1. In The Name Of God

2. Targeting Gene Therapy to Cancer

3. Abstract

4.  In recent years the idea of using gene therapy specially,cancer gene therapy will need to combine highly selective gene delivery with highly specific gene expression,specific gene product activity and specific drug activation.

5. This presentation will discuss the progress has made in recent years about the efficient delivery of DNA to tumour sites.

6. Introduction

7. MAJOR DEVELOPMENT IN GENE THERAPY

8. 1989:THE FIRST PROTOCOL FOR CANCER GENE THERAPY 1989:THE FIRST PROTOCOL FOR CANCER GENE THERAPY 1990:GENE THERAPY TO TREAT SCID USING STEM CELLS 1990:GENE THERAPY TO TREAT SCID USING STEM CELLS 1993:NEW GENE THERAPY APPROACH REPAIRS ERRORS IN mRNA 1993:NEW GENE THERAPY APPROACH REPAIRS ERRORS IN mRNA New approach using liposome 25 nanometers New approach using liposome 25 nanometers

9. SICKLE CELL DISEASE IS A SUCCESSFULLY TREATED IN MICE 2003:USING LIPOSOME COATED WITH PEG TO TREAT PARKINSON DISEASE USING siRNAs TO TREAT HUNTINGTON 2006:THE FIRST REPORT THAT GENE THERAPY CAN BE EFFECTIVE IN TREATING CANCER PREVENTION THE IMMUNE SYSTEM WITH Micro RNA 2007:THE FIRST GENE THERAPY TRIAL FOR INHERITED RETINAL DISEASE 2008:TREATMENT OF LEBER CONGENITAL DISEASE

10. TYPE OF GENE THERAPY SOMATIC CELL GENE THERAPY GERM LINE GENE THERAPY

11.  GENE THERAPY  ***REMOVE GENE FROM A SELECTED TISSUE  ***EXPOSITION TO THE GENE TRANFER VECTOR  ***SELECTION FOR TRANSGENIC USING MARKER  ***REINTRODUCTION THE CORRECTED IN TO THE PATIENT BODY

12. GENE THERAPY GENE THERAPY INJECTION THE VECTOR DIRECTLY IN TO THE TARGETED TISSUE INJECTION THE VECTOR DIRECTLY IN TO THE TARGETED TISSUE

13. BROAD METHODS ***REPLACE A NON FUNCTIONAL GENE ***HOMOLOGOUS RECOMBINATION ***SELECTIVE REVERSE MUTATION ***THE ALTERATION OF THE GENE REGULATION

14. To increase specificity and safety of gene therapy needs to: Gene delivery using viral and non-viral method. Targeting gene expression Condition targeted expression Disease targeted expression

15. Targeted delivery Molecular chemotherapy Delivery of suicide gene Gene directed enzyme Prodrug therapy Genetic immunopotentiation Mutation compensation

16. Viral and non-viral 1. methods Viral-mediated methods retrovirusHVJ Adeno virus Lipid-mediated methods Liposome The others Micro injection Microparticle bombardment

17. LIPOLEXES ANIONIC NEUTRAL CATIONIC

18. POLYLEXES CHITOSAN TRIMETHL CHITOSAN

19.  HYBRID METHODS  VIROSOMES  (LIPOSOME +INACTIVATED HIV)

20. DENDRIMERS

21. Non solid tumours: Herpes simplex virus thymidine kinase and cytosine deaminase Herpes simplex virus thymidine kinase and cytosine deaminase Modified envelope proteins Modified envelope proteins Targeted delivery of DNA via receptor Targeted delivery of DNA via receptor Solid tumours: Solid tumours: An aerobic bacteria An aerobic bacteria

29. Retroviral vectors with modified envelope proteins Mo-MuLV SU Binding of the virus to Its cell receptor TM Anchors the molecule to the viral membrane

31. SPECIFIC TARGETING STRATEGIES OF CANCER GENE THERAPY USING scFv RETROVIRAL VECTOR DISPLAYS BOTH ANTI CEA scFv AND Inos gene CHIMARIC RECEPTOR GENE WHICH ENCODED AN NATI CEA scFv AND THE ZETA CHAIN OF TCR/CD3

32. Delivery of suicide gene

33. Targeted delivery of DNA via receptor

34. REDIRECTING VIRUS TO A TISSUE SPECIFIC RECEPTOR REDIRECTING VIRUS TO A TISSUE SPECIFIC RECEPTOR USING TISSUE SPECIFIC LIGANDS OR mAb INCORPORATED ON TO THE SURFACE OF LIPOSOME TO DIRECT THEM TO TARGET CELLS USING TISSUE SPECIFIC LIGANDS OR mAb INCORPORATED ON TO THE SURFACE OF LIPOSOME TO DIRECT THEM TO TARGET CELLS

