Article contents
Germ-Line Therapy to Cure Mitochondrial Disease: Protocol and Ethics of In Vitro Ovum Nuclear Transplantation
Published online by Cambridge University Press: 29 July 2009
Extract
The combination of genuine ethical concerns and fear of learning to use germ-line therapy for human disease must now be confronted. Until now, no established techniques were available to perform this treatment on a human. Through an integration of several fields of science and medicine, we have developed a nine step protocol at the germ-line level for the curative treatment of a genetic disease. Our purpose in this paper is to provide the first method to apply germ-line therapy to treat those not yet born, who are destined to have a life threatening, or a severely debilitating genetic disease. We hope this proposal will initiate the process of a thorough analysis from both the scientific and ethical communities. As such, this proposal can be useful for official groups studying the advantages and disadvantages of germ-line therapy.
- Type
- Special Section: Designs on Life: Choice, Control, and Responsibility in Genetic Manipulation
- Information
- Copyright
- Copyright © Cambridge University Press 1995
References
Notes
1. President's Commission for the Study of Ethical Problems in Medicine and Biomedical and Behavioral Research. Splicing Life. Washington, DC: Government Printing Office, 1982.Google Scholar
2. Wivel, NA, Walters, L. Germ-line gene modification and disease prevention: some medical and ethical perspectives. Science 1993;262:533–8.CrossRefGoogle ScholarPubMed
3. de Graf, J, Swinkels, DW, de Haan, AF, Demacker, PN, Stalenhoef, AF. Both inherited susceptibility and environmental exposure determine the low density lipoprotein-subfraction pattern distribution in healthy Dutch families. American Journal of Human Genetics 1992;51:1295–1310.Google Scholar
4. Stern, HP, Gonzalez, C, Mitchell, BD, Villalpando, E, Haffner, SM, Hazuda, HP. Genetic and environmental determinants of type II diabetes in Mexico City and San Antonio. Diabetes 1992;41:484–92.CrossRefGoogle ScholarPubMed
5. Arshad, SH, Hide, DW. Effect of environmental factors on the development of allergenic disorders in infancy. Journal of Allergy and Clinical Immunology 1992;90:235–241.CrossRefGoogle Scholar
6. Fletcher, RH, Fletcher, SW. Teaching preventive medicine and health maintenance. Annals of Internal Medicine 1992;116:1094–8.CrossRefGoogle ScholarPubMed
7. Collins, TR, Goldenberg, K, Ring, A, Nelson, K, Konen, J. The Association of Teachers of Preventive Medicine's recommendation for postgraduate education in prevention. Academic Medicine 1991;66:317–20.CrossRefGoogle Scholar
8. Anderson, WF. Human gene therapy. Science 1992;256:808–13.CrossRefGoogle ScholarPubMed
9. Karson, EM, Polvino, W, Anderson, WF. Prospects for human gene therapy. Journal of Reproductive Medicine 1992;37:508–14.Google ScholarPubMed
10. Wallace, DC. Diseases of the mitochondrial DNA. Annual Review of Biochemistry 1992;61:1174–212.CrossRefGoogle ScholarPubMed
11. .Gyllensten, U, Whartin, D, Joseffson, A, Wilson, AC. Paternal inheritance of mitochondrial DNA in mice. Nature 1991;352:255–7.CrossRefGoogle ScholarPubMed
12. Walters, L. The ethics of human gene therapy. Nature 1986;320:225–7.CrossRefGoogle ScholarPubMed
13. Office of Technology Assessment. New developments in biotechnology-background paper: public perceptions of biotechnology, Washington, DC: US Government Printing Office, 1987.Google Scholar
