Post-transcriptional mechanisms of gene regulation in cardiac cell growth and development

心肌细胞生长发育中基因调控的转录后机制

基本信息

批准号：
10642893
负责人：
Auinash Kalsotra
金额：
$ 38.56万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-04-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10642893
关键词：
Adult Affect Antisense Oligonucleotides Antisense RNA Binding Proteins Binding Sites Biological Assay CRISPR/Cas technology Cardiac Cardiac Myocytes Cause of Death Cell Nucleus Compensation Cues Cytoplasm Development Disease Funding Gene Expression Gene Expression Regulation Gene Silencing Genetic Goals Growth Growth and Development function Heart Heart Diseases Heart Hypertrophy Heart failure Hypertrophy Knock-out Knockout Mice Laboratories Length Mediating Messenger RNA Molecular Mus Muscle Muscle Cells Myocardial Myocardial Infarction Myocardium Neonatal Nuclear Outcome Pathologic Performance Physiological Poly A Poly(A) Tail Poly(A)-Binding Proteins Polyadenylation Polyribosomes Post-Transcriptional Regulation Proliferating Protein Biosynthesis RNA RNA Splicing Regulation Regulatory Pathway Risk Factors Role Shapes Signal Transduction Skeletal Muscle Stimulus Stress Tamoxifen Testing Tetracyclines Therapeutic Tissues Titrations Trans-Activators Transcript Transgenic Mice Translating Translations United States Work cardiogenesis cell growth experimental study fetal genome editing heart function hemodynamics in vivo insight mRNA Export mouse model overexpression polysome profiling postnatal postnatal development posttranscriptional premature programs protein expression response single molecule superresolution microscopy transcription termination transcriptome transcriptome sequencing translational model translatome

项目摘要

I. ABSTRACT Heart disease remains the leading cause of death in the United States. Despite what we know about the risk factors associated with heart disease, the molecular mechanisms are still largely unknown. The healthy adult heart is unique from other tissues in that the rate of protein synthesis is dramatically lower than in most tissues and lower than the developing heart. However, during cardiac hypertrophy, translation rates increase. This suggests a tissue-specific mechanism for regulating translation rates in the mammalian heart. Our lab has identified a mechanism by which total protein synthesis in cardiomyocytes is decreased during development through shortening of poly(A) tails, leading to a decrease in polysome formation through the closed-loop model of translation. This regulation is reversed during both physiologic and pathologic hypertrophy when the translation needs of cardiomyocytes are increased. Also, we have discovered that the nuclear poly(A) binding protein (PABPN1) is post-transcriptionally silenced in mammalian adult cardiac and skeletal muscle but it becomes re-expressed in pathologic cardiac hypertrophy. PABPN1 is a regulator of alternative polyadenylation (APA) and poly(A) tail length, both of which can influence the translation of transcripts. Our central hypothesis is that PABPN1 is dynamically regulated in cardiac myocytes to tune translation rates and suite growth needs through a polyadenylation dependent mechanism. The objective of this proposal is to elucidate the exact function(s) of PABPN1 in cardiac development and growth and identify how PABPN1 is regulated during these conditions. Aims 1 and 2 will use conditional PABPN1-knockout and overexpressing mice to determine the physiologic roles of PABPN1 in cardiac development and hypertrophy while defining the molecular basis of PABPN1 activity and its role in determining cardiac-specific gene expression programs. In Aim 3, we will use super-resolution microscopy, CRISPR-Cas9 mediated genome editing, and RNA antisense-oligo pulldown approaches to identify the regulatory mechanism(s) and factors that post-transcriptionally silence PABPN1 during cardiac development.

I.摘要心脏病仍然是美国死亡的主要原因。尽管我们知道与心脏病相关的危险因素，分子机制仍然在很大程度上未知。健康成人心脏与其他组织是独一无二的，因为蛋白质合成的速率大大低于大多数组织，低于发育中的心脏。但是，在心脏肥大期间，翻译率会增加。这表明一种组织特异性的机制，用于调节哺乳动物心脏的翻译率。我们的实验室有确定了一种机制，通过该机制，在发育过程中，心肌细胞中总蛋白质合成降低通过缩短聚（a）尾巴，从而通过闭环模型减少多元体组形成翻译。当生理和病理肥大期间，这种调节在心肌细胞的翻译需求增加。另外，我们发现核电（a）结合蛋白质（PABPN1）在哺乳动物的成人心脏和骨骼肌中在转录后沉默，但在病理心脏肥大中重新表达。 PABPN1是替代聚腺苷酸化的调节剂（APA）和poly（a）尾巴长度，两者都会影响转录本的翻译。我们的中心假设是在心肌细胞中动态调节PABPN1以调整翻译率和套件的增长需求通过依赖于聚腺苷酸化的机制。该提议的目的是阐明 PABPN1在心脏发展和生长中的功能，并确定在这些过程中如何调节PABPN1 状况。目标1和2将使用条件PABPN1敲除和过表达的小鼠来确定 PABPN1在心脏发育和肥大中的生理作用，同时定义分子基础 PABPN1活性及其在确定心脏特异性基因表达程序中的作用。在AIM 3中，我们将使用超分辨率显微镜，CRISPR-CAS9介导的基因组编辑和RNA反义 - oligo pollldown 确定调节机制和转录后沉默PABPN1的因素的方法在心脏发展期间。

项目成果

期刊论文数量（15）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Deregulation of RNA Metabolism in Microsatellite Expansion Diseases.

微卫星扩增疾病中 RNA 代谢失调。

DOI：
10.1007/978-3-319-89689-2_8
发表时间：
2018
期刊：
Advances in neurobiology
影响因子：
0
作者：
Misra,Chaitali;Lin,Feikai;Kalsotra,Auinash
通讯作者：
Kalsotra,Auinash

Impaired Mitochondrial Energy Production Causes Light-Induced Photoreceptor Degeneration Independent of Oxidative Stress.

DOI：
10.1371/journal.pbio.1002197
发表时间：
2015-07
期刊：
PLoS biology
影响因子：
9.8
作者：
Jaiswal M;Haelterman NA;Sandoval H;Xiong B;Donti T;Kalsotra A;Yamamoto S;Cooper TA;Graham BH;Bellen HJ
通讯作者：
Bellen HJ

RNA modifications and structures cooperate to guide RNA-protein interactions.

DOI：
10.1038/nrm.2016.163
发表时间：
2017-03
期刊：
Nature reviews. Molecular cell biology
影响因子：
0
作者：
Lewis CJ;Pan T;Kalsotra A
通讯作者：
Kalsotra A

Alternative splicing rewires Hippo signaling pathway in hepatocytes to promote liver regeneration.

DOI：
10.1038/s41594-018-0129-2
发表时间：
2018-10
期刊：
Nature structural & molecular biology
影响因子：
16.8
作者：
Bangru S;Arif W;Seimetz J;Bhate A;Chen J;Rashan EH;Carstens RP;Anakk S;Kalsotra A
通讯作者：
Kalsotra A

Complementary Oligonucleotide Conjugated Multicolor Carbon Dots for Intracellular Recognition of Biological Events.

DOI：
10.1021/acsami.0c02463
发表时间：
2020-04-08
期刊：
ACS applied materials & interfaces
影响因子：
9.5
作者：
Srivastava I;Misra SK;Bangru S;Boateng KA;Soares JANT;Schwartz-Duval AS;Kalsotra A;Pan D
通讯作者：
Pan D