Stem Cell Research and Cloning

1. Stem Cell Research and Cloning Doing Science in a Pluralistic Society David Lemberg, M.S., D.C. Associate Faculty National University

2. David Lemberg, M.S., D.C. Associate Faculty National University

3. “There are many different ends that men may seek and still be fully rational, fully men, capable of understanding each other and sympathizing and deriving light from each other, . . . worlds, outlooks, very remote from our own.” — Isaiah Berlin, The Pursuit of the Ideal “Yet with how many things are we upon the brink of becoming acquainted, if cowardice or carelessness did not restrain our inquiries.” — Mary Shelley, Frankenstein “Questions of use of science and technology are always moral and political questions, never simply technical ones.” — Leon Kass, The New Biology: What Price Relieving Man’s Estate? David Lemberg, M.S., D.C. Associate Faculty National University

4. I’m not a mistake, I’m not a fake, It’s set in my DNA —Miley Cyrus “Can’t Be Tamed” David Lemberg, M.S., D.C. Associate Faculty National University

5. David Lemberg, M.S., D.C. Associate Faculty National University

6. David Lemberg, M.S., D.C. Associate Faculty National University

7. David Lemberg, M.S., D.C. Associate Faculty National University

8. Dolly and the Cloning Aftermath • Birth of Dolly —announced February 1997 • Cloning of humans was universally deemed imminent • “The cloning of humans would fit precisely into Adolph Hitler’s world view” • —Die Welt (2-27-97) • Cloning of human beings would permit “a eugenic and racist selection of the human race” • —European Parliament (3-13-97) David Lemberg, M.S., D.C. Associate Faculty National University

9. History of Cloning • In 1903 plant physiologist Herbert John Webber described clons • Groups of plants that are propagated by the use of any form of vegetative parts such as bulbs, tubers, cuttings, . . . which are simply parts of the same individual” • Clon(e) is derived from the Greek klonmeaning “trunk” or “branch” David Lemberg, M.S., D.C. Associate Faculty National University

10. History of Cloning • In 1994 cloning was described as “taking the nucleus of a cell from the body of an adult and transferring it to an unfertilized egg, destroying the genome of the oocyte of the egg, and letting it develop”* • Two years before the birth of Dolly *Voelker R: A clone by any other name is still an ethical concern. JAMA 271:331–332, 1994 David Lemberg, M.S., D.C. Associate Faculty National University

11. History of Cloning • In 1894 Jacques Loeb cleaved fertilized sea urchin eggs • In some experiments two entire embryos would develop from the original zygotic material David Lemberg, M.S., D.C. Associate Faculty National University

12. History of Cloning • In 1914 Hans Spemann pinched off a portion of cytoplasm in salamander zygotes • If a nucleus migrated to the cytoplasm from one of the blastocyst cells, two embryos would result David Lemberg, M.S., D.C. Associate Faculty National University

13. Somatic Cell Nuclear Transfer • The origins of SCNT date from the 1950s and 1960s • Experiments on leopard frogs (Ranapipiens) • In a notable understatement, Robert Briggs and Thomas King declared “although the method of nuclear transplantation should be valuable principally for the study of nuclear differentiation, it may also have other uses” David Lemberg, M.S., D.C. Associate Faculty National University

14. David Lemberg, M.S., D.C. Associate Faculty National University

15. Cloning Mammals The next stage would be to clone mammals 1983 — James McGrath and DavorSolter of the Wistar Institute in Philadelphia utilized SCNT to clone mice In 1993 four calves were cloned from cultured cells derived from the inner cell mass of blastocysts David Lemberg, M.S., D.C. Associate Faculty National University

16. Cloning Mammals • Dolly’s birth was a big breakthrough — cloning a mammal from an adult somatic cell • Before Dolly, in October 1993, Jerry Hall created cloned human embryos by fission of a single human embryo • This method was a common technique in animal husbandry David Lemberg, M.S., D.C. Associate Faculty National University

17. David Lemberg, M.S., D.C. Associate Faculty National University

18. Cloning Mammals Robert Stillman, the director of the IVF program at George Washington University said the purpose was to help infertile couples who have “just the basic human desire to have a family” This is exactly a primary justification to proceed with research into human cloning David Lemberg, M.S., D.C. Associate Faculty National University

19. Cloning Mammals Very interestingly, Robert McKinnell at the University of Minnesota stated “it was much harder to take a cell from an adult organism and use it to make embryos or even clones, since the cells of adult organisms are committed to specific functions and have switched off their capacity for full development” Both of these hurdles have now been stunningly overcome David Lemberg, M.S., D.C. Associate Faculty National University

20. Cloning Mammals • The outcry against Hall’s work was immediate • George Annas—bioethicist • “This is the experiment we were never going to do— it’s a horror story” • Others suggested the public is “uncomfortable with meddling with the life-reproducing process” David Lemberg, M.S., D.C. Associate Faculty National University

