Fourier Light-Field Interrogation of Congenital Heart Disease in vivo

先天性心脏病(CHDS)是最常见的出生缺陷类型，影响全球约1%的人口。左心发育不全综合征(HLHS)是先天性心脏病婴儿最危险的形式和最常见的死亡原因。需要改进的成像技术来了解HLHS的机制，更好地实现早期诊断和干预。该项目将开发一种新的光场成像方法，能够在HLHS的活体动物模型中评估心脏的大小、结构和功能。利用这种方法获得的结果有望揭示导致HLHS的途径，并最终推动生物技术的发展，使心脏病患者受益。为了扩大这项工作的教育影响，研究人员将(1)通过校园内或在线资源优先招募妇女和少数族裔本科生和研究生参与这一项目，(2)通过组织亚特兰大生物光子学和高级光学显微镜国际学校，将研究和教育结合起来，(3)开发创新的教学方法，以加强以问题为基础的教育，提高公众的科学素养，以及(4)通过加强学校与实验室的互动和生物光子学夏令营，让未被充分代表的群体和退伍军人参与STEM。这些努力将影响下一代成像工程师和专业人员的培养，并促进不同学科的研究和教育的交叉融合。该项目的目标是开发和部署一个集成的光场功能屏幕平台，称为瞬时多色傅立叶光场显微镜(IFLFM)，用于同时、全光学地评估心脏形态和功能，以确定心脏病的病因基因。IFLFM系统将被应用于在体内监测这种疾病的新青蛙模型中HLHS的发展。该研究计划的制定有三个目标：(1)设计、构建和验证iFLFM，以便在组织和器官水平上实现快速、体积和同时的多颜色结构和动态成像，这将为拟议的在体询问胚胎蛙心脏的仪器平台奠定基础；(2)利用对照和Ets1敲除的青蛙胚胎验证iFLFM用于活体观察心脏的形态和功能，这将使心脏状况和缺陷的快照3D和多参数评估和记录成为可能，证实Ets1作为HLHS的致病基因的影响，并为拟议的其他候选基因的功能筛选准备基本的结构、分析和处理；以及(3)推进突变青蛙胚胎心脏功能筛选的iFLFM平台，以确定HLHS的因果基因，该平台将揭示和记录基因突变的解剖和功能变异和缺陷，并识别导致HLHS样表型的突变。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Congenital heart diseases (CHDs) are the most common type of birth defect and impact about 1% of the population worldwide. Hypoplastic left heart syndrome (HLHS) is the most dangerous form and the most common cause of death in infants with CHDs. Improved imaging technologies are needed to understand HLHS mechanisms and better achieve early diagnosis and intervention. This project will develop a novel light-field imaging method capable of assessing heart size, structure, and function in living animal models of HLHS. Results obtained leveraging this method are expected to reveal the pathways leading to HLHS and ultimately advance biotechnology for the benefit of patients with heart disease. To broaden the educational impacts of the work, the investigators will (1) prioritize and recruit women and minority undergraduate and graduate students to work on this project through on-campus or online resources, (2) integrate research and education by organizing an Atlanta international school on biophotonics and advanced optical microscopy, (3) develop innovative teaching and learning methods to enhance problem-based education and improve the public scientific literacy, and (4) engage underrepresented and veteran groups in STEM by enhancing school-lab interactions and through a summer camp on biophotonics. These efforts will impact the training of next-generation imaging engineers and professionals and promote the cross-fertilization of research and education from diverse disciplines.The goal of this project is to develop and deploy an integrative light-field functional screen platform, termed instant multi-color Fourier light-field microscopy (iFLFM), for simultaneous, all-optical assessment of heart morphology and function in order to identify causal genes of heart disease. The iFLFM system will be applied to monitor, in vivo, HLHS development in novel frog models of the disease. The Research Plan is developed under three aims: (1) Design, construct, and validate iFLFM to enable fast, volumetric, and simultaneous multi-color structural and dynamic imaging at the tissue and organ levels, which will form for basis for the instrumental platform for the proposed interrogation of embryonic frog hearts in vivo; (2) Validate iFLFM for in vivo observation of cardiac morphology and functions using control and Ets1-knockdown frog embryos, which will enable snapshot 3D and multi-parametric assessment and documentation of cardiac conditions and defects, confirm the influence of Ets1 as a causal gene for HLHS, and ready the basic architecture, assays, and processing for the proposed functional screen of other candidate genes; and (3) Advance the iFLFM platform for the functional screen of mutant embryonic frog hearts to identify causal genes for HLHS, which will reveal and document anatomical and functional variations and defects upon genetic mutations and identify mutations that lead to HLHS-like phenotype.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.