VIP-induced gene expression in colonic smooth muscle cells

VIP诱导结肠平滑肌细胞基因表达

• 批准号: 7275343
• 负责人: SUSHIL K SARNA
• 金额: $ 30.78万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-15 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7275343
• 关键词: Acetylcholine; Action Potentials; Adenosine Triphosphate; Adenylate Cyclase; Animals; Anti-Inflammatory Agents; Anti-inflammatory; Applications Grants; Bathing; Binding; Calcium; Cell physiology; Cells; Contracts; Coupling; Cyclic AMP; Cyclic AMP Response Element; Cyclic AMP-Responsive DNA-Binding Protein; Data; Differentiation and Growth; Disease; Elements; Enteral; Enteric Nervous System; Epigenetic Process; Equilibrium; Gene Expression; Gene Targeting; Genes; Genomics; Health; Homeostasis; Human; Immune system; Inflammation; Inflammatory; L-Type Calcium Channels; MAP Kinase Gene; Mediating; Mediator of activation protein; Molecular; Motor Neurons; Mucositis; Muscle; Muscle function; Muscle relaxation phase; Myenteric Plexus; Neural Conduction; Neuronal Injury; Neurons; Neuropeptides; Neurotransmitters; Nitric Oxide; Peptides; Personal Satisfaction; Phosphorylation; Play; Protein Binding; Regulation; Reporting; Research; Response Elements; Rest; Role; Signal Pathway; Signal Transduction; Smooth Muscle Myocytes; Sodium Channel; Substance P; Tetrodotoxin; Thinking; Vasoactive Intestinal Peptide; activating transcription factor; base; calmodulin-dependent protein kinase II; cytokine; gastrointestinal; inhibitory neuron; insight; non-genomic; novel; pituitary adenylate cyclase activating polypeptide; promoter; receptor; transcription factor

DESCRIPTION (provided by applicant): Vasoactive intestinal polypeptide (VIP) and pituitary adenylate cyclase-activating peptide (PACAP) are the inhibitory neurotransmitters in the gut wall. The mechanisms of the non-genomic effects of these neurotransmitters in smooth muscle relaxation have been investigated extensively over the last several decades. The genomic effects of these neurotransmitters on smooth muscle function are not known. VIP and PACAP, on binding to their receptors on smooth muscle cells, activate adenylate cyclase to produce cyclic 3'-5' adenosine monophosphate (cAMP) that mediates smooth muscle relaxation. However, cAMP is also a well known mediator of gene expression through the binding of transcription factor CRE binding protein (CREB) to the cAMP response element (CRE) on the promoters of its target genes. We have obtained substantial preliminary data that suggest an important novel function of the classic neurotransmitters VIP and PACAP to induce gene expression of the pore-forming alpha1C subunit of L-type Ca2+ channels in human colonic circular smooth muscle cells (HCCSMC). Ca2+ influx through these channels is an immediate early step in the signaling cascade for excitation-contraction coupling. Our preliminary data indicate that VIP/PACAP may also be anti-inflammatory neuropeptides that counter the initiation of the signaling cascade that activates the transcription factor NF-KB resulting in the suppression of cell contractility during inflammation. Based on these preliminary data our specific aims are to investigate: 1) VIP/PACAP-induced enhancement of alpha1C gene expression through cAMP/PKA signaling pathway in HCCSMC. 2) The cis- and trans-regulation of human ?1C promoter by VIP/PACAP-induced phosphorylation of the transcription factor CREB. 3) The interactions between cAMP/PKA, MAPK, PKC and CaMKII signaling pathways for the induction of alpha1C gene by VIP and PACAP. 4) The molecular and epigenetic mechanisms of the rnyo-protective role of VIP/PACAP in HCCSMC. This grant proposal presents a novel direction of research in the genomic regulation of smooth muscle function by two prominent and abundant neurotransmitters (VIP and PACAP) of the enteric inhibitory motor neurons. The findings will provide genomic and molecular insights into regulation of the expression of L-type calcium channels that play a critical role in excitation-contraction coupling in the normal state and during inflammation.

描述(申请人提供)：血管活性肠肽(VIP)和垂体腺苷环化酶激活肽(PACAP)是肠壁中的抑制性神经递质。在过去的几十年里，这些神经递质在平滑肌松弛中的非基因组效应的机制得到了广泛的研究。这些神经递质对平滑肌功能的基因组效应尚不清楚。VIP和PACAP与其受体结合后，激活腺苷环化酶，产生环状3‘-5’-腺苷一磷酸(CAMP)，介导平滑肌松弛。然而，cAMP也是通过转录因子CRE结合蛋白(CREB)与其靶基因启动子上的cAMP反应元件(CRE)结合来调节基因表达的。我们已经获得了大量的初步数据，表明经典神经递质VIP和PACAP在诱导人结肠环状平滑肌细胞(HCCSMC)中L型钙通道成孔α1C亚单位的基因表达方面具有重要的新功能。钙离子通过这些通道的内流是兴奋-收缩偶联的信号级联的第一步。我们的初步数据表明，VIP/PACAP也可能是抗炎神经肽，它可以对抗激活转录因子NF-KB的信号级联反应的启动，从而在炎症过程中抑制细胞的收缩。1)VIP/PACAP通过cAMP/PKA信号通路诱导HCCSMCα1C基因表达增强。2)VIP/PACAP诱导转录因子CREB的磷酸化对人β1C启动子的顺式和反式调节。3)VIP和PACAP诱导α1C基因表达的cAMP/PKA、MAPK、PKC和CaMKII信号通路之间的相互作用。4)VIP/PACAP对HCCSMC低温保护作用的分子和表观遗传学机制。这一资助方案为肠道抑制性运动神经元中两种重要和丰富的神经递质(VIP和PACAP)对平滑肌功能的基因组调控提供了一个新的研究方向。这些发现将为调控L型钙通道的表达提供基因组和分子方面的见解，该通道在正常状态和炎症过程中在兴奋-收缩偶联中发挥关键作用。