Monitoring for Potential Adverse Effects of Prenatal Gene Therapy: Use of Large Animal Models with Relevance to Human Application

  • Vedanta Mehta
  • Khalil N. Abi-Nader
  • David Carr
  • Jacqueline Wallace
  • Charles Coutelle
  • Simon N. Waddington
  • Donald PeeblesEmail author
  • Anna L. David
Part of the Methods in Molecular Biology book series (MIMB, volume 891)


Safety is an absolute prerequisite for introducing any new therapy, and the need to monitor the consequences of administration of both vector and transgene to the fetus is particularly important. The unique features of fetal development that make it an attractive target for gene therapy, such as its immature immune system and rapidly dividing populations of stem cells, also mean that small perturbations in pregnancy can have significant short- and long-term consequences. Certain features of the viral vectors used, the product of the delivered gene, and sometimes the invasive techniques necessary to deliver the construct to the fetus in utero have the potential to do harm.

An important goal of prenatal gene therapy research is to develop clinically relevant techniques that could be applied to cure or ameliorate human disease in utero on large animal models such as sheep or nonhuman primates. Equally important is the use of these models to monitor for potential adverse effects of such interventions. These large animal models provide good representation of individual patient-based investigations. However, analyses that require defined genetic backgrounds, high throughput, defined variability and statistical analyses, e.g. for initial studies on teratogenic and oncogenic effects, are best performed on larger groups of small animals, in particular mice.

This chapter gives an overview of the potential adverse effects in relation to prenatal gene therapy and describes the techniques that can be used experimentally in a large animal model to monitor the potential adverse consequences of prenatal gene therapy, with relevance to clinical application. The sheep model is particularly useful to allow serial monitoring of fetal growth and well-being after delivery of prenatal gene therapy. It is also amenable to serially sampling using minimally invasive and clinically relevant techniques such as ultrasound-guided blood sampling. For more invasive long-term monitoring, we describe telemetric techniques to measure the haemodynamics of the mother or fetus, for example, that interferes minimally with normal animal behaviour. Implanted catheters can also be used for serial fetal blood sampling during gestation. Finally, we describe methods to monitor events around birth and long-term neonatal follow-up that are important when considering human translation of this therapy.

Key words

Ultrasound Doppler Fetal growth velocity Amniocentesis Fetal blood sampling Blood flow Blood pressure Heart rate Neonate Miscarriage Liver biopsy Bone marrow biopsy 