14. Walters, L. Human gene therapy: ethics and public policy. Human Gene Therapy 1991;2:115–22.CrossRefGoogle ScholarPubMed
15. Pavlakis, SG, Phillips, PC, DiMauro, S, Devivo, DC, Rowland, LP. Mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes: a distinctive clinical syndrome. Annals of Neurology 1984;16:481–7.CrossRefGoogle ScholarPubMed
16. Goto, Y-I, Nonaka, I, Horai, S. A mutation in the tRNALeu(UUR) gene associated with the MELAS subgroup of mitochondrial encephalomyopathies. Nature 1990;348:651–3.CrossRefGoogle ScholarPubMed
17. Hirano, M, Ricci, E, Koenigsberger, MR et al. , MELAS: an original case and clinical criteria for diagnosis. Neuromuscular Disorders 1992;2:125–35.CrossRefGoogle ScholarPubMed
18. MEDLINE [database online]. Bethesda (MD): National Library of Medicine, 1989, updated weekly. Available from: National Library of Medicine; BRS Information Technologies, McLean, VA; DIALOG Information Services, Inc., Palo Alto, CA; et al. 1994.Google Scholar
19. Anderson, S, Bankier, AT, Barrell, BG et al. , Sequence and organization of the human mitochondrial genome. Nature 1981;290:457–65.CrossRefGoogle ScholarPubMed
20. Satoh, M, Kutoiwa, T. Organization of multiple nucleoids and DNA molecules in mitochondria of a human cell. Experimental Cell Research 1991;196:137–40.CrossRefGoogle ScholarPubMed
21. Piko, L, Matsumoto, L. Number of mitochondria and some properties of mitochondria DNA in the mouse egg. Developmental Biology 1976;49:1–10.CrossRefGoogle ScholarPubMed
22. Piko, L, Hougham, AJ, Bulpitt, KJ. Studies of sequence heterogeneity of mitochondria' DNA from rat and mouse tissues; evidence for an increased frequency of deletions/additions with aging. Mechanisms of Ageing Development 1988;43:279–93.CrossRefGoogle Scholar
23. Michaels, GS, Hauswirth, WW, Laipis, PJ. Mitochondria' DNA copy number in bovine oocytes and somatic cells. Developmental Biology 1982;94:246-51.CrossRefGoogle Scholar
24. Schon, EA. Mitochondria' disorders in muscle. Current Opinions in Neurology & Neurosurgery 1992;6:19–26.Google Scholar
25. Clayton, DA, Dodsa, JN, Friedberg, EC. The absence of a pyrimidine dimer repair mechanism in mammalian mitochondria. Proceedings of the National Academy of Sciences USA 1974;71:2777–81.CrossRefGoogle ScholarPubMed
26. Richter, C, Park, J-W, Ames, BN. Normal oxidative damage to mitochondria' and nuclear DNA is extensive. Proceedings of the National Academy of Sciences USA 1988;85:6465–7.CrossRefGoogle Scholar
27. Pettepher, CC, LeDoux, SP, Bohr, VA, Wilson, GL. Repair of alkali-labile sites within the mitochondria' DNA of RIN1r 38 cells after exposure to the nitrosurea streptozotocin. Journal of Biological Chemistry 1991;266:3113–7.Google Scholar
28. Holt, IJ, Harding, AE, Petty, RKH, Morgan-Hughes, JA. A new mitochondria' disease associated with mitochondrial DNA heteroplasmy. American Journal of Human Genetics 1990;46:428–33.Google Scholar
29. Tatuch, Y, Christodolou, J, Feigenbaum, A et al. , Heteroplasmic mtDNA mutation (T→ G) at 8993 can cause Leigh's disease when the percentage of abnormal mtDNA is high. American Journal of Human Genetics 1992;50:852–8.Google Scholar
30. Shoffner, JM, Fernhoff, PM, Krawiecki, NS et al. , Subacute necrotizing encephalopathy: oxidative phosphorylation defects and the ATPase 6 point mutation. Neurology 1992;42:2168–74.CrossRefGoogle ScholarPubMed
31. Ciafaloni, E, Santorelli, FM, Shanske, S et al. , Maternally inherited Leigh syndrome. Journal of Pediatrics 1993;122:419–22.CrossRefGoogle ScholarPubMed
