Megakaryocyte Mechanosensing Toward Platelet Biogenesis

巨核细胞机械传感对血小板生物发生的影响

基本信息

批准号：
10473789
负责人：
KATYA RAVID
金额：
$ 41.25万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10473789
关键词：
Actins Address Adhesions Adult Affect Area Basement membrane Biogenesis Biology Blood Platelets Blood Vessels Bone Marrow Calcium Cations Cell physiology Cells Collagen Collagen Receptors Collagen Type IV Cytoskeleton Development Environment Extracellular Matrix Extracellular Matrix Proteins Family Fibronectin Receptors Fibronectins Generations Genetic Goals Greek Hemorrhage Hemostatic function Human Image In Vitro Integrins Intravenous Immunoglobulins Investigation Knock-out Knockout Mice Knowledge Life Marrow Measurement Mechanics Mechanoreceptors Mediating Megakaryocytes Mus Myelosuppression Neurons Outcome Pathology Pathway interactions Patients Pharmacology Phenotype Piezo 1 ion channel Piezo 2 ion channel Platelet Count measurement Platelet Transfusion Play Ploidies Production Property Proteins Receptor Activation Reporting Research Research Proposals Role Signal Transduction Testing Thrombocytopenia Thrombopoietin Thrombosis Transfusion Tubulin attenuation blood product crosslink experimental study high risk in vivo innovation insight leukemia mechanotransduction member mimetics mouse model novel preference pressure prevent receptor response side effect thrombocytosis thrombotic transcription factor translational potential

项目摘要

ABSTRACT Platelet counts are tightly regulated in order to prevent thrombotic or hemorrhagic complications associated with thrombocytosis or thrombocytopenia, respectively. While strides have been made in our understanding of proplatelet formation (PPF), there is still limited knowledge regarding the mechanisms through which the bone marrow (BM) extracellular matrix (ECM) regulates platelet production. Indeed, the BM includes a rich ECM with potential to generate mechanical constraints. Yet, the impact of BM mechanics on megakaryocyte (MK) properties and platelet formation has been understudied. Here, we propose an integrative approach to investigate emerging concepts related to the role of MK mechanobiological receptors in controlling MK adhesion to the ECM and platelet production. Our ultimate goal is to understand how specific MK mechanosensors sense the BM matrix to affect the cellular cytoskeleton, MK properties and, importantly, platelet level. Building upon our novel findings, Aim 1 explores the new paradigm and hypothesis that distinct MK cation channels preferentially respond to different BM matrix proteins and inversely impact the MK cytoskeleton and platelet levels. Experiments will focus on the Piezo family of cation channel mechanosensors, as compared to the Transient Receptor Potential cation channel subfamily V member 4 mechanosensor. In recent studies, we found these mechanosensors to have distinct preferences for different matrix proteins and opposing effects on PPF. Investigations will be carried out using pharmacological approaches as well as newly generated knockout mice at baseline and in response to challenges, such as myelosuppression or thrombocytopenia. Encouraged by preliminary studies using human primary MKs, continued studies will confirm murine findings. Aim 2 delineates mechanisms mediating novel connections between MK mechanosensors, integrin receptors activation, cytoskeletal changes, and MK mechano-sensitive transcription factors. This proposal is significant as there is need to identify new and alternative thrombopoietic pathways and agents that modulate platelet counts. In addition to conceptual innovation, at the technical level we will analyze new mouse models we developed with deletion of specific mechanosensors in MKs, and will apply state-of-the-art imaging and measurements under flow to follow cellular processes. Proposed studies are expected to yield new insights on the role of selective ECM sensing by MKs in controlling the MK cytoskeleton, adhesion and platelet production, with significant potential to impact our ability to modulate platelet levels.

抽象的严格控制血小板计数，以预防相关的血栓或出血并发症分别伴有血小板增多症或血小板减少症。虽然我们的理解已经取得了长足的进步前血小板形成（PPF），关于骨形成的机制仍然有限。骨髓 (BM) 细胞外基质 (ECM) 调节血小板的产生。事实上，BM 包含丰富的 ECM 产生机械约束的潜力。然而，BM力学对巨核细胞（MK）的影响性质和血小板形成已得到充分研究。在此，我们提出了一种综合方法研究与 MK 机械生物学受体在控制 MK 中的作用相关的新兴概念粘附 ECM 和血小板生成。我们的最终目标是了解具体的MK 机械传感器感知 BM 矩阵以影响细胞骨架、MK 特性，更重要的是，血小板水平。基于我们的新发现，目标 1 探索了新的范式和假设， MK 阳离子通道优先响应不同的 BM 基质蛋白并对 MK 产生反向影响细胞骨架和血小板水平。实验将集中在阳离子通道机械传感器的压电系列上，与瞬时受体电位阳离子通道亚家族 V 成员 4 机械传感器相比。在最近的研究，我们发现这些机械传感器对不同的基质蛋白有不同的偏好，对 PPF 的相反影响。将使用药理学方法以及新方法进行研究在基线和应对挑战（例如骨髓抑制或血小板减少症。受到使用人类初级 MK 的初步研究的鼓励，继续研究将确认小鼠的发现。目标 2 描绘了介导 MK 之间新连接的机制机械传感器、整合素受体激活、细胞骨架变化和 MK 机械敏感转录因素。该提案意义重大，因为需要确定新的和替代的血小板生成途径和调节血小板计数的药物。除了理念创新，在技术层面我们也会分析我们开发的删除 MK 中特定机械传感器的新小鼠模型，并将应用最先进的流动成像和测量，以跟踪细胞过程。拟议的研究是有望对 MK 选择性 ECM 传感在控制 MK 细胞骨架中的作用产生新的见解，粘附和血小板生成，具有显着影响我们调节血小板水平的能力的潜力。