21. Cloning Mammals The Vatican’s official newspaper, L’Osservatore Romano, stated the U.S. must regulate “unscrupulous scientists who venture into a tunnel of madness” In contrast, medical ethicist Norman Fost said these decisions should be the parents’ prerogative — “a presumption of privacy and liberty, that people should be able to live their lives the way they want and to make babies the way they want” David Lemberg, M.S., D.C. Associate Faculty National University

22. Dolly and the Cloning Aftermath • Birth of Dolly —announced February 1997 • Cloning of humans was universally deemed imminent • “The cloning of humans would fit precisely into Adolph Hitler’s world view” • —Die Welt (2-27-97) • Cloning of human beings would permit “a eugenic and racist selection of the human race” • —European Parliament (3-13-97) David Lemberg, M.S., D.C. Associate Faculty National University

23. David Lemberg, M.S., D.C. Associate Faculty National University

24. Somatic Cell Nuclear Transfer • Wilmut’s team utilized SCNT • Dolly derived from a clone of an adult somatic cell • These cell populations were clones of mammary gland cells of a 6-year-old ewe David Lemberg, M.S., D.C. Associate Faculty National University

25. Stem Cell Research Remarkably the report of the isolation and growth of human stem cells soon followed the announcement of the birth of Dolly James Thomson’s team at the University of Wisconsin cultured 5 independent cell lines from the ICMs of 14 blastocysts David Lemberg, M.S., D.C. Associate Faculty National University

26. Thomson Lab Stem Cells Fig. 1. Derivation of the H9 cell line. (A) Inner cell mass-derived cells attached to mouse embryonic fibroblast feeder layer after 8 days of culture, 24 hours before first dissociation. Scale bar, 100 mm. (B) H9 colony. Scale bar, 100 mm. (C) H9 cells. Scale bar, 50 mm. (D) Differentiated H9 cells, cultured for 5 days in the absence of mouse embryonic fibroblasts, but in the presence of human LIF (20 ng/ml; Sigma). Scale bar, 100 mm. Science 282(5391):1145-1147, 1998 Human Leukemia Inhibitory Factor (LIF) is a lymphoid factor that promotes long-term maintenance of embryonic stem cells by suppressing spontaneous differentiation. David Lemberg, M.S., D.C. Associate Faculty National University

27. Stem Cell Research Thomson’s paper was titled “Embryonic Stem Cell Lines Derived from Human Blastocysts” The article concluded “progress in developmental biology is now extremely rapid. Human ESCs will link this progress even more closely to the prevention and treatment of human disease.” David Lemberg, M.S., D.C. Associate Faculty National University

28. David Lemberg, M.S., D.C. Associate Faculty National University

29. Stem Cell Research In August 2001, the same month Time placed Thomson on its cover, President Bush severely restricted federal funding for stem cell research Bush declared “Research on embryonic stem cells raises profound ethical questions, because extracting the stem cells destroys the embryo and thus destroys its potential for life.” David Lemberg, M.S., D.C. Associate Faculty National University

30. SC Research — Ethical Issues • Until two years ago, stem cells were exclusively derived from an embryo • From the inner cell mass of a blastocyst David Lemberg, M.S., D.C. Associate Faculty National University

31. David Lemberg, M.S., D.C. Associate Faculty National University

32. David Lemberg, M.S., D.C. Associate Faculty National University

33. SC Research — Ethical Issues Isolating stem cells from the ICM kills the embryo Many institutions and organizations believe this is equivalent to killing a born human being David Lemberg, M.S., D.C. Associate Faculty National University

34. SC Research — Ethical Issues • Richard Doerflinger • Associate Director of the U.S. Conference of Catholic Bishops' Secretariat of Pro-Life Activities • Doerflinger states that embryo research “directly promotes the destruction of embryonic human life” and destruction of embryos represents “wrongful killing” • Doerflinger RM: The ethics of funding embryonic stem cell research. A Catholic viewpoint. Kennedy Inst Ethics J 9(2):137-150, 1999 • Stem cells are “lethally harvested from embryos” David Lemberg, M.S., D.C. Associate Faculty National University

35. SC Research — Ethical Issues • The Catholic church, as a monolithic entity, is opposed to all extracorporeal activities involving embryos. • The Vatican’s Instruction Dignitas Personae advises the faithful — • “The fruit of human generation, from the first moment of its existence, that is to say, from the moment the zygote has formed, demands the unconditional respect that is morally due to the human being in his bodily and spiritual totality” • Vatican: Congregation for the Doctrine of the Faith. Instruction Dignitas Personae on Certain Bioethical Questions, September 8, 2008 David Lemberg, M.S., D.C. Associate Faculty National University

36. SC Research — Ethical Issues • Dignitas Personae • Experimentation on human embryos “constitutes a crime against their dignity as human beings . . . [and] always constitutes a grave moral disorder” • “The use of embryonic stem cells . . . even when these are provided by other researchers through the destruction of embryos . . . presents serious problems from the standpoint of cooperation in evil and scandal” David Lemberg, M.S., D.C. Associate Faculty National University

