Journal of Assisted Reproduction and Genetics

, Volume 15, Issue 3, pp 140–144 | Cite as

Preimplantation Genetic Diagnosis of Inherited Cancer: Familial Adenomatous Polyposis Coli

  • Asangla Ao
  • Dagan Wells
  • Alan H. Handyside
  • Robert M. L. Winston
  • Joy D. A. Delhanty


Purpose:Our purpose was to achieve preimplantation genetic diagnosis (PGD) of the dominant cancer predisposition syndrome, familial adenomatous polyposis coli (FAPC), as an alternative to prenatal diagnosis.

Methods:The affected patient was superovulated and oocytes were retrieved and fertilized by intracytoplasmic sperm injection (ICSI). Two cells were biopsiedfrom each embryo and the whole genome was amplified by primer extension preamplification (PEP). Nested PCR was then used to amplify two APC fragments: one including the APC mutation site and the other an informative intragenic polymorphism. Both were detected by simultaneous singlestrand conformation polymorphism and heteroduplex analysis.

Results:Four normally fertilized embryos were biopsied on day 3 post ICSI, and two cells were successfully removed from each embryo. Following PEP the APC mutation was successfully amplified in 7 of 8 cells, and the polymorphism in 6 of 8 cells. The APC mutation was detected in three embryos. This result was confirmed by identification of the mutation associated polymorphism in two cases. A single embryo was diagnosed as homozygous normal for the mutation and the polymorphism in both cells sampled. This unaffected embryo was transferred to the mother, but no pregnancy resulted.

Conclusions:We report here the first diagnosis of a cancer predisposition syndrome in human preimplantation embryos. Our results indicate that difficulties associated with single-cell PCR, allele-specific amplification failure in particular, need not prevent preimplantation diagnosis of diseases with a dominant mode of inheritance, provided appropriate strategies are applied.

familial adenomatous polyposis coli cancer predisposition preimplantation diagnosis 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Bryant H: Genetic screening for breast cancer in Ashkenazi women. Lancet 1996;347:1638–1639Google Scholar
  2. 2.
    Ao A, Handyside AH: Cleavage stage human embryo biopsy. Hum Reprod Update 1995;1:3Google Scholar
  3. 3.
    Handyside AH, Kontogianni EH, Hardy K, Winston RM: Pregnancies from biopsied human preimplantation embryos sexed by Y-specific DNA amplification. Nature 1990;344:768–770Google Scholar
  4. 4.
    Harper JC, Handyside AH: The current status of preimplantation diagnosis. Curr Obstet Gynecol 1994;4:143–149Google Scholar
  5. 5.
    Harton GL, Tsipouras P, Sisson ME, et al.: Preimplantation genetic testing for Marfan syndrome. Mol Hum Reprod 1996;2:713–715Google Scholar
  6. 6.
    Herrera L, Kakaki S, Gibas L, Pietrzak K, Sandberg AA: Brief clinical report: Gardner syndrome in a man with an intestitial deletion of 5q. Am J Med Genet 1986;25:473–476Google Scholar
  7. 7.
    Groden J, Thliveris A, Samowitz W, et al.: Identification and characterization of the familial adenomatous polyposis coli gene. Cell 1996;66(3):589–600Google Scholar
  8. 8.
    Kinzler KW, Nilbert MC, Su LK, et al.: Identification of FAP locus genes from chromosome 5q21. Science 1991;253:661–665Google Scholar
  9. 9.
    Whitelaw S, Northover JM, Hodgson SV: Attitudes to predictive DNA testing in familial adenomatous polyposis. J Med Genet 1996;33:540–543Google Scholar
  10. 10.
    Gayther SA, Wells D, Sen Gupta SB, et al.: Regionally clustered APC mutations are associated with a severe phenotype and occur at a high frequency in new mutation cases of adenomatous polyposis coli. Hum Mol Genet 1994;3:53–56Google Scholar
  11. 11.
    Gayther SA, Wells D, Gulati K, Chapman P, Burn J, Delhanty JDA: Germline rearrangement of MCC and APC detected by pulsed field gel electrophoresis and fluorescent in situ hybridization. Ann Hum Genet 1993;57:169–178Google Scholar
  12. 12.
    Findlay I, Ray P, Quirke P, Rutherford A, Lilford R: Allelic drop-out and preferential amplification in single cells and human blastomeres: Implications for preimplantation diagnosis of sex and cystic fibrosis. Hum Reprod 1995;10:1609–1618Google Scholar
  13. 13.
    Ray PF, Handyside AH: Increasing the denaturation temperature during the first cycles of amplification reduces allele dropout from single cells for preimplantation genetic diagnosis. Mol Hum Reprod 1996;2(3):213–218Google Scholar
  14. 14.
    Nagase H, Miyoshi Y, Horri A, et al.: Screening for germline mutations in familial adenomatous polyposis patients: 61 new patients and a summary of 150 unrelated patients. Hum Mutat 1992;1:467–473Google Scholar
  15. 15.
    Zhang L, Cui X, Schmitt K, Hubert R, Navidi W, Arnheim N: Whole genome amplification from a single cell: Implications for genetic analysis. Proc Natl Acad Sci USA 1992;89: 5847–5851Google Scholar
  16. 16.
    Rutherford AJ, Subak Sharpe RJ, Dawson KJ, Margara RA, Franks S, Winston RM: Improvement of in vitro fertilisation after treatment with buserelin, an agonist of luteinising hormone releasing hormone. B M J 1988;296: 1765–1768Google Scholar
  17. 17.
    Cui XF, Li HH, Goradia TM, et al.: Single-sperm typing: Determination of genetic distance between the G gamma-globin and parathyroid hormone loci by using the polymerase chain reaction and allele-specific oligomers. Proc Natl Acad Sci USA 1989;86:9389–9393Google Scholar
  18. 18.
    Handyside AH, John LG, Tarin JJ, Winston RML, Hughes MR: Birth of a normal girl after in vitro fertilization and preimplantation diagnostic testing for cystic fibrosis. N Engl J Med 1992;327:905–909Google Scholar
  19. 19.
    Giorgetti C, Terriou P, Auquier P, et al.: Embryo score to predict implantation after in vitro fertilization: Based on 957 single embryo transfer. Hum Reprod 1995;10:2427–2431Google Scholar
  20. 20.
    Ao A, Ray P, Harper JC, et al.: Clinical experience with preimplantation diagnosis of cystic fibrosis (ΔF508). Prenat Digan 1996;16:137–142Google Scholar
  21. 21.
    Struewing JP, Watson P, Easton DF, Ponder BA, Lynch HT, Tucker MA: Prophylactic oophorectomy in inherited breast/ovarian cancer families. J Natl Cancer Inst Monogr 1995;17:33–35Google Scholar

Copyright information

© Plenum Publishing Corporation 1998

Authors and Affiliations

  • Asangla Ao
    • 1
  • Dagan Wells
    • 2
  • Alan H. Handyside
    • 3
  • Robert M. L. Winston
    • 1
  • Joy D. A. Delhanty
    • 2
  1. 1.Institute of Obstetrics and Gynaecology, RPMSHammersmith HospitalLondonUK
  2. 2.Human Genetics Group, Gallon LaboratoryUniversity College LondonLondonUK
  3. 3.Department of Obstetrics & GynaecologySt.Thomas' HospitalLondonUK

Personalised recommendations