工程组织的几何形状和 Plakophilin-2 可调节人类 iPSC 衍生心肌细胞的电生理学。

IF 6.6 3区 医学 Q1 ENGINEERING, BIOMEDICAL
APL Bioengineering Pub Date : 2024-03-11 eCollection Date: 2024-03-01 DOI:10.1063/5.0160677
Daniel W Simmons, Ganesh Malayath, David R Schuftan, Jingxuan Guo, Kasoorelope Oguntuyo, Ghiska Ramahdita, Yuwen Sun, Samuel D Jordan, Mary K Munsell, Brennan Kandalaft, Missy Pear, Stacey L Rentschler, Nathaniel Huebsch
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引用次数: 0

摘要

人造心脏组织是为了在比二维单层培养更接近体内心肌的环境中研究心脏生物学和疾病。以前发表的研究表明,几何各向异性的微环境对诱导未成熟心肌细胞产生 "类似于体内 "的生理机能至关重要。我们假设,工程组织内心肌细胞的排列和预应力程度受组织几何形状的调节,并随之推动电生理的发展。因此,我们研究了组织几何形状对由人类诱导多能干细胞(iPSCs)设计的微型心肌阵列(μHM)电生理学的影响。拉长的组织几何形状引起了心肌细胞形状和电生理学的变化,从而产生适应性,增加了每个收缩周期的钙摄入量。令人震惊的是,药理学研究显示,钠通道功能需要预应力和/或细胞排列的阈值,而 L 型钙通道和快速整流钾通道对这些变化基本不敏感。同时,组织伸长上调了钠通道(NaV1.5)和缝隙连接(Connexin 43,Cx43)蛋白的表达。基于这些观察结果,我们利用拉长的μHM来研究Plakophilin 2(PKP2)功能缺失突变的影响。在μHM中,PKP2基因敲除心肌细胞的细胞形态与在同源对照组中观察到的相似。然而,PKP2-/-组织的传导速度较低,且没有功能性钠流。PKP2基因敲除的μHM表现出钠离子通道的几何关联上调,而不是Cx43,这表明翻译后机制,包括离子通道-间隙连接沟通的缺乏,可能是在携带这种基因缺陷的组织中观察到的较低传导速度的原因。总之,这些观察结果表明,简单、可扩展的微组织系统可以提供必要的生理压力,诱导 iPS-CM 的电重塑,从而实现对疾病相关基因组变异的电生理后果的研究。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Engineered tissue geometry and Plakophilin-2 regulate electrophysiology of human iPSC-derived cardiomyocytes.

Engineered heart tissues have been created to study cardiac biology and disease in a setting that more closely mimics in vivo heart muscle than 2D monolayer culture. Previously published studies suggest that geometrically anisotropic micro-environments are crucial for inducing "in vivo like" physiology from immature cardiomyocytes. We hypothesized that the degree of cardiomyocyte alignment and prestress within engineered tissues is regulated by tissue geometry and, subsequently, drives electrophysiological development. Thus, we studied the effects of tissue geometry on electrophysiology of micro-heart muscle arrays (μHM) engineered from human induced pluripotent stem cells (iPSCs). Elongated tissue geometries elicited cardiomyocyte shape and electrophysiology changes led to adaptations that yielded increased calcium intake during each contraction cycle. Strikingly, pharmacologic studies revealed that a threshold of prestress and/or cellular alignment is required for sodium channel function, whereas L-type calcium and rapidly rectifying potassium channels were largely insensitive to these changes. Concurrently, tissue elongation upregulated sodium channel (NaV1.5) and gap junction (Connexin 43, Cx43) protein expression. Based on these observations, we leveraged elongated μHM to study the impact of loss-of-function mutation in Plakophilin 2 (PKP2), a desmosome protein implicated in arrhythmogenic disease. Within μHM, PKP2 knockout cardiomyocytes had cellular morphology similar to what was observed in isogenic controls. However, PKP2-/- tissues exhibited lower conduction velocity and no functional sodium current. PKP2 knockout μHM exhibited geometrically linked upregulation of sodium channel but not Cx43, suggesting that post-translational mechanisms, including a lack of ion channel-gap junction communication, may underlie the lower conduction velocity observed in tissues harboring this genetic defect. Altogether, these observations demonstrate that simple, scalable micro-tissue systems can provide the physiologic stresses necessary to induce electrical remodeling of iPS-CM to enable studies on the electrophysiologic consequences of disease-associated genomic variants.

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来源期刊
APL Bioengineering
APL Bioengineering ENGINEERING, BIOMEDICAL-
CiteScore
9.30
自引率
6.70%
发文量
39
审稿时长
19 weeks
期刊介绍: APL Bioengineering is devoted to research at the intersection of biology, physics, and engineering. The journal publishes high-impact manuscripts specific to the understanding and advancement of physics and engineering of biological systems. APL Bioengineering is the new home for the bioengineering and biomedical research communities. APL Bioengineering publishes original research articles, reviews, and perspectives. Topical coverage includes: -Biofabrication and Bioprinting -Biomedical Materials, Sensors, and Imaging -Engineered Living Systems -Cell and Tissue Engineering -Regenerative Medicine -Molecular, Cell, and Tissue Biomechanics -Systems Biology and Computational Biology
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