The Role of Striated Muscle Preferentially Expressed Protein Kinase in the Regulation of Ryanodine Receptor Type-2.

横纹肌优先表达的蛋白激酶在 2 型 Ryanodine 受体调节中的作用。

基本信息

  • 批准号:
    10006585
  • 负责人:
  • 金额:
    $ 4.21万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2018
  • 资助国家:
    美国
  • 起止时间:
    2018-09-01 至 2021-08-31
  • 项目状态:
    已结题

项目摘要

There is a fundamental gap in understanding of the mechanisms that regulate cardiac sarcoplasmic reticulum (SR) calcium release through Ryanodine Receptor Type-2 (RyR2). A fuller understanding of the regulation of RyR2 is needed to better comprehend mechanisms of calcium dysregulation in cardiovascular pathology. Striated Muscle Preferentially Expressed Protein Kinase (SPEG), a binding partner of RyR2, is a protein with dual kinase domains that is downregulated in heart failure. Cardiac loss of SPEG results in decreased SR stores through increased diastolic calcium leak from RyR2. Although its kinase activity has not been extensively studied, there is evidence that the internal kinase domain, SPEG1, is catalytically active. As previous phosphorylation events have been shown to modify RyR2 activity, it is possible that SPEG acts as a regulator of RyR2 through its kinase activity. The overall objective of this application is to determine what role SPEG’s kinase activity has on SR calcium handling and more specifically whether SPEG’s kinase activity is important in the regulation of RyR2. Preliminary data generated in the applicant’s laboratory revealed a significant decrease in phosphorylation at the RyR2-S2367 site in a cardiac specific SPEG knock-out mouse. Phosphorylation of this site, located in the critical clamp region of RyR2, has not been reported in the literature. The central hypothesis is that phosphorylation of the RyR2-S2367 site results in decreased diastolic calcium leak through stabilization of RyR2. The rationale for this research is that an understanding of the effects of this novel phosphorylation site and the mechanism of this phosphorylation event has the potential to lead to novel approaches to modulate RyR2 activity. The hypothesis will be tested with the following specific aims 1) Determine whether SPEG primarily phosphorylates RyR2 at the S2367 site, 2) Test the hypothesis that phosphorylation at the S2367 site inhibits diastolic RyR2 opening, and 3) Determine whether SPEG1 kinase activity is therapeutic in heart failure. For the first aim, we will determine whether RyR2-S2367 is phosphorylated by SPEG using both an in vitro kinase assay and overexpression of the SPEG1 kinase domain with adeno-associated virus. Under aim 2, we will utilize recombinant RyR2 and knock-in mouse models with RyR2-S2367 phospho-mimetic and phospho-resistant mutations to study the effects of this phosphorylation event on calcium release using both single channel RyR2 studies and calcium imaging with isolated cardiomyocytes. For aim 3, we will determine whether overexpression of the SPEG1 kinase domain with adeno-associated virus improves ejection fraction and SR calcium handling in a non-ischemic mouse model of heart failure. This research is significant in that it will advance understanding of a novel RyR2 phosphorylation site and the physiological importance of SPEG’s kinase activity. This knowledge has the potential to aid in the identification of novel therapeutic targets for the treatment of cardiovascular diseases such as heart failure.
对调节心脏肌浆的机制的理解存在根本差距 通过 Ryanodine 受体 2 型 (RyR2) 释放网状 (SR) 钙。更全面地了解 为了更好地理解心血管钙失调的机制,需要调节 RyR2 病理。横纹肌优先表达蛋白激酶 (SPEG) 是 RyR2 的结合伴侣,是一种 具有双激酶结构域的蛋白质,在心力衰竭中下调。 SPEG 心脏丧失导致 通过增加 RyR2 的舒张期钙泄漏来减少 SR 储存。尽管其激酶活性尚未 经过广泛研究,有证据表明内部激酶结构域 SPEG1 具有催化活性。作为 先前的磷酸化事件已被证明可以改变 RyR2 活性,SPEG 可能充当 通过其激酶活性调节 RyR2。此应用程序的总体目标是确定角色 SPEG 的激酶活性对 SR 钙处理有影响,更具体地说,SPEG 的激酶活性是否影响 对 RyR2 的调节很重要。申请人实验室生成的初步数据显示 心脏特异性 SPEG 敲除小鼠中 RyR2-S2367 位点的磷酸化显着降低。 该位点位于 RyR2 的关键钳区域,尚未见文献报道。 中心假设是 RyR2-S2367 位点磷酸化导致舒张期钙减少 通过稳定 RyR2 进行泄漏。这项研究的基本原理是了解这种影响 新的磷酸化位点和该磷酸化事件的机制有可能导致新的磷酸化位点 调节 RyR2 活性的方法。该假设将通过以下具体目标进行检验 1) 确定 SPEG 是否主要在 S2367 位点磷酸化 RyR2,2) 检验以下假设: S2367 位点抑制舒张 RyR2 打开,并且 3) 确定 SPEG1 激酶活性是否具有治疗作用 在心力衰竭中。对于第一个目标,我们将使用以下两种方法确定 RyR2-S2367 是否被 SPEG 磷酸化: 体外激酶测定以及腺相关病毒 SPEG1 激酶结构域的过表达。在下面 目标 2,我们将利用重组 RyR2 和带有 RyR2-S2367 磷酸化模拟物的敲入小鼠模型 磷酸化抗性突变,以研究这种磷酸化事件对钙释放的影响 单通道 RyR2 研究和离体心肌细胞的钙成像。对于目标 3,我们将确定 腺相关病毒过度表达 SPEG1 激酶结构域是否可以提高射血分数 SR 钙处理非缺血性心力衰竭小鼠模型。这项研究的意义在于 将促进对新型 RyR2 磷酸化位点和 SPEG 生理重要性的理解 激酶活性。这些知识有可能有助于确定新的治疗靶点 治疗心力衰竭等心血管疾病。