32. Shoffner, JM, Lott, MT, Lezza, AMS, Seibel, P, Ballinger, SW, Wallace, DC. Myoclonic epilepsy and ragged-red fiber disease (MERRF) is associated with a mitochondria' DNA tRNALYs mutation. Cell 1990;61:931–7.CrossRefGoogle Scholar
33. Silvestri, G, Moraes, CT, Shanske, S, Oh, STDiMauro, S. A new mtDNA mutation in the tRNALYs gene associated with myoclonic epilepsy and ragged-red fibers (MERRF). American Journal of Human Genetics 1992;51:1213–7.Google Scholar
34. Goto, Y-I, Nonaka, I, Horai, S. A new mIDNA mutation associated with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS). Biochimica Biophysica ACTA 1991;1097:238–40.CrossRefGoogle Scholar
35. van den Ouweland, JMW, Lemkes, HHPJ, Ruitenbeek, W et al. , Mutation in mitochondria' tRNALeU(UUR) gene in a large pedigree with maternally transmitted type II diabetes mellitus and deafness. Nature Genetics 1992;1:368–71.CrossRefGoogle Scholar
36. Moraes, CT, Ciacci, F, Silvestri, G et al. , Atypical clinical presentations with the MELAS mutation at position 3243 of human mitochondria' DNA. Neuromuscular Disorders 1993;3:43–50.CrossRefGoogle Scholar
37. Zeviani, M, Gellera, C, Antozzi, C et al. , Maternally inherited myopathy and cardiomyopathy: association with mutation in mitochondrial DNA tRNALeu(UUR). Lancet 1991;338:143–7.CrossRefGoogle Scholar
38. Goto, Y-I, Tojo, M, Tohyyama, J, Horai, S, Nonaka, I. A novel point mutation in the mitochondria' DNA tRNALeu(UUR) gene in a family with mitochondrial myopathy. Annals of Neurology 1992;31:672–5.CrossRefGoogle Scholar
39. Moraes, CT, Ciacci, F, Bonilla, E, lonasecu, V, Schon, EA, DiMauro, S. A mitochondrial tRNA anticodon swap associated with a muscle disease. Nature Genetics 1993;4:284–8.CrossRefGoogle ScholarPubMed
40. Bindoff, LA, Howell, N, Poulton, J et al. , Abnormal RNA processing associated with a novel tRNA mutation in mitochondria' DNA. Journal of Biological Chemistry 1993;268:19559–64.Google Scholar
41. Moraes, CT, Ciacci, F, Bonilla, E et al. , Two novel pathogenic mitochondria' DNA mutations affect-Le ing organelle number and protein synthesis. Is the tRNAu(uut6) gene an etiological hotspot? Journal of Clinical Investigations 1993;92:2906.CrossRefGoogle Scholar
42. Taniike, M, Fukushima, H, Yangihar, I et al. , Mitochondria' tRNAlie mutation in fatal cardiomyopathy. Biochimica Biophysica ACTA 1992;186:47–53.Google Scholar
43. Yoon, KL, Aprille, JR, Ernst, SG. Mitochondria' tRNAThr mutation in fatal infantile respiratory enzyme deficiency. Biochemical and Biophysical Research Communications 1991;176:1112–5.CrossRefGoogle Scholar
44. Prezant, TR, Agapian, JV, Pohlman, MC et al. , Mitochondrial ribosomal RNA mutation associated with both antibiotic-induced and non-syndromic deafness. Nature Genetics 1993;4:289–94.CrossRefGoogle ScholarPubMed
45. Zeviani, M, Moraes, CT, DiMauro, S et al. , Deletions of mitochondrial DNA in Kearns-Sayre syndrome. Neurology 1988;38:1339–46.CrossRefGoogle ScholarPubMed
46. Poulton, J, Deadman, ME, Gardiner, RM. Duplications of mitochondria' DNA in mitochondrial myopathy. Lancet 1993;1:236–40.Google Scholar
47. Moraes, CT, DiMauro, S, Zeviani, M et al. , Mitochondrial DNA deletions in progressive external ophthalmoplegia and Kearns-Sayre syndrome. New England Journal of Medicine 1989;320:1293–9.CrossRefGoogle ScholarPubMed
48. Rotig, A, Cormier, V, Koll, F et al. , Site-specific deletions of the mitochondrial genome in the Pearson marrow-pancreas syndrome. Genomics 1991;10:502–4.CrossRefGoogle ScholarPubMed