37. Moral Status of the Embryo • On the previous views, the embryo is a human being with full moral status • Embryo have all the rights and privileges that born human beings enjoy • The opposite view holds that the embryo has no moral status whatsoever • A middle ground may be sought in which the embryo holds no moral status but yet is entitled to moral respect David Lemberg, M.S., D.C. Associate Faculty National University

38. Moral Status of the Embryo • As moral agents we acknowledge the moral value of embryos and accord them respect • We do embryo research to fulfill specific, meaningful, high-impact goals • Improving assisted reproductive technologies • Improving contraception • Finding solutions to genetic diseases • But of course many remain vigorously opposed to such research David Lemberg, M.S., D.C. Associate Faculty National University

39. Sources of Embryos for Research • Creating embryos specifically for research • This is highly contentious • In his August 2001 Presidential Statement, President Bush described the “deeply troubling” activity of creating embryos “solely to experiment on them” David Lemberg, M.S., D.C. Associate Faculty National University

40. Sources of Embryos for Research • Instrumentalism — created human embryos are intended to be used as a means only — the ends of the embryos are not considered • Immanuel Kant’s Formula of Humanity as an End in Itself — “So act that you use humanity . . . always at the same time as an end, never merely as a means” • Wood AW: Kant. Oxford, UK, Blackwell, 2005, p 135 • But — embryos are not sentient, they have no interests, and they have no ends David Lemberg, M.S., D.C. Associate Faculty National University

41. Sources of Embryos for Research Use of “spare” embryos Spare embryos are produced in hyperovulation cycles and unused in IVF procedures Spare embryos are contrasted with embryos created specifically for research Thus, the spare–created distinction David Lemberg, M.S., D.C. Associate Faculty National University

42. Sources of Embryos for Research • In IVF, spare embryos are either frozen for later use or destroyed • Using these embryos for research represents a third alternative • But spare embryos are often not of top quality • Many have various defects that caused them to be eliminated from the pool of candidates for implantation • Others have been stored for many years David Lemberg, M.S., D.C. Associate Faculty National University

43. Sources of Embryos for Research • Research based on these [spare] embryos can lead to seriously misleading conclusions* • *Green RM: The Human Embryo Research Debates. New York, Oxford University Press, 2001, p 15 David Lemberg, M.S., D.C. Associate Faculty National University

44. Stem Cell Research — The Way Forward? How to make federal choices in a pluralistic society? Conflicts between personal autonomy and compelling state interests Need for long overdue system of ethical review and oversight Need for careful study, deliberation, and guidelines for the conduct of both research and clinical practice David Lemberg, M.S., D.C. Associate Faculty National University

45. Inappropriate SC Applications • “Trend toward transplants in which the stem cells are expected to behave in different ways, even though there is little empirical evidence to suggest they can do so” • Report in June 2010 of the “discovery of strange lumps of cells in the kidney of a woman who had undergone stem-cell treatment in Thailand” • Nature 465:997, 2010 David Lemberg, M.S., D.C. Associate Faculty National University

46. Inappropriate SC Applications • In Russia, a boy with ataxia telangiectasia (Louis-Bar syndrome; degeneration of the cerebellum, with dyscoordinated movement and speech) wasinjected with fetal neural stem cells • Four years later he developed multifocal tumors in the brain stem and cauda equine • The tumors were derived from the injected fetal cells David Lemberg, M.S., D.C. Associate Faculty National University

47. Inappropriate SC Applications • 200 clinics worldwide – 100 in China alone – offering unproven SC treatments for scores of disorders including MS, ALS, and spinal cord injury • Nature 459:147, 2009 • The potential profits are huge • Few facilities offer evidence from controlled clinical trials or from rigorous follow-up of patients David Lemberg, M.S., D.C. Associate Faculty National University

48. Induced Pluripotent Stem Cells • iPSCs may substitute for embryonic stem cells • In 2007 two teams simultaneously announced the creation of iPSCs • Yamanaka et al at Kyoto University* • Thomson et al at the University of Wisconsin** *Takahashi K, Yamanaka S: Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126(4):663–676, 2006 **Yu J, et al: Induced pluripotent stem cell lines derived from human somatic cells. Science 318(5858):1917‑1920, 2007. David Lemberg, M.S., D.C. Associate Faculty National University

49. Induced Pluripotent Stem Cells • The Yamanaka team utilized four transcription factors to reprogram adult differentiated cells to a pluripotent state • OCT4, SOX2, KLF4, and c-MYC • This work was groundbreaking • The primary obstacle going forward was the use of a retroviral vector to deliver the transcription factors David Lemberg, M.S., D.C. Associate Faculty National University

50. Induced Pluripotent Stem Cells Importantly, the creation of iPSCs does not involve the destruction of embryos The White House said that President Bush was “very pleased” about the new findings, adding that “By avoiding techniques that destroy life, while vigorously supporting alternative approaches, President Bush is encouraging scientific advancement within ethical boundaries.” David Lemberg, M.S., D.C. Associate Faculty National University