Decoding the electric face: development and channelopathy-induced birth defects

解码电脸：发育和通道病引起的出生缺陷

• 批准号: 8736077
• 负责人: DANY SPENCER ADAMS
• 金额: $ 32.18万
• 依托单位: TUFTS UNIVERSITY MEDFORD
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-15 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8736077
• 关键词: Address; Affect; Age; Amphibia; Anatomy; Angelman Syndrome; Area; Arrhythmia; Atlases; Biomedical Engineering; Biophysics; Birth; Cell membrane; Cells; Complement; Complex; Computer Simulation; Congenital Abnormality; Coupled; Data; Data Set; Defect; Development; Dyes; Eating; Embryo; Epigenetic Process; Etiology; Event; Eye; Eye Development; Face; Feedback; Fluorescent Dyes; Foundations; Future; Gap Junctions; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Processes; Growth; Head; Human; Hybrids; In Situ; Individual; Ion Channel; Ion Pumps; Ions; Knowledge; Life; Light; Link; Location; Mediating; Membrane; Membrane Potentials; Methods; Mining; Modeling; Molecular; Molecular Genetics; Morphogenesis; Morphology; Mutation; Natural regeneration; Neurologic; Organ; Pathway interactions; Patients; Pattern; Physiological; Primordium; Process; Property; Rana; Reading; Reagent; Regenerative Medicine; Regulation; Relative (related person); Reporter; Rest; Rewards; Role; Shapes; Signal Transduction; Source; Structure; Study models; Syndrome; Techniques; Testing; Time; Tissues; Work; Writing; Xenopus; base; bioelectricity; biophysical properties; cell behavior; chemical genetics; craniofacial; disease-causing mutation; electric field; in vivo; limb regeneration; loss of function; mathematical model; minimally invasive; novel; novel strategies; optogenetics; prevent; public health relevance; regional difference; relating to nervous system; repaired; self assembly; self organization; tool; tumorigenesis; voltage

DESCRIPTION (provided by applicant): Craniofacial development in tractable models such as the frog Xenopus, has been a popular and highly rewarding model for studies of the cell activities that build complex structures. Much information has been gathered about the chemical and genetic processes that underlie craniofacial patterning. In contrast to and complementing that work, our lab has discovered a layer of biophysical signaling: a regulated distribution of different cell membrane resting potentials that instruct cell behavior and large-scale morphogenesis. We have showed that eye development, limb regeneration, and tumorigenesis are all regulated by ion channel-mediated voltage gradients. We have adapted voltage-sensitive fluorescent dyes to observe these gradients and used molecular tools to modify them thereby regulating individual cell behaviors and reprogramming whole organ primordia. These techniques are distinct from classical methods of electric field application, and reveal not only the molecular-genetic sources of the gradients but also the epigenetic and transcriptional downstream steps through which biophysical properties regulate morphology. Recently, we uncovered a remarkable feature of early craniofacial development: the "electric face" - dynamic patterns of hyper- and depolarization in the embryonic frog face that precede, predict, and control the shape and location of facial structures. Perturbing these patterns, results in predictable changes in expression of key genes and changes in anatomy. This finding suggests a hypothesis for the heretofore-mysterious observation that several channelopathies (diseases caused by mutations in ion channel genes such as KCNJ2) cause not only neurological deficits and cardiac arrhythmias but also craniofacial defects. The fact that these channels participate in the establishment of the normal bioelectric regionalization of the embryonic face may explain why channels are essential for craniofacial patterning. Our project aims to: 1) characterize in detail the bioelectric properties of the early face relative to gene expression domains (a physiomics atlas merging transcriptional and biophysical data); 2) explore the emergence of the bioelectric face patterns by formulating a predictive mathematical model of self-organization within electrically-coupled cells; 3) reveal molecular details of how voltage gradients regulate specific downstream face-patterning gene expression domains; and 4) develop optogenetic techniques to read/write desired electrical patterns in living tissue to override incorrect membrane voltage patterns thus preventing defects. The resulting data will: serve as an essential foundation for future attempts to merge biophysical and transcriptional regulatory layers in order to explain complex development; establish a quantitative model to prescribe minimally invasive corrective manipulations of voltage-dependent patterning; and to chart a new approach toward exploiting guided self-assembly of bioelectrical patterns to address issues in regenerative medicine, etiology and treatment of birth defects, as well as produce new hybrid constructs via synthetic bioengineering.

描述（由申请人提供）：诸如青蛙爪蟾等可牵引模型中的颅面开发，是对构建复杂结构的细胞活动的研究的流行且高度奖励的模型。已经收集了有关颅面模式下的化学和遗传过程的许多信息。与这项工作相反并补充了这项工作，我们的实验室发现了一层生物物理信号传导：不同细胞膜静息电势的调节分布指导细胞行为和大规模形态发生。我们已经表明，眼睛发育，肢体再生和肿瘤发生都受离子通道介导的电压梯度调节。我们已经适应了对电压敏感的荧光染料，以观察这些梯度，并使用分子工具来修改它们，从而调节单个细胞行为并重新编程整个器官原始。这些技术与电场应用的经典方法不同，不仅揭示了梯度的分子遗传学源，还揭示了生物物理特性调节形态的表观遗传和转录下游步骤。最近，我们发现了早期颅面发育的显着特征：“电脸” - 胚胎青蛙面对面的超极化的动态模式，该模式是前面，预测和控制面部结构的形状和位置。扰动这些模式，导致关键基因表达的可预测变化和解剖学的变化。这一发现表明，迄今为止的观察结果是一种假设，即几种通道病（由ION通道基因（例如KCNJ2）引起的疾病）不仅会导致神经学缺陷和心律不齐，还导致颅面缺陷。这些渠道参与了胚胎面部正常生物电区域化的建立，这可能解释了为什么渠道对于颅面模式至关重要。我们的项目的目的是：1）详细介绍早期面相对于基因表达结构域的生物电特性（一种合并转录和生物物理数据的生理学图集）； 2）通过在电耦合细胞中制定预测的数学模型来探索生物电的面部模式的出现； 3）揭示了电压梯度如何调节特定的下游面部阴影基因表达结构域的分子细节； 4）开发光遗传技术来读/编写生物组织中所需的电气模式，以覆盖不正确的膜电压模式，从而防止缺陷。最终的数据将成为未来尝试合并生物物理和转录调节层以解释复杂发展的基础；建立一个定量模型，以规定对电压依赖性图案的微创矫正操作；并绘制一种新的方法来利用生物电模式的指导自组装，以解决再生医学，病因和治疗先天缺陷的问题，并通过合成生物工程生产新的混合构建体。