelets, both in vivo and ex vivo. Since content release requires fusion of the granule membrane with the plasma membrane, we hypothesize that this secretory defect in HPS models reflects impaired cargo delivery of the fusion machinery to � granules and lysosomes. We will test these hypotheses in the following Specific Aims: 1. To assess whether HPS-associated proteins regulate the dynamic localization of candidate dense granule membrane proteins in platelets. 2. To assess whether HPS impacts maturation of dense granules by fusion of distinct precursor organelles. 3. To test whether and how HPS subtypes impact �-granule and lysosome secretion from platelets.

描述（由申请人提供）：血小板在止血、炎症、血管生成和其他过程中发挥重要作用。许多这些功能需要三种类型的储存室——颗粒、致密颗粒和溶酶体的调节分泌。许多出血性和血栓性疾病是由颗粒形成和/或内容物释放的失调引起的。特别是，Hermansky-Pudlak综合征（HPS）的出血特征是由于缺乏可检测到的血小板致密颗粒，以及由此导致的钙和ADP的激活依赖性释放缺陷，而钙和ADP通常会促进血栓的形成。尽管血小板颗粒很重要，但它们在巨核细胞（mk）和/或其衍生的血小板内形成和膜动力学的机制在很大程度上是未知的，并且在九种特征的HPS变异中由缺陷基因调节的途径尚不清楚。在其他类型的细胞中，hps相关基因调节将常驻整体膜蛋白从早期内体运送到溶酶体相关细胞器（细胞类型特异性细胞内存储室，如致密颗粒）所需的囊泡运输过程。我们和我们的合作者最近确定了两个完整的膜蛋白- SLC35D3和VMAT2 -作为候选的致密颗粒特异性货物，可以作为研究致密颗粒生物发生的标记。令人惊讶的是，这两种货物都定位于野生型和HPS模型小鼠mk的早期核内体。然而，SLC35D3在相同HPS模型的血小板中是不稳定的。基于这些和其他数据，我们假设在血小板从mk分化的后期，致密颗粒整体膜蛋白从早期内体传递到未成熟的致密颗粒。我们进一步假设致密颗粒膜不是静态的，而是在活性血小板中不断重塑的，这可能为药物输送到血小板致密颗粒提供了途径。最后，初步数据表明，在体内和离体实验中，HPS模型血小板中活化诱导的内容物释放不仅受到致密颗粒的损害，也受到颗粒和溶酶体的损害。由于内容物释放需要颗粒膜与质膜融合，我们假设HPS模型的这种分泌缺陷反映了融合机制对颗粒和溶酶体的货物输送受损。我们将在以下具体目标中检验这些假设：评估hps相关蛋白是否调节血小板中候选致密颗粒膜蛋白的动态定位。2. 评估HPS是否通过不同前体细胞的融合影响致密颗粒的成熟。3. 测试HPS亚型是否以及如何影响血小板的颗粒和溶酶体分泌。

项目成果

Tyrosine 870 of TLR9 is critical for receptor maturation rather than phosphorylation-dependent ligand-induced signaling.

• DOI: 10.1371/journal.pone.0200913
• 发表时间: 2018
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Biswas C;Rao S;Slade K;Hyman D;Dersh D;Mantegazza AR;Zoltick PW;Marks MS;Argon Y;Behrens EM
• 通讯作者: Behrens EM

Pink Light on Mitochondria in Autoimmunity and Parkinson Disease.

自身免疫和帕金森病线粒体上的粉红光。

• DOI: 10.1016/j.cmet.2016.06.022
• 发表时间: 2016
• 期刊: Cell metabolism
• 影响因子: 29
• 作者: Mantegazza,AdrianaR;Marks,MichaelS
• 通讯作者: Marks,MichaelS