Non-invasive Aneuploidy Screening of Circulating Fetal Cells for Prenatal Diagnos

用于产前诊断的循环胎儿细胞的无创非整倍性筛查

基本信息

批准号：
8235596
负责人：
Matthew Rabinowitz
金额：
$ 80.07万
依托单位：
NATERA, INC.
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-08-15 至 2013-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8235596
关键词：
Adult Algorithms Alleles Amniocentesis Aneuploidy Antibodies Applications Grants Biopsy Blood Blood specimen Cell Count Cell Separation Cells Cheek structure Child Chorionic Villi Sampling Chromosome abnormality Chromosomes Clinical Clinical Research Clinical Trials Collection Comparative Study Confidence Intervals Congenital Abnormality Couples Coupling Custom Cytolysis DNA Data Density Gradient Centrifugation Detection Development Diagnosis Diagnostic Diagnostic tests Disabled Persons Disease Embryo Emotional Enrollment Erythroblasts Family Fathers Female Fertilization in Vitro Fetal Development Fetal Hemoglobin Fetal Tissues Fetus First Pregnancy Trimester General Population Genetic Genetic Screening Genome Genotype Gestational Age Goals Health Healthcare Hemoglobin Hereditary Disease Home environment Hour Individual Institutional Review Boards Karyotype Karyotype determination procedure Laboratories Lead Mainstreaming Measurement Measures Medical Medicine Metaphase Methods Metric Micromanipulation Modeling Molecular Mothers Nucleotides Outcome PTPRC gene Patients Performance Phase Physicians Pilot Projects Population Positioning Attribute Pregnancy Prenatal Diagnosis Procedures Process Protocols documentation Psyche structure Research Design Research Personnel Resolution Risk Sampling Screening procedure Second Pregnancy Trimester Sensitivity and Specificity Series Shipping Ships Single Nucleotide Polymorphism Site Solutions Sorting - Cell Movement Source Staining method Stains Statistical Computing Swab TFRC gene Techniques Technology Testing Time Tube Uniparental Disomy Validation Variant X Chromosome Y Chromosome base clinical practice density embryo stage 2 falls fetal fetal blood fetal diagnosis fetus cell follow-up genetic analysis genotyping technology handicapping condition improved innovation innovative technologies male pre-clinical preimplantation prenatal prevent prospective public health relevance research study sex single cell analysis success unborn child validation studies

项目摘要

DESCRIPTION (provided by applicant): During the course of a pregnancy, physicians and patients desire as much information as possible regarding the health of the fetus. For both emotional and medical reasons, this information is sought as early in term as possible, and with the fewest possible risks to both mother and child. Although the widely used first trimester chorionic villus sampling (CVS) and second trimester amniocentesis are relatively safe, both procedures are not without negligible risks. In efforts to avoid these risks altogether, researchers have turned toward isolating circulating fetal nucleated red blood cells (FNRBCs) from maternal blood as an alternative, non- invasive source of fetal tissue. Despite the development of FNRBC enrichment methods, there has been limited success with their coupling to subsequent aneuploidy screening and several challenges still must be overcome such as ability to test single fetal cells for 24-chromosome aneuploidy, confirm the isolated cell's origin (fetal versus maternal) and simultaneously screen for diseases caused by single nucleotide variants or micro in/dels. Our innovative Parental SupportTM technology provides a solution to all of these challenges and the development of a first trimester non-invasive prenatal diagnostic test is the ultimate goal of this grant application. In Phase I, we first plan to optimize single cell lysis and whole genome amplification protocols specifically for antibody-stained FNRBCs.. Protocol optimization for single cell analysis falls within the core competencies of GSN as we have previously successfully commercialized an innovative single cell molecular karyotyping protocol to enable genetic analysis of single blastomeres within 24 hours. We will then systematically evaluate which combination of existing FNRBC enrichment methods provides maximum yield and purity suitable for subsequent Parental Support"-based genetic analysis using predefined mixtures of fetal and adult blood. The main objective of Phase II will be to transition from the predefined blood mixtures of fetal and adult blood to actual maternal blood samples. We will first conduct a pilot study to determine which of the best FNRBC isolation method(s) identified in Phase I should become the lead method. Using this lead method, we will then conduct a larger study to evaluate concordance between aneuploidy diagnosis by Parental SupportTM and karyotyping by amniocentesis or chorionic villus sampling. If successful, we expect that the completion of these Aims would have a major impact on the field of prenatal diagnosis, improve the lives of millions of couples and children worldwide, and bring non-invasive diagnosis to the mainstream of prenatal medicine. PUBLIC HEALTH RELEVANCE: In the absence of prenatal diagnosis, up to 1 in 50 babies have serious physical or mental handicaps, up to 1 in 30 babies have some form of congenital malformation, and up to 1 in 200 have a phenotypically significant chromosome abnormality Although these abnormalities can be diagnosed with techniques such as amniocentesis or chorionic villus sampling, both procedures carry an increased risk of harm to both the mother and fetus. Our innovative technology has the potential to evaluate the health of an unborn child by simply analyzing the mother's blood, thereby minimizing the risks of the procedure and expanding prenatal screening to the general population.

