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Paul Terasaki

Paul Terasaki. GENE THERAPY. Transplantation of the Pancreas, Liver, Bone Marrow and Islets are all procedures in which new functioning genes are transferred from one individual to another as treatments. Growing a New Pancreas?. We have all done it!. But can it be possible to

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Paul Terasaki

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  1. Paul Terasaki

  2. GENE THERAPY Transplantation of the Pancreas, Liver, Bone Marrow and Islets are all procedures in which new functioning genes are transferred from one individual to another as treatments

  3. GrowingaNew Pancreas?

  4. We have all done it! But can it be possible to repeat this miracle of nature in the laboratory?

  5. Progenitor Cells Are not fully differentiated Committed to a special tissue, e.g. pancreas

  6. Human early foetal pancreas precursor tissue under renal capsule NOD/scid mice After 6 months beta cell increase x 5000 Castaing et al Diabetologia (2001)

  7. There are Three Golden Rules for Making Surrogate -Cells

  8. Unfortunately, We Don’t Know Any of Them

  9. Shinya Yamanaka

  10. VASCULARIZEDFUNCTIONAL HUMAN LIVER from iSPC derived ORGAN BUD TRANSPLANT (Takebe and Taniguchi Nature 2013)

  11. iPSCells from Type 1diabetics differentiatedto produce Insulin PNAS 2009 Maehr et al. Melton’s Lab

  12. Caution iPS Cells can become antigenic and undergo rejection (Zhao et al Nature 2011 vol 474) Can accumulate DNA Abnormalaties (Christine Mummary NEJM 2011) Can Retain epigenitic memory of to Cell type of Origin (Kim et al Nature 2010

  13. Expression of desired gene • Coax with Differentiation factors • Engineer by gene insertion • Combined coaxing and engineering

  14. Manet

  15. Caravaggio

  16. Goya Goya

  17. Stem Cell Differentiating Cell “SOUP” Hypothesis Stem cells in a soup of growth & differentiation factors

  18. Genetic Engineering Electroporation Viral vectors Adeno & adeno-associated virus Retrovirus e.g. Lenti modified HIV can unmask oncogenes

  19. Autologus Cell Transplant No immunosuppressive drugs Requires suitable source of cells ? Autoimmune recurrence risk unless target eliminated

  20. In Vivo v Ex Vivo In Vivo - Reduced hazards of infection, but transfection difficulties Ex Vivo - Problem of where and how to transplant manipulated cells

  21. “TOO GOOD TO BE TRUE !” CURIOUS !! If it can be repeated in another laboratory, it would be interesting. Peter Medawar

  22. The Literature Crescendo Negotiating difficult and hazardous published traps

  23. Human Insulin Gene transfection of Autologous Hepatocytes Glucose Response Promoter EGR1 Plasmid ex-vivo electroporation Direct intra-hepatic cell injection Cured diabetic Pigs for up to 9 months (Singapore, Nelson, Oi Lian Kon et al 2008 +1)

  24. Vast literature scatteredin ever increasing numbers of journals ASSESSMENT -; 1) TRUE and INTERESTING 2) TRUE and of NO interest 3) FALSE due to experimental or perceptual error 4) FALSE due to FRAUD 3 and 4 confuse, lead others astray and can waste years of effort

  25. To show that “A” is true, you don’t do “B”, you do “A” again (Brian Nosek,quoted Nature 2012 Ed Yong) Unfortunately doing “A” again is BORING so replication experiments are UNPOPULAR

  26. Mohamed Ghoneim Mansoura Egypt

  27. Autologous Mesenchymal Stromal Cells (Mansoura) 1)Bone Marrow, Studies with MSC’s from rat to rat isologous (Gabr et al, exp. and clin. Transplantation, 2008) 2 HUMAN Bone Marrow MSC’s to diabetic scid mice,(Gabr et al, Cell Transplantation, 2012)

  28. Mesenchymalal Stromal cells

  29. Undifferentiated passage 3 un un

  30. Islet like clusters after 17 days culture

  31. Islet like clusters Ditizone after 17 days culture

  32. Insulin +ve (GREEN) & glucagon +ve (RED) differentiated MSCs transplanted under renal capsule (63X/1.40 OIL, Zoom factor 2.5)

  33. Insulin +ve (GREEN) differentiated MSCs transplanted under renal capsule (63X/1.40 OIL, Zoom Factor 2.5)

  34. Cpp +ve (RED) differentiated MSCs transplanted under renal capsule (63X/1.40 OIL, Zoom Factor 2.5)

  35. Insulin & Cpp +ve (Yellow) differentiated MSCs transplanted under renal capsule (63X/1.40 OIL, Zoom Factor 2.5)

  36. Insulin +ve (GREEN) differentiated MSCs transplanted under renal capsule (63X/1.40 OIL)

  37. Mansoura Culture Proliferation and Differentiation Small numbers of insulin producing cells 1-5% Probably Glucose Responsive 7 Days After transplantation insulin producing cells rise to 15%

  38. Data suggest Human bone marrow MSC’s can produce human insulin and C-peptide for at least 3 months in a xenogenic model Although small in amount there is sufficient to control ATZ diabetes in the diabetic SCID mouse

  39. Outstanding Problems Sufficient cells to provide adequate therapy Persistent gene expression & protein synthesis for a long period

  40. Encouraging in- vivo studies with the AAV Vector IM injections of the human insulin and Glucokinase genes into STZ diabetic rats (Mas et al, Diabetes 2006) and STZ diabetic dogs prolonged cure (Callejas et al Diabetes 2013) IV injection of factor IX in human Hemophilia B (Amit et al, NEJM 2011)

  41. iPS Cells and Mesenchymal Stromal Cells only new runners in 10 years

  42. BELIEVE NOTHING ! no matter where you read it or who has said it, not even if I have said it, unless it agrees with your own reason and your own common sense. Buddha

  43. Insulin And Cpp in cultutred differentiated MSCs using Trichostatintransplanted under renal capsule, received on 28- 8-2013

  44. Insulin +ve (GREEN) differentiated MSCs transplanted under renal capsule (63X/1.40 OIL)

  45. Cpp +ve (RED) differentiated MSCs transplanted under renal capsule (63X/1.40 OIL)

  46. Insulin & Cpp +ve (Yellow) differentiated MSCs transplanted under renal capsule (63X/1.40 OIL)

  47. Insulin +ve (GREEN) differentiated MSCs transplanted under renal capsule (63X/1.40 OIL, Zoom Factor 3)

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