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.
Thesaurus Terms: 0-11 Years Old;21+ Years Old;Aberrant Chromosome;Abnormalities, Chromosomal;Active Follow-Up;Adult;Algorithms;Alleles;Allelomorphs;Amniocentesis;Aneuploid;Aneuploidy;Antibodies;Applications Grants;B220;Biopsy;Biopsy, Chorionic Villi;Birth Defects;Blastocytes;Blastomere;Blood;Blood Sample;Blood Specimen;Bucca;Buccas;Cd45;Cd71;Cvs;Care, Health;Cell Count;Cell Isolation;Cell Number;Cell Segregation;Cell Separation;Cell Separation Technology;Cells;Centrifugation, Density Gradient;Cheek;Cheek Structure;Child;Child Youth;Children (0-21);Chorionic Villi Sampling;Chromosomal Aberrations;Chromosomal Alterations;Chromosome Aberrations;Chromosome Alterations;Chromosome Anomalies;Chromosome Abnormality;Chromosomes;Clinical;Clinical Research;Clinical Study;Clinical Trials;Clinical Trials, Unspecified;Collection;Comparative Study;Computing, Statistical;Confidence Intervals;Congenital Abnormality;Congenital Anatomic Abnormality;Congenital Anatomical Abnormality;Congenital Defects;Congenital Deformity;Congenital Malformation;Couples;Coupling;Custom;Cytogenetic Aberrations;Cytogenetic Abnormalities;Cytolysis;Dna;Data;Density Gradient Centrifugation;Deoxyribonucleic Acid;Detection;Development;Diagnosis;Diagnosis, Antenatal;Diagnostic;Diagnostic Tests;Disabled Persons;Disabled Population;Disease;Disorder;Early Placental Phase;Educational Mainstreaming;Embryo;Embryo Stage 2;Embryonic;Emotional;Enrollment;Erythroblasts;Erythrocytes, Nucleated;Ethics Committees, Research;Family;Fathers;Female;Fertilization In Vitro;Fetal Age;Fetal Development;Fetal Hemoglobin;Fetal Maturity, Chronologic;Fetal Tissues;Fetal Development Of The Mammalian Embryo Or Fetus;Fetus;First Pregnancy Trimester;Fractionation, Density Gradient;Gp180;General Population;General Public;Genetic;Genetic Condition;Genetic Diseases;Genetic Screening;Genetic Analyses;Genetics, Karyotyping;Genome;Genotype;Gestation;Gestational Age;Goals;Grant Proposals;Grants, Applications;Handicapped;Health;Healthcare;Hemoglobin;Hemoglobin F;Hereditary Disease;Home;Home Environment;Hour;Human, Adult;Human, Child;Irbs;Ivf;Individual;Infirmity;Institutional Review Boards;Intrauterine Diagnosis;Investigators;Karyotype;Karyotype Determination Procedure;Karyotyping;Lca;Ly5;Laboratories;Lead;Life;Lysis;Mainstream Education, Achievement;Mainstreaming;Mainstreaming (Education);Measurement;Measures;Medical;Medicine;Metaphase;Methods;Methods And Techniques;Methods, Other;Metric;Micromanipulation;Midtrimester;Mitotic Metaphase;Modeling;Molecular;Molecular Disease;Molecular Genetic Abnormality;Mothers;Normoblasts;Nucleated Red Blood Cell;Nucleated Red Cell;Nucleotides;Outcome;Ptprc;Ptprc Gene;Patients;Pb Element;People With Disabilities;Performance;Persons With Disabilities;Phase;Physicians;Pilot Projects;Polymorphism, Single Base;Population;Position;Positioning Attribute;Pregnancy;Pregnancy Trimester, First;Pregnancy Trimester, Second;Prenatal Diagnosis;Procedures;Process;Protocol;Protocols Documentation;Psyche Structure;Research Design;Research Ethics Committees;Research Personnel;Researchers;Resolution;Reticuloendothelial System, Blood;Risk;Snp;Snps;Sampling;Science Of Medicine;Screening Procedure;Second Pregnancy Trimester;Sensitivity And Specificity;Series;Shipping;Ships;Single Nucleotide Polymorphism;Site;Solutions;Sorting - Cell Movement;Source;Staining Method;Stainings;Stains;Statistical Computing;Statistical Computings;Structure Of Embryo Stage 2;Study Type;Swab;T200;Tfr;Tfrc;Tfrc Gene;Trfr;Techniques;Technology;Test-Tube Fertilization;Testing;Time;Tissues, Fetal;Trimester, First;Trimester, Second;Tube;Uniparental Disomy;Validation;Variant;Variation;Y Chromosome;Adult Human (21+);Antepartum Diagnosis;Base;Cell Sorting;Children;Clinical Investigation;Clinical Practice;Density;Density Gradient Ultracentrifugation;Disabled;Disabled People;Disease/Disorder;Embryo Stage 2;Enroll;Experiment;Experimental Research;Experimental Study;Falls;Fetal;Fetal Blood;Fetal Diagnosis;Fetus Cell;Fetus Tissue;Follow-Up;Genetic Analysis;Genetic Disorder;Genotyping Technology;Handicapping;Handicapping Condition;Heavy Metal Pb;Heavy Metal Lead;Hereditary Disorder;Improved;Innovate;Innovation;Innovative;Innovative Technologies;Karyogram;Male;Mental;Nucleated Rbcs;Pilot Study;Pre-Clinical;Preclinical;Preimplantation;Prenatal;Prevent;Preventing;Prospective;Public Health Relevance;Research Study;Screening;Screenings;Sex;Single Cell Analysis;Sorting;Study Design;Success;Unborn;Unborn Child;Validation Studies;Youngster