49. Rotig, A, Bessis, J-L, Romero, N et al. , Maternally inherited duplication of the mitochondria' genome in a syndrome of proximal tubulopathy, diabetes mellitus, and cerebellar ataxia. American Journal of Human Genetics 1992;50:364–70.Google Scholar
50. Bernes, SM, Bacino, C, Prezant, TR et al. , Identical mitochondrial DNA deletion in mother with progressive external ophthalmoplegia and son with Pearson marrow-pancreas syndrome. Journal of Pediatrics 1993;123:598–602.CrossRefGoogle ScholarPubMed
51. Larsson, N-G, Eiken, HG, Holme, E, Oldfors, A, Tulinius, MH. Lack of transmission of deleted mtDNA from a woman with Kearns-Sayre syndrome to her child. American Journal of Human Genetics 1990; 50:360–3.Google Scholar
52. Ballinger, SW, Shoffner, JM, Hedaya, EV et al. , Maternally transmitted diabetes and deafness associated with a 10.4kb mitochondrial deletion. Nature Genetics 1992;1:11–5.CrossRefGoogle Scholar
53. Dunbar, DR, Moonie, PA, Swingler, RJ, Davidson, D, Roberts, R, Holt, IJ. Maternally transmitted partial direct tandem duplication of mitochondrial DNA associated with diabetes mellitus. Human Molecular Genetics 1993;2:1619–24.CrossRefGoogle ScholarPubMed
54. Zeviani, M, Bresolin, N, Gellera, C et al. , Nucleus-driven multiple large scale deletions of the human mitochondrial genome: a new autosomal dominant disease. American Journal of Human Genetics 1990;47:904–14.Google ScholarPubMed
55. Moraes, CT, Shanske, S, Tritschler, H-J et al. , Mitochondrial DNA depletion with variable tissue expression: a novel genetic abnormality in mitochondrial diseases. American Journal of Human Genetics 1991;48:492–501.Google ScholarPubMed
56. Tritschler, H-J, Andreeta, F, Moraes, CT et al. , Mitochondrial myopathy of childhood associated with depletion of mitochondrial DNA. Neurology 1992;42:209–17.CrossRefGoogle ScholarPubMed
57. Ciafaloni, E, Ricci, E, Shanske, S et al. , MELAS: clinical features, biochemistry, and molecular genetics. Annals of Neurology 1992;31:391–8.CrossRefGoogle ScholarPubMed
58. Martinuzzi, A, Bartolomei, L, Carrozzo, R et al. , Correlation between clinical and molecular features in two MELAS families. Journal of Neurological Science 1992;113:222–9.CrossRefGoogle ScholarPubMed
59. Boursot, P, Yonekawa, H, Bonhomme, F. Heteroplasmy in mice with deletion of a large coding region of mitochondrial DNA. Molecular Biology of Evolution 1987;4:46–55.Google ScholarPubMed
60. King, MP, Attardi, G. Human cells lacking mtDNA: repopulation with exogenous mitochondria by complementation. Science 1989;246:–500.CrossRefGoogle Scholar
61. Chomyn, A, Meola, G, Bresolin, N, Lai, ST, Scarlato, G, Attardi, G. In vitro genetic transfer of protein synthesis and respiration defects to mitochondrial DNA-less cells with myopathy-patient mitochondria. Molecular and Cellular Biology 1991;11:2236–44.CrossRefGoogle ScholarPubMed
62. Chomyn, A, Martinuzzi, A, Yoneda, M et al. , MELAS mutation in mtDNA binding site for transcription termination factor causes defects in protein synthesis and in respiration but no changes in levels of upstream and downstream mature transcripts. Proceedings of the National Academy of Sciences USA 1992;89:4221–5.CrossRefGoogle Scholar
63. King, MP, Koga, Y, Davidson, M, Schon, EA. Defects in mitochondrial protein synthesis and respiratory chain activity segregate with the tRNALeu(uuR) mutation associated with MELAS. Molecular and Cellular Biology 1992;12:480–90.CrossRefGoogle Scholar
64. Hayashi, J-I, Ohta, S, Kikuchi, A, Takemitsu, M, Goto, Y-I, Nonaka, I. Introduction of disease-related mitochondrial DNA deletions into HeLa cells lacking mitochondria] DNA results in mitochondria] dysfunction. Proceedings of the National Academy of Sciences USA 1991;88:10614–8.CrossRefGoogle ScholarPubMed
