Preimplantation Genetic Diagnosis (PGD) for Fanconi Anemia and HLA Christina M. Lavin, MS, CGC Licensed Genetic Counselor Reproductive Genetics Institute (RGI) Chicago, IL USA June 28, 2010 OBJECTIVES Preimplantation Genetic Diagnosis (PGD)  In Vitro Fertilization (IVF)  Fees / Insurance  Common Questions  Success / RGI experience with FA  Advantages / Disadvantages / Limitations  Definitions: Preimplantation Genetic Diagnosis (PGD): Diagnosis of Genetic Disease Before Pregnancy In Vitro Fertilization (IVF): Assisted Reproductive technology procedure where oocytes (eggs) are removed from a woman’s ovary and are fertilized by the male’s sperm outside the body in a controlled setting. The fertilized eggs develop into embryos which can be implanted in a woman’s uterus to achieve pregnancy. Current indications for PGD: *Autosomal Recessive, Dominant and X liked Disorders * HLA matching * Cancer predisposition genes * Adult-onset disorders * Infertility-causing genes * Maternal-fetal incompatibility * Chromosomal Aneuploidy * Combined Single gene, HLA, and aneuploidy testing Testing Techniques utilized in PGD: PCR (polymerase chain reaction)  Single gene (Mendelian) disorders like FA  Mutation(s) with linked markers and HLA matching  FISH (fluorescent in-situ hybridization)  Aneuploidy (abnormal # of chromosomes)  Translocations/structural rearrangements  Getting started…. PGD Set-up  Create a testing system for the  specific FA mutations and HLA Prepare for IVF  PGD SET-UP for FA/HLA MEDICAL RECORDS: FA mutation reports, HLA reports  PGD CONSULT (phone or in person) with a PGD  genetic counselor. Discuss the benefits, risks, limitations, accuracy (95-98% for FA and HLA) etc….. DNA SAMPLES (blood or cheek swabs) from parents  and children CONSENT FORMS  FEES  SET-UP SYSTEM takes 4-6 weeks to complete  Testing for mutations and linked markers  TESTING STRATEGY (PB/BB vs. BB)  IVF CYCLE DATES!!!!!!  Reproductive Genetics Institute FANCONI ANEMIA-A LINKED MARKERS 1. D16S520 PGD SET-UP 2. D16S3026 3. FANCA Intron 1 4. FANCA (Paternal Mutation :M) 5. FANCA (Maternal Mutation: M) 6. D16S3407 Husband Wife (carrier) (carrier) 1. 10 / a 1. g / 70 2. 20 / b 2. h/ 80 3. 30 / c 3. i / 90 4. M / N 4. N / N Child with FA 5. N / N 5. N / M 6. 60 / f 6. K / 120 1. 10 / 70 2. 20 / 80 3. 30 / 90 4. M / N 5. N / M 6. 60 / 120 Telomere D6S461 Updated (June 2007) List of D6S276* D6S299 Markers in HLA Area for D6S464* D6S105 Preimplantation HLA Typing HLA-F D6S306* D6S1624* HLA-A D6S1615 D6S258 D6S248* HLA-E I s MOG a,b,c,d s RF a HLA-C lC D6S265 D6S510 HLA-B MIB MICA TNF a,b,c,d III s 6822-1 s a 9 N2 6p21.3 lC D6S273* D6S1666 D6S1629 DN HLA-DR LH 1 DQ-CAR II HLA-DQ DQ-CAR G51152 II s D6S2447 s a D6S2443 lC D6S2444 TAP 1 Ring 3CA D6S1568 D6S1560 D6S1618 D6S439 D6S291 D6S1583 D6S1610 Centromere Why are Linked Markers so important? * Detection of Allele Drop Out (ADO) * Detection of Recombination * Detection of contamination * Detection of chromosomal aneuploidy * Embryo identification Reproductive Genetics Institute