Establishing and reversing the functional consequences of Titin truncation mutations
建立并逆转肌联蛋白截断突变的功能后果
基本信息
- 批准号:10640157
- 负责人:
- 金额:$ 52.91万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2022
- 资助国家:美国
- 起止时间:2022-07-01 至 2026-06-30
- 项目状态:未结题
- 来源:
- 关键词:3-DimensionalAdrenergic AgentsAgar Gel ElectrophoresisAlternative SplicingBase PairingBehaviorBiological AssayBiologyBiomimeticsCRISPR-mediated transcriptional activationCRISPR/Cas technologyCardiacCardiac MyocytesCardiomyopathiesCellsClustered Regularly Interspaced Short Palindromic RepeatsCommunitiesComputing MethodologiesDNADataDependenceDevelopmentDiagnosisDilated CardiomyopathyDiseaseDistalDoseExtracellular MatrixFibroblastsFunctional disorderFutureGenesGeneticGoalsGuide RNAHeartHeart failureHeterozygoteHumanIndividualKineticsKnowledgeLearningLengthLesionLinkLocationMeasuresMechanicsMediatingMethodsModelingMolecularMusMutationMyocardial ContractionMyocardiumNonmuscle Myosin Type IIAOrganellesPathogenesisPathogenicityPathologicPathway interactionsPatientsPhenotypePhysiciansPhysiologicalPrognosisProtein IsoformsProteinsPublicationsRNA BindingReagentRegulationRegulatory ElementResearchResourcesRiskSarcomeresSeriesSignal TransductionSiteStressStructureSturnus vulgarisSudden DeathTechnologyTherapeuticTissue EngineeringTissue ModelTissue-Specific Gene ExpressionTranscription CoactivatorVariantWestern BlottingWorkadverse outcomebiological adaptation to stressbiomarker developmentbiomarker identificationcardiac tissue engineeringcohortconfocal imagingconnectindisease prognosisdosagegenome editingheart cellheart dimension/sizeheart functionhigh riskhuman modelimprovedinduced pluripotent stem cellinduced pluripotent stem cell derived cardiomyocytesinherited cardiomyopathyinsightmechanotransductionmutation carrierpreventpromoterproteostasisresponsescaffoldsudden cardiac deaththerapeutic developmenttherapeutic targettranscriptomics
项目摘要
PROJECT SUMMARY/ABSTRACT
Cardiomyopathies occur in ~1:200 individuals and are commonly caused by inheritance of variants in genes that
encode proteins that regulate the sarcomere, the force-producing organelle of heart cells. Due to an incomplete
understanding of variant pathogenicity and cardiomyopathy pathogenesis, physicians are currently limited in
their ability to provide diagnoses, prognoses, and therapeutic options for cardiomyopathy patients. Variants in
the TTN gene, which encodes the sarcomere protein titin, are the most frequently identified genetic lesion in
dilated cardiomyopathy (DCM), which is characterized by heart chamber dilation, reduced contractile function,
risk of sudden death, and progressive heart failure. The most frequent type of TTN variant identified in DCM is
a truncation mutation that would be predicted to shorten TTN protein length and to reduce TTN protein quantities.
Significantly, truncation variants localized to distal TTN structural domains are more pathogenic than those
localized to proximal structural domains, but the mechanistic basis for this relationship is uncertain. It remains
incompletely understood how TTN truncation variants cause DCM generally, which is compounded by our lack
of understanding of the ‘length dependence’ of TTN variant pathogenicity. These knowledge gaps limit disease
prognostication, biomarker identification, and therapeutic development for DCM patients. The central goal of
our study is to define how disruptions in TTN length and dosage by TTN variants cause DCM, and exploit this
knowledge to develop DCM therapeutics for TTN variant carriers. We hypothesize that healthy cardiac
contractile function and structure depends on the regulation of TTN length and dosage, and that varying
pathogenicity of TTN truncation can be explained by distinct structural and functional consequences associated
with the specific site of truncation. In Aim 1, we will determine the functional consequences of TTN truncations
across structural domains by harnessing 3-dimensional heart tissue models composed of human cardiomyocytes
differentiated from induced pluripotent stem cells in which variants have been introduced by CRISPR-mediated
genome editing. We will interrogate these models for tissue mechanical phenotypes (such as passive tension
and Frank-Starling behavior), TTN protein length and levels (using specialized methods), proteostasis stress
pathway responses (using immunoblotting), and mechanotransduction signaling and alternative splicing (using
expression analysis and transcriptomics, respectively). In Aim 2, we will restore TTN protein levels using the
recently developed method of CRISPR activation applied to DCM engineered heart tissue models for both
evaluating the function of TTN isoforms generally and as a DCM proof-of-concept therapeutic. Through these
Aims, we will gain critical new insights into the pathophysiology of DCM-associated TTN truncation variants,
uncover features to explain the variable pathogenicity identified in DCM patients, and develop a therapeutic to
target TTN directly. We anticipate this new knowledge will improve physicians’ capacity to diagnose, prognose,
and treat patients with DCM due to TTN variants.