项目成果

期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

Hannah Moore Campbell其他文献

Hannah Moore Campbell的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

相似海外基金

Collaborative Research: REU Site: Earth and Planetary Science and Astrophysics REU at the American Museum of Natural History in Collaboration with the City University of New York
合作研究:REU 地点:地球与行星科学和天体物理学 REU 与纽约市立大学合作,位于美国自然历史博物馆
  • 批准号:
    2348998
  • 财政年份:
    2025
  • 资助金额:
    $ 4.21万
  • 项目类别:
    Standard Grant
Collaborative Research: REU Site: Earth and Planetary Science and Astrophysics REU at the American Museum of Natural History in Collaboration with the City University of New York
合作研究:REU 地点:地球与行星科学和天体物理学 REU 与纽约市立大学合作,位于美国自然历史博物馆
  • 批准号:
    2348999
  • 财政年份:
    2025
  • 资助金额:
    $ 4.21万
  • 项目类别:
    Standard Grant
Understanding Latin American Challenges in the 21st Century (LAC-EU)
了解拉丁美洲在 21 世纪面临的挑战 (LAC-EU)
  • 批准号:
    EP/Y034694/1
  • 财政年份:
    2024
  • 资助金额:
    $ 4.21万
  • 项目类别:
    Research Grant
Conference: North American High Order Methods Con (NAHOMCon)
会议:北美高阶方法大会 (NAHOMCon)
  • 批准号:
    2333724
  • 财政年份:
    2024
  • 资助金额:
    $ 4.21万
  • 项目类别:
    Standard Grant
Collaborative Research: RUI: Continental-Scale Study of Jura-Cretaceous Basins and Melanges along the Backbone of the North American Cordillera-A Test of Mesozoic Subduction Models
合作研究:RUI:北美科迪勒拉山脊沿线汝拉-白垩纪盆地和混杂岩的大陆尺度研究——中生代俯冲模型的检验
  • 批准号:
    2346565
  • 财政年份:
    2024
  • 资助金额:
    $ 4.21万
  • 项目类别:
    Standard Grant
REU Site: Research Experiences for American Leadership of Industry with Zero Emissions by 2050 (REALIZE-2050)
REU 网站:2050 年美国零排放工业领先地位的研究经验 (REALIZE-2050)
  • 批准号:
    2349580
  • 财政年份:
    2024
  • 资助金额:
    $ 4.21万
  • 项目类别:
    Standard Grant
Collaborative Research: RUI: Continental-Scale Study of Jura-Cretaceous Basins and Melanges along the Backbone of the North American Cordillera-A Test of Mesozoic Subduction Models
合作研究:RUI:北美科迪勒拉山脊沿线汝拉-白垩纪盆地和混杂岩的大陆尺度研究——中生代俯冲模型的检验
  • 批准号:
    2346564
  • 财政年份:
    2024
  • 资助金额:
    $ 4.21万
  • 项目类别:
    Standard Grant
Conference: Latin American School of Algebraic Geometry
会议:拉丁美洲代数几何学院
  • 批准号:
    2401164
  • 财政年份:
    2024
  • 资助金额:
    $ 4.21万
  • 项目类别:
    Standard Grant
Collaborative Research: Ionospheric Density Response to American Solar Eclipses Using Coordinated Radio Observations with Modeling Support
合作研究:利用协调射电观测和建模支持对美国日食的电离层密度响应
  • 批准号:
    2412294
  • 财政年份:
    2024
  • 资助金额:
    $ 4.21万
  • 项目类别:
    Standard Grant
Conference: Doctoral Consortium at Student Research Workshop at the Annual Conference of the North American Chapter of the Association for Computational Linguistics (NAACL)
会议:计算语言学协会 (NAACL) 北美分会年会学生研究研讨会上的博士联盟
  • 批准号:
    2415059
  • 财政年份:
    2024
  • 资助金额:
    $ 4.21万
  • 项目类别:
    Standard Grant
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了