65. McGrath, J, Solter, D. Nuclear transplantation in the mouse embryo by microsurgery and cell fusion. Science 1983;220:1300–2.CrossRefGoogle ScholarPubMed
66. Edirisinghe, WR, Wales, RG, Chapman, HM, Yovichi, JL. Assisted fertilization of mouse oocytes and preliminary results for human oocytes using zona drilling. Journal of In Vitro Fertilization and Embryo Transfer 1991;8:48–55.CrossRefGoogle ScholarPubMed
67. Cheong, H-T, Takahashi, Y, Kanagawa, H. Birth of mice after transplantation of early cell-cyclestage embryonic nuclei into enucleated oocytes. Biological Reproduction 1993;48:958–63.CrossRefGoogle ScholarPubMed
68. Kono, T, Sotomaru, Y, Sato, Y, Nakahara, T. Development of androgenetic mouse embryos produced by in vitro fertilization of enucleated oocytes. Molecular Reproduction and Development 1993;34:43–6.CrossRefGoogle ScholarPubMed
69. Yoneda, M, Chomyn, A, Martinuzzi, A, Hurko, O, Attardi, G. Marked replicative advantage of human mtDNA carrying a point mutation that causes the MELAS encephalomyopathy. Proceedings of the National Academy of Sciences USA 1992;89:–11164–8.CrossRefGoogle ScholarPubMed
70. Ponsa, M, Nogues, C, Vidal, F, Egozcue, J. Scanning electron microscope (SEM) study of mouse embryos obtained from isolated blastomeres. Journal of In Vitro Fertilization and Embryo Transfer 1991;8:279–85.CrossRefGoogle ScholarPubMed
71. Bonilla, E, Sciacco, M, Tanji, K, Sparaco, M, Petruzella, V, Moraes, CT. New morphological approaches to the study of mitochondrial encephalomyopathies. Brain Pathology 1992;2:113–9.CrossRefGoogle Scholar
72. Schon, EA, Rizzuto, R, Moraes, CT, Nakase, H, Zeviani, M, Dimauro, S. A direct repeat is a hotspot for large-scale deletions of human mitochondrial DNA. Science 1989;244:346–9.CrossRefGoogle ScholarPubMed
73. Chen, LB. Fluorescent labeling of mitochondria. Methods in Cell Biology 1989;29:103–23.CrossRefGoogle ScholarPubMed
74. Reers, M, Smith, TW, Chen, LB. J-aggregate formation of a carbocyanine as a quantitative fluorescent indicator of membrane potential. Biochemistry 1991;30:4480–6.CrossRefGoogle ScholarPubMed
75. Serhal, PH, Craft, IL. Ovum donation -a simplified approach. Fertility and Sterility 1987;48:265–9.CrossRefGoogle ScholarPubMed
76. Awadalla, SG, Friedman, CI, Chin, NW, Dodds, W, Park, J, Kim, MH. Follicular stimulation for in vitro fertilization using pituitary suppression and human menopausal gonadotropins. Fertility and Sterility 1987;48:811–5.CrossRefGoogle ScholarPubMed
77. lllmensee, K, Hoppe, PC. Nuclear transplantation in Mus musculus: developmental potential of nuclei from preimplantation embryos. Cell 1981;23:9–18.CrossRefGoogle Scholar
78. Robl, JM, First, NL. Manipulation of gametes and embryos in the pig. Journal of Reproduction and Fertility 1985;33:101–14.Google ScholarPubMed
79. Rob, JM, Prather, RS, Barnes, F et al. , Nuclear transplantation in bovine embryos. Journal of Animal Science 1987;64:642–7.Google Scholar
80. Prather, RS, First, NL. Cloning embryos by nuclear transfer. Journal of Reproduction and Fertility 1990;41:125–34.Google ScholarPubMed
81. Kono, T, Kwon, OY, Watanabe, T, Nakahara, T. Development of mouse enucleated oocytes receiving a nucleus from different stages of the second cell cycle. Journal of Reproduction and Fertility 1992;94:481–7.CrossRefGoogle ScholarPubMed
82. Whitaker, MJ. Cell cycle control proteins are second messenger targets at fertilization in sea-urchin eggs. Journal of Reproduction and Fertility 1990;42:199–204.Google ScholarPubMed
