Investigating the Early Embryonic Murine Heart Using Optical Coherence Tomography

使用光学相干断层扫描研究早期胚胎小鼠心脏

• 批准号: 7908895
• 负责人: ANDREW Martin ROLLINS
• 金额: $ 75.81万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-05 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7908895
• 关键词: Address; Algorithms; Anatomy; Animal Model; Arts; Biological; Birds; Blood flow; Cardiac; Cardiology; Complex; Congenital Heart Defects; Cytoskeletal Modeling; Data; Defect; Development; Developmental Biology; Embryo; Embryo Culture Techniques; Embryonic Development; Embryonic Heart; Emerging Technologies; Engineering; Financial compensation; Frequencies; Gene Expression; Gene Proteins; Genes; Genome Mappings; Goals; Heart; Human; Image; Image Analysis; Imagery; Imaging Device; Imaging technology; In Vitro; Incubated; Investigation; Lasers; Life; Maps; Measurement; Medical; Methods; Microscopic; Microscopy; Modeling; Molecular; Morphogenesis; Morphology; Motion; Mus; Mutant Strains Mice; Noise; Optical Coherence Tomography; Optics; Pattern; Phase; Pregnancy; Process; Research; Resolution; Scanning; Shapes; Side; Software Tools; Speed; Staging; Stress; Structure; System; Techniques; Technology; Testing; Three-Dimensional Image; Time; Tissue Sample; Tissues; Transgenic Mice; Ultrasonography; Validation; Zebrafish; base; cardiogenesis; embryo culture; heart function; hemodynamics; imaging Segmentation; improved; in utero; interest; next generation; novel; optical imaging; particle; placental mammal; protein expression; public health relevance; reconstruction; research study; shear stress; statistics; success; tool

DESCRIPTION (provided by applicant): The embryonic murine heart is a very important model of human heart development due to its four- chambered structure similar to the human heart, a short gestation period and a completely mapped genome which is easy to alter and manipulate. Lack of an appropriate imaging technology has previously hindered progress in uncovering the normal/abnormal mechanisms that govern early heart development in mice. The multi-disciplinary project described in this five-year proposal will develop, investigate and validate imaging and analysis tools to rapidly and thoroughly investigate the living embryonic murine heart at a level of spatial and temporal resolution previously not possible. These tools are based on the emerging technology of optical coherence tomography (OCT), which is capable of micrometer-scale resolution imaging of small biological tissue samples non-destructively and in real time. Compared to avian and zebrafish models, the murine embryo is far more challenging to culture in vitro in critical early looping stages and thus presents unique imaging challenges. We will develop methods and technology to culture mouse embryos that enable normal development and facilitate optical imaging. The significant potential of OCT for embryonic imaging is clear. We will develop the technology to fully realize this potential by improving imaging speed, resolution and scanning technology to enable complete characterization of the morphology and contractile and hemodynamic function of the early embryonic murine heart. Data interpretation is a key to the success of investigations using imaging. We will develop a suite of advanced tools for 2D/3D image preprocessing, analysis and visualization that will facilitate examination and comparison of the acquired image data. These tools, including noise reduction, registration, intensity inhomogeneity compensation, visualization using opacity optimization, semi-automatic and supervised 3D image segmentation, and tissue displacement and blood flow mapping, will facilitate new observations and discovery from these unique data. A set of baseline experiments will establish the usefulness of our OCT technology and image analysis tools during the stages of most dramatic cardiac morphogenesis. We will compare functional measurements obtained in vitro from OCT with those obtained in utero using micro ultrasound. We will investigate the effect of altered hemodynamics in NMHC-IIB transgenic ( and -/-) mice on heart looping. Finally, we will correlate gene expression with functional measurements made from OCT data. The end product of this effort will be a novel set of imaging tools based on OCT, partnered with improved embryo culturing technology, which have been optimized and validated for investigating the developing murine heart. The applicability of the developed technology will not only extend to other models of heart defects, but also to many other fields of developmental biology. The hypothesis tested in the validation aim will address critical open questions regarding the earliest functional changes leading to congenital heart defects. PUBLIC HEALTH RELEVANCE: Early medical treatments of congenital heart defects can only be developed if we understand the origins of the defects while the heart is first developing. The lack of an ideal way to image the structure and function of these tiny, almost microscopic hearts has limited our ability answer these questions. OCT imaging of embryonic mouse heart can fill this need and we will develop and validate the technology and methods to reach this potential.

描述（由申请人提供）：胚胎鼠心脏是人类心脏发育的非常重要的模型，因为它的四个腔室结构类似于人类的心脏，妊娠期短和完全映射的基因组，易于改变和操纵。缺乏适当的成像技术以前已经阻碍了揭示了控制小鼠早期心脏发育的正常/异常机制。这项五年提案中描述的多学科项目将开发，调查和验证成像和分析工具，以快速，彻底地研究以前无法实现的空间和时间分辨率水平的生活胚胎鼠心脏。这些工具基于光学相干断层扫描（OCT）的新兴技术，该技术能够实时和实时对小型生物组织样品进行微尺度的分辨率成像。与鸟类和斑马鱼模型相比，在关键的早期循环阶段，鼠类胚胎对体外的培养更具挑战性，因此提出了独特的成像挑战。我们将开发用于培养小鼠胚胎的方法和技术，从而使正常发育并促进光学成像。 OCT对胚胎成像的显着潜力很明显。我们将通过提高成像速度，分辨率和扫描技术来制定技术以充分实现这一潜力，从而使早期胚胎鼠心脏的形态和收缩性和血流动力学功能完全表征。数据解释是使用成像成功进行调查成功的关键。我们将开发一套高级工具，用于2D/3D图像预处理，分析和可视化，以促进对获得的图像数据的检查和比较。这些工具，包括降低噪声，注册，强度不均匀性补偿，使用不透明度优化的可视化，半自动和监督的3D图像分割以及组织位移和血流图映射，将促进这些独特数据的新观察和发现。一组基线实验将在大多数戏剧性心脏形态发生的阶段确定我们的OCT技术和图像分析工具的有用性。我们将比较从OCT中获得的功能测量与使用微观超声在子宫内获得的功能测量值。我们将研究NMHC-IIB转基因（和 - / - ）小鼠对心脏循环的血液动力学改变的影响。最后，我们将将基因表达与OCT数据制成的功能测量相关联。这项工作的最终产品将是基于OCT的新型成像工具，该工具与改进的胚胎培养技术合作，已对其进行优化和验证，以调查发育中的鼠心脏。开发技术的适用性不仅将扩展到其他心脏缺陷模型，还将扩展到许多其他发育生物学领域。在验证目标中检验的假设将解决有关导致先天性心脏缺陷的最早功能变化的关键开放问题。公共卫生相关性：只有在心脏首次发展时缺陷的起源时，才能开发出先天性心脏缺陷的早期药物治疗。缺乏对这些微小，几乎微观心脏的结构和功能的理想方法的理想方法，因此限制了我们的能力回答这些问题。胚胎小鼠心脏的OCT成像可以满足这一需求，我们将开发和验证达到这种潜力的技术和方法。