  1. 1.
    Tabor A, Philip J, Madsen M et al (1986) Randomised controlled trial of genetic amniocentesis in 4606 low-risk women. Lancet 1:1287–1293PubMedCrossRefGoogle Scholar
  2. 2.
    Alfirevic Z, Sundberg K, Brigham S (2003) Amniocentesis and chorionic villus sampling for prenatal diagnosis. Cochrane Database of Systematic Reviews CD003252Google Scholar
  3. 3.
    Weiner CP, Wenstrom KD, Sipes SL, Williamson RA (1991) Risk factors for cordocentesis and fetal intravascular transfusion. Am J Obstet Gynecol 165:1020–1025PubMedGoogle Scholar
  4. 4.
    Peckham CS, Martin JA, Marshall WC, Dudgeon JA (1979) Congenital rubella deafness: a preventable disease. Lancet 8110: 258–261CrossRefGoogle Scholar
  5. 5.
    Preece PM, Pearl KN, Peckham CS (1984) Congenital cytomegalovirus infection. Arch Dis Child 59:1120–1126PubMedCrossRefGoogle Scholar
  6. 6.
    Wenstrom KD, Andrews WW, Bowles NE et al (1998) Intrauterine viral infection at the time of second trimester genetic amniocentesis. Obstet Gynaecol 92:420–424CrossRefGoogle Scholar
  7. 7.
    Dodic M, May CN, Wintour EM, Coghlan JP (1998) An early prenatal exposure to excess glucocorticoid leads to hypertensive offspring in sheep. Clin Sci 94:149–155PubMedGoogle Scholar
  8. 8.
    Schellenberg JC, Liggins GC (1987) New approaches to hormonal acceleration of fetal lung maturation. J Perinat Med 15:447–452PubMedCrossRefGoogle Scholar
  9. 9.
    Barker DJ, Osmond C, Simmonds SJ, Wield GA (1993) The relation of small head circumference and thinness at birth to death from cardiovascular disease in adult life. BMJ 306:422–426PubMedCrossRefGoogle Scholar
  10. 10.
    Miller SL, Loose JM, Jenkin G, Wallace EM (2008) The effects of sildenafil citrate (Viagra) on uterine blood flow and well being in the intrauterine growth-restricted fetus. Am J Obstet Gynaecol 102:e1–e7Google Scholar
  11. 11.
    David AL, Torondel B, Zachary I et al (2008) Local delivery of VEGF adenovirus to the uterine artery increases vasorelaxation and uterine blood flow in the pregnant sheep. Gene Ther 15:1344–1350PubMedCrossRefGoogle Scholar
  12. 12.
    Porada CD, Park PJ, Tellez J et al (2005) Male germ-line cells are at risk following direct-injection retroviral-mediated gene transfer in utero. Mol Ther 12:754–762PubMedCrossRefGoogle Scholar
  13. 13.
    Heikkilä A, Hiltunen MO, Turunen MP et al (2001) Angiographically guided utero-placental gene transfer in rabbits with adenoviruses, plasmid/liposomes and plasmid/polyethyleneimine complexes. Gene Ther 8:784–788PubMedCrossRefGoogle Scholar
  14. 14.
    MacCalman CD, Furth EE, Omigbodun A et al (1996) Transduction of human trophoblast cells by recombinant adenoviruses is differentiation dependent. Biol Reprod 54: 682–691PubMedCrossRefGoogle Scholar
  15. 15.
    Parry S, Holder J, Strauss JR (1997) Mechanisms of trophoblast-virus interaction. J Reprod Immunol 37:25–34PubMedCrossRefGoogle Scholar
  16. 16.
    Koi H, Zhang J, Makrigiannakis A et al (2001) Differential expression of the coxsackievirus and adenovirus receptor regulates adenovirus infection of the placenta. Biol Reprod 64:1001–1009PubMedCrossRefGoogle Scholar
  17. 17.
    Bedrosian JC, Gratton MA, Brigande JV et al (2006) In vivo delivery of recombinant viruses to the fetal murine cochlea: transduction characteristics and long-term effects on auditory function. Mol Ther 14:328–335PubMedCrossRefGoogle Scholar
  18. 18.
    Brown AS, Derkits EJ (2010) Prenatal infection and schizophrenia: a review of epidemiologic and translational studies. Am J Psychiatry 167:261–280PubMedCrossRefGoogle Scholar
  19. 19.
    Themis M, Waddington SN, Schmidt M et al (2005) Oncogenesis following delivery of a nonprimate lentiviral gene therapy vector to fetal and neonatal mice. Mol Ther 12: 763–771PubMedCrossRefGoogle Scholar
  20. 20.
    Wong LF, Goodhead L, Prat C et al (2006) Lentivirus-mediated gene transfer to the central nervous system: therapeutic and research applications. Hum Gene Ther 17:1–9PubMedCrossRefGoogle Scholar
  21. 21.
    David AL, Peebles D (2007) Gene therapy for the fetus: is there a future? Best Pract Res Clin Obstet Gynaecol 22:203–218PubMedCrossRefGoogle Scholar
  22. 22.
    Barbera A, Jones OW, Zerbe GW et al (1995) Early ultrasonographic detection of fetal growth retardation in an ovine model of placental insufficiency. Am J Obstet Gynecol 173:1071–1074PubMedCrossRefGoogle Scholar
