In utero mouse embryo phenotyping with high-frequency ultrasound

高频超声对小鼠子宫内胚胎表型分析

• 批准号: 9168204
• 负责人: Jeffrey Ketterling
• 金额: $ 75.78万
• 依托单位: RIVERSIDE RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-30 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9168204
• 关键词: Algorithms; Archives; Benchmarking; Brain; Cardiovascular system; Coitus; Computer software; Custom; Data; Data Analyses; Data Set; Databases; Defect; Detection; Development; Disease; Dissection; Embryo; Embryonic Development; Ensure; Evaluation; Eye; Frequencies; Funding; Future; Genital system; Goals; Growth; Head; Heart; Hour; Human; Image; International; Knock-out; Knockout Mice; Lateral; Limb structure; Liver; Longitudinal Studies; Lung; Methods; Modality; Modeling; Morphology; Mouse Strains; Mus; Optics; Perinatal; Phenotype; Process; Protocols documentation; Research; Resolution; Sampling; Staging; Staining method; Stains; Structural Congenital Anomalies; System; Testing; Three-Dimensional Image; Three-Dimensional Imaging; Time; Ultrasonography; United States National Institutes of Health; X-Ray Computed Tomography; abstracting; animal imaging; body system; data acquisition; fetal; image processing; imaging modality; in utero; in vivo; instrumentation; mutant; novel; programs; relating to nervous system; tomography; tool

Project Summary/Abstract The goal of this proposal is to phenotype early- to mid-gestational mouse embryos by segmenting select organ systems in 3D data sets acquired in utero with high-frequency ultrasound (HFU). The International Mouse Phe- notyping Consortium (IMPC), which includes the NIH Knockout (KO) Mouse Phenotyping Program (KOMP2), will generate 20,000 mouse strains in the next decade, including many important models of human structural birth defects and congenital diseases. The development of phenotyping methods that provide for efficient pipeline analyses of defects in embryonic growth in the KO mouse strains is a high priority for this effort. An in utero 3D imaging approach, enabling volumetric and longitudinal analyses of a variety of organ systems over a range of early- to mid-gestational stage mouse embryos, would provide added benefit and critical additional in vivo data not currently available. Commercial HFU systems are widely available in many research centers largely thanks to the NIH-funded Small Animal Imaging Research Programs and Shared Instrumentation Programs. HFU is therefore an excellent candidate modality to provide in utero 3D image data that can be quantitatively analyzed and archived to support the KOMP2/IMPC embryonic lethal phenotyping pipeline and future phenotyping efforts. We propose to develop and validate in utero 3D HFU image-acquisition protocols and image-processing meth- ods that permit noninvasive, longitudinal studies of embryonic development and, in particular, the detection and characterization of KO phenotypes. Volumetric HFU data will be collected in utero from mouse embryos staged between E9.5 to 15.5 in order to establish a database of normal development. Algorithms will be developed to segment 3D regions and extract parameters that quantify embryonic stage and identify regional changes between normal and KO embryos. We will acquire data with a custom, annular-array system and with a VisualSonics Vevo 2100. The fine-resolution annular-array data will be used to initially develop the image-processing algorithms and then the algorithms will be adapted for Vevo 2100 data. We will compare the quantitative parameters derived from the segmentation results obtained from the two scanners to ensure that the Vevo 2100 is able to provide equivalent mutant detection and quantification. Initial testing will be undertaken using wild-type and En1 and Gli2 mutants that have known defects. Finally, the acquisition and processing protocols will be applied to 3D Vevo 2100 data from 5-10 KOMP2 KO mouse lines with embryonic defects in a variety of organ systems to validate the HFU methods for detecting and characterizing phenotypes in these mutant embryos.

项目概要/摘要 该提案的目标是通过分割选定的器官来对妊娠早期至中期小鼠胚胎进行表型分析 使用高频超声 (HFU) 在子宫内采集的 3D 数据集系统。 notyping 联盟 (IMPC)，其中包括 NIH 基因敲除 (KO) 小鼠表型分析计划 (KOMP2)，将 在未来十年内产生 20,000 个小鼠品系，其中包括许多重要的人类结构出生模型 缺陷和先天性疾病的表型分析方法的开发提供了有效的管道。 分析 KO 小鼠品系的胚胎生长缺陷是这项工作的首要任务。 成像方法，能够对一系列器官系统进行体积和纵向分析 妊娠早期到中期的小鼠胚胎，将提供额外的益处和关键的额外体内数据 目前尚未在许多研究中心广泛使用商业 HFU 系统，这主要归功于 NIH 资助的小动物成像研究计划和共享仪器计划。 因此，这是一种提供可定量分析的子宫内 3D 图像数据的绝佳候选方式 并存档以支持 KOMP2/IMPC 胚胎致死表型分析流程和未来的表型分析工作。 我们建议开发并验证子宫内 3D HFU 图像采集协议和图像处理方法 ODS 允许对胚胎发育进行非侵入性纵向研究，特别是检测和 KO 表型的表征将从子宫内分期的小鼠胚胎中收集体积 HFU 数据。 E9.5到15.5之间为了建立正常开发的算法将被开发。 分割 3D 区域并提取量化胚胎阶段并识别胚胎阶段之间区域变化的参数 我们将使用定制的环形阵列系统和 VisualSonics Vevo 采集数据。 2100. 高分辨率环形阵列数据将用于初步开发图像处理算法和 然后算法将适用于 Vevo 2100 数据，我们将比较导出的定量参数。 根据两台扫描仪获得的分割结果，确保 Vevo 2100 能够提供 等效突变体检测和定量将使用野生型以及 En1 和 Gli2 进行。 最后，采集和处理协议将应用于 3D Vevo。 来自 5-10 个 KOMP2 KO 小鼠品系的 2100 个数据，这些小鼠品系在多种器官系统中具有胚胎缺陷，以验证 用于检测和表征这些突变胚胎表型的 HFU 方法。