Non-Invasive Prenatal Diagnostics Based on Circulating Trophoblasts

基于循环滋养细胞的无创产前诊断

基本信息

批准号：
10675005
负责人：
Margareta Pisarska
金额：
$ 79.3万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-15 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10675005
关键词：
Acceleration Adopted Advanced Development Amniocentesis Aneuploidy Biological Markers Biological Models Biometry Blood Blood Cells Blood specimen Cell Separation Cells Cervix Uteri Chemistry Chorionic Villi Sampling Chromosome abnormality Clinic Clinical Collaborations Collecting Cell Collection Computer software DNA DNA amplification Data Detection Diagnosis Diagnostic Diagnostic tests Discipline of obstetrics Down Syndrome Exclusion Exhibits Fetus First Pregnancy Trimester Funding Genetic Diseases Genome Genomic DNA Genotype Gestational Age Goals Harvest In Vitro Individual Interdisciplinary Study Joints Karyotype Karyotype determination procedure Longevity Medical center Methods Microarray Analysis Microfluidics Molecular Abnormality Monitor Motivation NanoVelcro Nanostructures Neoplasm Circulating Cells Pathology Performance Pregnancy Pregnant Women Prenatal Diagnosis Procedures Process Public Health Reproducibility Research Resolution Risk Sampling Sensitivity and Specificity Spontaneous abortion Surface Swab Testing Validation biomarker signature cell free fetal DNA clinical application cohort cost efficient diagnostic platform diagnostic strategy diagnostic technologies diagnostic value fabrication feasibility testing fetal fetal diagnosis fetus cell genetic disorder diagnosis genetic testing genome-wide immunocytochemistry imprint improved innovation laser capture microdissection microdeletion microscopic imaging nanoimprinting nanomaterials next generation sequencing prenatal prenatal testing preservation recruit screening single cell analysis specific biomarkers success trophoblast whole genome

项目摘要

PROJECT SUMMARY/ABSTRACT The long-term goal of this revised U01 proposal is to conduct advanced development and rigorous validation of an emerging circulating trophoblast (cTB)-based noninvasive prenatal diagnostic (NIPD) technology, capable of i) enriching/counting cTBs from maternal blood, and ii) isolating single cTBs for genome-wide detection of fetal genetic abnormalities during the first trimester of pregnancy. An alternative research plan is also presented to explore the use of the same workflow for isolating and characterizing trophoblasts (TBs) in cervix samples. Among potential circulating fetal nucleated cells (CFNCs) in maternal blood, cTBs are an ideal target considering their (i) short lifespan, which excludes the presence of cTBs from prior pregnancies or miscarriages, (ii) representation of fetal karyotype and genotype, and (iii) expression of a unique collection of biomarkers that can be used for both enrichment and identification. However, isolating pure cTBs has been technically challenging due to their extremely low abundance. Over the past decade, Dr. Tseng’s research team at UCLA has developed nanomaterial-embedded diagnostic platforms (a.k.a., NanoVelcro Chips). To exploit the NIPD utility of NanoVelcro Chips, the team first developed a nanoimprinting fabrication process to prepare the laser capture microdissection (LCM)-compatible nanosubstrates in a cost-efficient and scalable manner. These chips, in conjunction with the use of capture and immunocytochemistry (ICC) agents, exhibit superb cTB capture performance. In parallel, high-resolution microscopy imaging and analysis software has been developed to identify and register individual cTBs on the substrates, enabling highly accurate isolation of single cTBs by LCM. In collaboration with Dr. Pisarska, the joint team demonstrated a workflow starting with blood processing, single cTB isolation, and DNA amplification, all the way through whole genome profiling of cTBs by ArrayCGH and/or next generation sequencing. Our central hypothesis is that >10 cTBs can be harvested from 5-mL of maternal blood (>50 TBs from a cervix sample), collected from a pregnant woman during the first trimester of pregnancy (8-12 weeks of gestational age), and whole genome profiling of these cTBs/TBs can be used for diagnosing fetal genetic abnormalities. Over the 5-year funding period, the proposed research will be implemented via two Specific Aims: i) to develop, optimize and validate the proposed cTB-based NIPD technology, and ii) to conduct initial clinical validation in pregnant women recruited from UCLA and CSMC. The joint team envisions that the successful demonstration of the proposed cTBs-based NIPD technology will introduce a revolutionary NIPD solution with the sensitivity and specificity of the gold standard diagnostic tests without the associated risks to the fetus.

项目概要/摘要此次修订的 U01 提案的长期目标是对一种新兴的基于循环滋养层（cTB）的无创产前诊断（NIPD）技术，能够 i) 从母血中富集/计数 cTB，以及 ii) 分离单个 cTB 以进行全基因组检测怀孕前三个月的胎儿遗传异常也是一个替代研究计划。旨在探索使用相同的工作流程来分离和表征子宫颈中的滋养层细胞 (TB) 样品。在母血中潜在的循环胎儿有核细胞 (CFNC) 中，cTB 是理想的靶标 (i) 寿命短，不包括先前怀孕中存在的 cTB 或流产，(ii) 胎儿核型和基因型的表示，以及 (iii) 独特集合的表达可用于富集和鉴定的生物标志物然而，分离纯 cTB 一直是个难题。由于丰度极低，技术上具有挑战性。在过去的十年里，曾博士在加州大学洛杉矶分校的研究团队开发了纳米材料嵌入诊断平台（又名 NanoVelcro 芯片）为了利用 NanoVelcro 芯片的 NIPD 实用程序，该团队首先开发了一种纳米压印制造工艺来制备兼容激光捕获显微切割（LCM）的这些芯片以经济高效且可扩展的方式结合捕获和使用。免疫细胞化学 (ICC) 试剂具有出色的并行、高分辨率 cTB 捕获性能。显微成像和分析软件已开发用于识别和注册单个 cTB 与 Pisarska 博士合作，能够通过 LCM 高精度分离单个 cTB。联合团队展示了从血液处理、单一 cTB 分离和 DNA 扩增开始的工作流程，通过 ArrayCGH 和/或下一代测序对 cTB 进行全基因组分析。我们的中心假设是，可以从 5 mL 母血中收获 > 10 个 cTB（从母体血液中收获 > 50 个 TB）。子宫颈样本），从怀孕前三个月（怀孕 8-12 周）的孕妇采集孕龄），这些 cTB/TB 的全基因组分析可用于诊断胎儿遗传在5年的资助期内，拟议的研究将通过两个具体项目实施。目标：i) 开发、优化和验证所提议的基于 cTB 的 NIPD 技术，以及 ii) 进行初步研究在从 UCLA 和 CSMC 招募的孕妇中进行临床验证联合团队预计，所提议的基于 cTB 的 NIPD 技术的成功演示将带来革命性的 NIPD 解决方案具有金标准诊断测试的敏感性和特异性，且不存在相关风险胎儿。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

High-throughput mRNA-seq atlas of human placenta shows vast transcriptome remodeling from first to third trimester.

人类胎盘的高通量 mRNA-seq 图谱显示从妊娠早期到晚期的巨大转录组重塑。

DOI：
10.1101/2023.06.06.543972
发表时间：
2023
期刊：
bioRxiv : the preprint server for biology
影响因子：
0
作者：
Gonzalez,TaniaL;Wertheimer,Sahar;Flowers,AmyE;Wang,Yizhou;Santiskulvong,Chintda;Clark,EkaterinaL;Jefferies,CarolineA;Lawrenson,Kate;Chan,JessicaL;Joshi,NikhilV;Zhu,Yazhen;Tseng,Hsian-Rong;Karumanchi,SAnanth;Williams,John;
通讯作者：