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：
7910271
负责人：
Matthew Rabinowitz
金额：
$ 20.08万
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 2011-02-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7910271
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 Karyotype Karyotype determination procedure Laboratories Lead Life 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 Ethics Committees 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 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，医生和患者希望获得尽可能多的有关胎儿健康的信息。出于情感和医学原因，应在足月时尽早寻求这些信息，以尽量减少对母亲和孩子的风险。尽管广泛使用的孕早期绒毛膜绒毛取样（CVS）和孕中期羊膜穿刺术相对安全，但这两种手术并非没有可忽略的风险。为了完全避免这些风险，研究人员转向从母血中分离循环胎儿有核红细胞 (FNRBC)，作为胎儿组织的替代非侵入性来源。尽管 FNRBC 富集方法得到了发展，但其与后续非整倍体筛选的结合取得的成功有限，并且仍然必须克服一些挑战，例如测试单个胎儿细胞的 24 染色体非整倍性的能力、确认分离细胞的起源（胎儿与母体细胞））并同时筛查由单核苷酸变异或微插入/缺失引起的疾病。我们创新的 Parental SupportTM 技术为所有这些挑战提供了解决方案，开发妊娠早期无创产前诊断测试是本次拨款申请的最终目标。在第一阶段，我们首先计划专门针对抗体染色的 FNRBC 优化单细胞裂解和全基因组扩增方案。单细胞分析方案优化属于 GSN 的核心能力，因为我们之前已成功将创新的单细胞分子核型分析商业化协议能够在 24 小时内对单个卵裂球进行遗传分析。然后，我们将系统地评估现有 FNRBC 富集方法的哪种组合可提供最大产量和纯度，适合随后使用胎儿和成人血液的预定义混合物进行基于“父母支持”的遗传分析。第二阶段的主要目标是从预定义的血液过渡我们将首先进行一项试点研究，以确定第一阶段确定的最佳 FNRBC 分离方法应成为主要方法，然后我们将使用这种主要方法进行。进行更大规模的研究来评估Parental SupportTM 的非整倍体诊断与羊膜穿刺术或绒毛膜绒毛取样的核型分析之间的一致性如果成功，我们预计这些目标的完成将对产前诊断领域产生重大影响，改善全世界数百万夫妇和儿童的生活，并将无创诊断带入产前医学主流。公共卫生相关性：在没有产前诊断的情况下，多达五分之一的婴儿患有严重的身体或精神障碍，多达三十分之一的婴儿患有某种形式的先天畸形，多达二分之一的婴儿患有表型显着的染色体异常。这些异常可以通过羊膜穿刺术或绒毛膜绒毛取样等技术来诊断，这两种手术都会增加对母亲和母亲造成伤害的风险。胎儿。我们的创新技术有潜力通过简单地分析母亲的血液来评估未出生婴儿的健康状况，从而最大限度地降低手术风险并将产前筛查扩大到普通人群。