  23. 23.
    Kelly RW, Newnham JP (1989) Estimation of gestational age in Merino ewes by ultrasound measurement of fetal head size. Aust J Agr Res 40:1293–1299CrossRefGoogle Scholar
  24. 24.
    Carr DJ, Aitken RP, Milne JS et al (2011) Ultrasonographic assessment of growth and estimation of birthweight in late gestation fetal sheep. Ultrasound Med Biol 37:1588–1595Google Scholar
  25. 25.
    Jeanty P, Dramaix-Wilmet M, Elkhazen N et al (1982) Measurements of fetal kidney growth on ultrasound. Radiology 144:159–162PubMedGoogle Scholar
  26. 26.
    Abi-Nader K, Mehta V, Wigley V et al (2009) Doppler ultrasonography for the noninvasive measurement of uterine artery volume blood flow through gestation in the pregnant sheep. Reprod Sci 17:13–19PubMedCrossRefGoogle Scholar
  27. 27.
    Newnham JP, Kelly RW, Boyne P, Reid SE (1989) Ultrasound guided blood sampling from fetal sheep. Aust J Agr Res 40:401–407CrossRefGoogle Scholar
  28. 28.
    Henderson DC (1990) The veterinary book for sheep farmers. Farming Press, IpswichGoogle Scholar
  29. 29.
    Wallace JM, Bourke DA, Aitken RP et al (2002) Blood flows and nutrient uptakes in growth-restricted pregnancies induced by overnourishing adolescent sheep. Am J Physiol Regul Integr Comp Physiol 282:R1027–R1036PubMedGoogle Scholar
  30. 30.
    Macrae JC, Bruce LA, Hovell DFD et al (1991) Influence of protein nutrition on the response of growing lambs to exogenous bovine growth hormone. J Endocrinol 130:53–61PubMedCrossRefGoogle Scholar
  31. 31.
    Dickerson KS, Newhouse VL, Tortoli P, Guidi G (1993) Comparison of conventional and transverse Doppler sonograms. J Ultrasound Med 12:497–506PubMedGoogle Scholar
  32. 32.
    Abi-Nader K, Mehta V, Shaw SWS et al (2010) Telemetric monitoring of fetal blood pressure and heart rate in the freely moving pregnant sheep: A feasibility study. Lab Anim Sci 45:50–54CrossRefGoogle Scholar
  33. 33.
    Wallace JM, Luther JS, Milne JS et al (2006) Nutritional modulation of adolescent pregnancy outcome – a review. Placenta 27(Suppl A):S61–S68PubMedCrossRefGoogle Scholar
  34. 34.
    Logan EF, Foster WH, Irwin D (1978) A note on bovine colostrum as an alternative source of immunoglobulins for lambs. Anim Prod 26:93–96CrossRefGoogle Scholar
  35. 35.
    Jubb KVF, Kennedy PC, Palmer N (1993) Pathology of domestic animals. Academic, LondonGoogle Scholar
  36. 36.
    Meyer DJ, Harvey JW (1998) (Meyer DJ and Harvey JW, Eds.) Veterinary laboratory medicine – interpretation and diagnosis. WB Saunders Company, Philadelphia, pp 157–186Google Scholar
  37. 37.
    Kaneko JJ, Harvey JW, Bruss M (1997) Clinical biochemistry of domestic animals. Academic, San DiegoGoogle Scholar
  38. 38.
    West HJ (1987) Changes in the concentrations of bile acids in the plasma of sheep with liver damage. Res Vet Sci 43:243–248PubMedGoogle Scholar
  39. 39.
    Anwer MS, Engelking LR, Gronwall R, Klentz RD (1976) Plasma bile acid elevation following carbon tetrachloride induced liver damage in dogs, sheep, calves and ponies. Res Vet Sci 20:127–130PubMedGoogle Scholar
  40. 40.
    Sutherland RJ, Deol HS, Hood PJ (1992) Changes in plasma bile acids, plasma amino acids, and hepatic enzyme pools as indices of functional impairment in liver-damaged sheep. Vet Clin Pathol 21:51–55PubMedCrossRefGoogle Scholar
  41. 41.
    Hardy KJ, Hoffman NE, Mihaly G et al (1980) Bile acid metabolism in fetal sheep; perinatal changes in the bile acid pool. J Physiol 309:1–11PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2012

Authors and Affiliations

  • Vedanta Mehta
    • 1
  • Khalil N. Abi-Nader
    • 1
  • David Carr
    • 1
    • 2
  • Jacqueline Wallace
    • 2
  • Charles Coutelle
    • 3
  • Simon N. Waddington
    • 4
  • Donald Peebles
    • 1
    Email author
  • Anna L. David
    • 1
  1. 1.Prenatal Cell and Gene Therapy Group, EGA Institute for Women’s HealthUniversity College LondonLondonUK
  2. 2.The Rowett Institute of Nutrition and HealthUniversity of AberdeenAberdeenUK
  3. 3.National Heart and Lung Institute, Molecular and Cellular Medicine SectionImperial College LondonLondonUK
  4. 4.Institute for Women’s Health, Gene Transfer Technology GroupUniversity College LondonLondonUK

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