项目总结/摘要
心肌病发生在约1:200的个体中,通常由遗传基因变异引起,
编码调节肌节的蛋白质,肌节是心脏细胞产生力量的细胞器。由于不完整的
了解变异致病性和心肌病发病机制,医生目前在
他们为心肌病患者提供诊断、诊断和治疗选择的能力。中的变体
编码肌节蛋白肌联蛋白的TTN基因是最常见的遗传性病变,
扩张型心肌病(DCM),其特征在于心腔扩张,收缩功能降低,
猝死和进行性心力衰竭的风险。DCM中最常见的TTN变异类型是
一个截断突变,预计将缩短TTN蛋白长度和减少TTN蛋白量。
值得注意的是,定位于远端TTN结构域的截短变体比那些截短变体更具致病性。
定位于近端结构域,但这种关系的机制基础是不确定的。它仍然是
不完全理解TTN截短变体通常如何导致DCM,这与我们缺乏
了解TTN变异致病性的“长度依赖性”。这些知识差距限制了疾病
扩张型心肌病患者的诊断、生物标志物鉴定和治疗开发。的中心目标
我们的研究是为了确定TTN变异体如何破坏TTN长度和剂量导致DCM,并利用这一点。
开发TTN变异携带者DCM治疗方法的知识。我们假设健康的心脏
收缩功能和结构取决于TTN长度和剂量的调节,
TTN截短的致病性可以通过与TTN截短相关的不同结构和功能后果来解释。
与特定的截短位点。在目标1中,我们将确定TTN截短的功能后果
通过利用由人类心肌细胞组成的三维心脏组织模型,
从诱导的多能干细胞分化,其中变体已经通过CRISPR介导的CRISPR介导的免疫调节被引入。
基因组编辑我们将询问这些模型的组织力学表型(如被动张力
和Frank-Starling行为),TTN蛋白长度和水平(使用专门的方法),蛋白质稳态应激
途径反应(使用免疫印迹),以及机械转导信号和可变剪接(使用
表达分析和转录组学)。在目标2中,我们将使用以下方法恢复TTN蛋白水平:
最近开发的CRISPR激活方法应用于DCM工程化心脏组织模型,
一般评价TTN同种型的功能和作为DCM概念验证治疗剂的功能。通过这些
目的是,我们将获得对DCM相关TTN截短变体的病理生理学的重要新见解,
揭示特征以解释DCM患者中鉴定的可变致病性,并开发治疗方法,
直接瞄准TTN我们预计这一新知识将提高医生的诊断能力,
并治疗因TTN变异而患DCM的患者。
项目成果
期刊论文数量(1)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
TTN truncation variants produce sarcomere-integrating proteins of uncertain functional significance.
- DOI:10.1172/jci175206
- 发表时间:2024-01-16
- 期刊:
- 影响因子:15.9
- 作者:Hinson, J. Travis;Campbell, Stuart G.
- 通讯作者:Campbell, Stuart G.
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STUART G CAMPBELL其他文献
STUART G CAMPBELL的其他文献
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{{ truncateString('STUART G CAMPBELL', 18)}}的其他基金
Establishing and reversing the functional consequences of Titin truncation mutations
建立并逆转肌联蛋白截断突变的功能后果
- 批准号:
10510011 - 财政年份:2022
- 资助金额:
$ 52.91万 - 项目类别:
Computer modeling of myosin binding protein C and its effects on cardiac contraction
肌球蛋白结合蛋白 C 的计算机建模及其对心脏收缩的影响
- 批准号:
10371076 - 财政年份:2019
- 资助金额:
$ 52.91万 - 项目类别:
Computer modeling of myosin binding protein C and its effects on cardiac contraction
肌球蛋白结合蛋白 C 的计算机建模及其对心脏收缩的影响
- 批准号:
9903433 - 财政年份:2019
- 资助金额:
$ 52.91万 - 项目类别:
Revealing Pathomechanisms of Mutant TPM1 Through a Hybrid Computational-Experimental Approach
通过混合计算-实验方法揭示突变 TPM1 的病理机制
- 批准号:
10358783 - 财政年份:2017
- 资助金额:
$ 52.91万 - 项目类别:
Revealing Pathomechanisms of Mutant TPM1 Through a Hybrid Computational-Experimental Approach
通过混合计算-实验方法揭示突变 TPM1 的病理机制
- 批准号:
9398261 - 财政年份:2017
- 资助金额:
$ 52.91万 - 项目类别:
Revealing Pathomechanisms of Mutant TPM1 Through a Hybrid Computational-Experimental Approach
通过混合计算-实验方法揭示突变 TPM1 的病理机制
- 批准号:
9983135 - 财政年份:2017
- 资助金额:
$ 52.91万 - 项目类别:
Revealing Pathomechanisms of Mutant TPM1 Through a Hybrid Computational-Experimental Approach
通过混合计算-实验方法揭示突变 TPM1 的病理机制
- 批准号:
10221767 - 财政年份:2017
- 资助金额:
$ 52.91万 - 项目类别:
Computational Pipeline for Identification of Disease-Causing Variants in Genes of the Cardiac Sarcomere
用于鉴定心脏肌节基因致病变异的计算流程
- 批准号:
10736459 - 财政年份:2017
- 资助金额:
$ 52.91万 - 项目类别:
Engineered Tissue for Biomechanical Phenotyping of Cardiomyopathy Patients
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- 批准号:
8974854 - 财政年份:2014
- 资助金额:
$ 52.91万 - 项目类别:
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