描述（由申请人提供）：在怀孕期间，医生和患者希望有关胎儿健康的尽可能多的信息。由于情感和医学原因，这些信息在尽可能的早期都被寻求，并且对母子的风险最少。尽管广泛使用的妊娠中期绒毛膜绒毛采样（CVS）和妊娠中期羊膜穿刺术相对安全，但这两种程序并非没有可忽略的风险。为了避免这些风险，研究人员已转向从母体血液中隔离循环的胎儿成核红细胞（FNRBC），作为胎儿组织的替代性，非侵入性来源。尽管发展了FNRBC富集方法，但由于它们与随后的非整倍性筛查的耦合的成功有限，仍然必须克服几个挑战，例如能够测试单个胎儿细胞的24个染色体类肾上腺素的能力，确认孤立的细胞起源（胎儿对孕产妇）和单次分组的筛查序列或单个筛查的群体或单个核心的变化。我们创新的父母支持TM技术为所有这些挑战提供了解决方案，并且开发了孕期非侵入性产前诊断测试是该赠款应用的最终目标。在第一阶段，我们首先计划优化针对抗体染色的FNRBCS的单细胞裂解和整个基因组扩增方案。单细胞分析的方案优化属于GSN的核心竞争力，因为我们以前已经成功地将创新的单细胞分子型型协议商业化了，以启用单胚泡内24小时内24小时内的单个单细胞分子型方案。然后，我们将系统地评估现有的FNRBC富集方法的哪种组合可提供最大的产量和纯度，适合于随后的父母支持”基于胎儿和成人血液的预定混合物的基于遗传分析。第二阶段的主要目标是从胎儿和成人血液的预期血液混合物过渡到实际的血液样品，我们将最大程度地研究属于孕产妇。然后，我们将使用这种铅方法确定，我们将进行一项更大的研究，以评估父母支持的非整倍性诊断和羊膜中的核分型，如果成功的绒毛绒毛的完成，我们预计这些目标将对儿童的生命及其生命的影响，以改善PRENAT的生命。对产前医学主流的非侵入性诊断。 PUBLIC HEALTH RELEVANCE: In the absence of prenatal diagnosis, up to 1 in 50 babies have serious physical or mental handicaps, up to 1 in 30 babies have some form of congenital malformation, and up to 1 in 200 have a phenotypically significant chromosome abnormality Although these abnormalities can be diagnosed with techniques such as amniocentesis or chorionic villus sampling, both procedures carry an increased risk of harm to both the母亲和胎儿。我们的创新技术有可能通过简单地分析母亲的血液来评估未出生的孩子的健康，从而最大程度地减少程序的风险并扩大产前筛查向普通人群。