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

基本信息

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
项目类别：
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

项目摘要

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功能，其中许多功能需要三种类型的储存室的调节分泌——颗粒、致密颗粒和溶酶体。是由颗粒形成和/或内容物释放失调引起的，特别是赫曼斯基-普德拉克综合征（HPS）的出血素质。由于缺乏可检测到的血小板致密颗粒以及随之而来的钙和 ADP 的激活依赖性释放缺陷，这些缺陷通常会促进血栓形成，尽管血小板颗粒很重要，但巨核细胞 (MK) 和/或内其形成和膜动力学的机制却很重要。它们衍生的血小板在很大程度上是未知的，并且九种特征性 HPS 变体中缺陷基因所调节的途径尚不清楚。在其他细胞类型中，HPS 相关基因调节囊泡运输过程。将驻留的完整膜蛋白从早期内体传递到溶酶体相关的细胞器（细胞类型特异性的细胞内储存室，例如和致密颗粒）我们和我们的合作者最近确定了两种完整膜蛋白 - SLC35D3 和 VMAT2 - 作为候选致密颗粒。令人惊讶的是，这两种货物定位于野生型和 HPS 模型中 MK 的早期内体。然而，SLC35D3 在来自相同 HPS 模型的血小板中不稳定。基于这些和其他数据，我们发现致密颗粒整合膜蛋白在血小板从 MK 分化的后期从早期内体传递到未成熟的致密颗粒。我们进一步发现，致密颗粒膜不是静态的，而是在活性血小板中不断重塑，这可能为向血小板致密颗粒输送药物提供了途径。最后，初步数据。表明激活诱导的内容物释放不仅受到致密颗粒的损害，而且受到 HPS 模型血小板中颗粒和溶酶体的损害，无论是在体内还是离体，由于内容物释放需要颗粒膜与质膜的融合，因此我们很高兴这一点。 HPS 模型中的这种分泌缺陷反映了融合机制向颗粒和溶酶体的货物递送受损。我们将在以下具体目标中测试这些假设： 1. 评估 HPS 相关蛋白是否存在。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器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 β 颗粒和溶酶体分泌。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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。