38. Targeted delivery and receptor overexpressing human cells EGF-R Folate-R C-kit-R

39. VDEPT Virally directed enzyme prodrug therapy

41. SYNTHETIC OLIGONUCLEOTIDES ANTISENSE siRNA DOUBLE STRANDED OLIGODEOXY NUCLEOTIDES

43. PEPTIDE NUCLEIC ACID SEQUENCE SPECIFIC RECOGNITION IN TARGETING GENE THERAPY TO CANCER

46. PNA IN CANCER GENE THERAPY ***INHIBITION OF TELOMERASE ACTIVITY ***ANTISENSE AND ANTIGENE ACTIVITY OF PNA TO HUMAN B-CELL LYNPHOMA ***INHIBITION OF HIV REVERSE TRANSCRIPTASE ***PNA AS A GENETIC ANTIBIOTICS ***DIHYDROTESTOSTRONE LINKED TO PNA AS A VECTOR FOR TARGETING C-MYC DNA TO PROSTATIC CANCER CELLS

47. EFFECTIVE METHODS TO INDUCE UPTAKE OF PNA IN TO CELLS ***ELECTROPORATION ***ELECTROPORATION ***CATIONIC LIPIDS ***CATIONIC LIPIDS ***ADENO VIRUS-POLYLYSINE COMPLEXES ***ADENO VIRUS-POLYLYSINE COMPLEXES ***STEREPTOLYSINE ***STEREPTOLYSINE

48. TRANSCRIPTIONAL CONTROL OF EXPRESSION

51. T0MOUR SPECIFIC PROMOTERS 1)PROMOTERS THAT SPECIFIC ONLY IN MALIGNANT PROCESS 1)PROMOTERS THAT SPECIFIC ONLY IN MALIGNANT PROCESS 2)PROMOTERS THAT ONCOFOETAL RELATED WITH TISSUE SPECIFICITY. 2)PROMOTERS THAT ONCOFOETAL RELATED WITH TISSUE SPECIFICITY. 3)TOMOUR MICROENVIROMENT-RELATED PROMOTERS 3)TOMOUR MICROENVIROMENT-RELATED PROMOTERS 4)TOMOUR VASCULATURE-RELATED PROMOTERS. 4)TOMOUR VASCULATURE-RELATED PROMOTERS.

55. Tissue targeted expression

56. Alpha feto protein to target hepatocellular carcinoma

57. AFP has been found to be abnormally activated in hepatocellular carcinoma. Transgenic mice carrying the SV40 large T-antigen developed carcinoma crossed with mice transgenic for HSV thymidine kinase. HSVtk activate GCV under the AFP promoter/enhancer control.

58. Tissue targeted expressionTissue targeted expression Albumin enhancer to target liver cancerAlbumin enhancer to target liver cancer

59. Retroviral vector containing the liver-specific expression system. The albumin enhancer element and promoter to target expression of B-gal to hepatoma cells. Expression could only be detected in dividing hepatocytes.

60. Condition targeted expression

61. Tissue-type plasminogen activator regulated by radiation.  t-pA induces over 50 fold after irradiation with x-rays.  The t-pA protease have a function equivalent to the sos repair system.

62. GRP encoding gene was used to control expression of a marker gene in a murine fibrosarcoma model. In vitro glucose deprivation of transduced fibrosarcoma cells showed an 8-fold induction over non-stressed cells.

63. Hypoxia regulated gene expression (in almost solid tumours)

64. Hypoxic conditions can modulate the expression of a number genes: EEEEncoding growth factors. TTTTranscription factors. GGGGlycolytic and DNA repair enzymes.

65. Hypoxic expression is controlled by the binding of transcription factor(HIF-1) to a DNA sequence which can be enhancer or response element.(HRE) Hypoxic expression is controlled by the binding of transcription factor(HIF-1) to a DNA sequence which can be enhancer or response element.(HRE)

66. The use of hypoxia enhancer elements increase the levels of the marker gene CD2. Hypoxic induction in vivo of the CD2 was confirmed by CD2 staining with the comet assay.

67. In comet assay,tumour cells are treated with radiation and a bioreductive drug(which indices DNA crosslinks only in hypoxic cells) Electrophoresis of single cells isolated from the treated tumours can differentiate the two populations.

68. The hypoxic cells,according to the comet assay,stained positive for CD2,whereas the aerobic ones did not. This result demonstrate the selectivity of the system and its potential for tumour specific targeting of gene expression.

69. INDUCTION OF APOPTOSIS IN HYPOXIC CONDITION P53BAXhREC2Caspase-8

70. Genetically modified tomour vaccines in gene therapy

71. Cytokine genes

72. Co-stimulatory molecules

74. DNA VACCINE

75. Conclusion

76. Gene therapy has the potential to have few side effects and lower systemic toxicity than current therapies. Also choosing only one criteria for selectivity such as targeting delivery to cells or tissue specific expression,is not sufficient.

77.  Only by combining the most successful strategies in cancer gene therapy approaches,will be a successful clinical treatment emerge.