83. Hendyside, AL, Lesko, JG, Tarin, JJ, Winston, ML, Hughes, MR. Birth of a normal girl after in vitro fertilization and preimplantation diagnostic testing for cystic fibrosis. New England Journal of Medicine 1992;327:905–9.CrossRefGoogle Scholar
84. Rigger, EF, Bustillo, M, Dorfmann, AD, Schulman, JD. Human preimplantation embryo cryopreservation: selected aspects. Human Reproduction 1991;6:131–5.Google Scholar
85. Shanske, AL, Shanske, S, Silvestri, G, Tanji, K, Wertheim, D, Lipper, S. MELAS point mutation with unusual clinical presentation. Neuromuscular Disorders 1993.CrossRefGoogle ScholarPubMed
86. Cometta, K, Wieder, R, Anderson, WF. Gene transfer into primates and prospects for gene therapy in humans. Progression in Nucleic Acid Research and Molecular Biology 1989;36:311–22.Google Scholar
87. Roemer, K, Friedman, T. Concepts and strategies for human gene therapy. European Journal of Biochemistry 1992;208:211–25.CrossRefGoogle ScholarPubMed
88. van de Water, J, Gershwin, ME, Leung, P, Ansari, A, Coppel, RL. The autoepitope of the 74 kD mitochondrial autoantigen of primary biliary cirrhosis corresponds to the functional site of dihydrolipoamide acetyltransferase. Journal of Experimental Medicine 1988;167:1791–9.CrossRefGoogle ScholarPubMed
89. Schapira, AHVCooper, JM, Manneschi, L, Vital, C, Morgan-Hughes, JA, Clark, JB. A mitochondrial mwephalomyopathy with specific deficiencies of two respiratory chain polypeptides and a circulation autoantibody to a mitochondrial matrix protein. Brain 1993;113:419–32.CrossRefGoogle Scholar
90. Loveland, B, Wang, C-R, Yonekawa, H, Hermel, E, Fischer, Lindahl K. Maternally transmitted histocompatibility antigen of mice: a hydrophobic peptide of a mitochondrially encoded protein. Cell 1990;60:971–80.CrossRefGoogle ScholarPubMed
91. Dingwall, C, Laskey, R. The nuclear membrane. Science 1992;258:942–7.CrossRefGoogle ScholarPubMed
92. Leno, GH, Laskey, RA. The nuclear membrane determines the timing of DNA replication in Xenopus egg extracts. Journal of Cell Biology 1991;112:557–66.CrossRefGoogle ScholarPubMed
93. Congregation for the doctrine of the faith: respect for human embryos. In: Instruction on Respect for Human Life in Its Origin and on the Dignity of Procreation: Replies to Certain Questions of the Day. Vatican City: Vatican Polyglot Press, 1987:12–20.Google Scholar
94. Fletcher, J. The Ethics of Genetic Control. New York: Prometheus Books, 1988.Google Scholar
95. Seydel, FD, Human gene therapy-playing God? Progress in Clinical and Biological Research 1990; 352:339–346.Google ScholarPubMed
96. Temple, MJ. An ethical analysis of gene therapy as molecular surgery. The biology of hematopoiesis. Progress in Clinical and Biological Research 1990;352:347–53.Google Scholar
97. Anderson, WE. Human gene therapy: scientific and ethical considerations. Journal of Medical Philosophy 1985;10:275–92.CrossRefGoogle ScholarPubMed
98. Anderson, WF. Human gene therapy: why draw a line? Journal of Medical Philosophy 1989;14:681–93.CrossRefGoogle ScholarPubMed
99. Gray, MW. The endosymbiont hypothesis revisited. International Review of Cytology 1992;141:233–357.CrossRefGoogle ScholarPubMed
100. Wolstenholme, DR. Animal mitochondrial DNA: structure and evolution. International Review of Cytology 1992;141:173–216.CrossRefGoogle ScholarPubMed
101. Fletcher, J. Evolution of ethical debate about human gene therapy. Human Gene Therapy 1990; 1:55–68.CrossRefGoogle ScholarPubMed
- 38
- Cited by