Microsoft PowerPoint - 9 Gene therapy.pptx

Size: px

Start display at page:

Download "Microsoft PowerPoint - 9 Gene therapy.pptx"

帐锥郎
5 years ago
Views:

1 Pharmaceutical Biotechnology Gene Therapy By Yuqiong Xia

2 Keywords Gene therapy, in vivo, ex vivo Vector, vascular disease, monogenetic disease, infectious disease, AIDS, HIV Gene expression cassette, gene gun, cationic polymers, liposomes 2

3 Outline Introduction to gene therapy In vivo and ex vivo gene therapy Gene therapy in clinics Vectors for gene transfer 3

4 Introduction to gene therapy The use of nucleic acids as therapeutic medicinal compounds Compensate for abnormal gene expression 4

5 The first gene therapy Ashanti DeSilva was born with a lack of the enzyme adenosine deaminase (ADA). The root cause is a single defective gene. Patients with ADA deficiency suffer from severe immunodeficiency. 5

6 6

7 The first gene therapy Dr. Anderson s team drew blood from their patient, and replaced the defective gene with a functional variant. The therapy partially restores Ashanti s immune system. It temporarily stimulates production of the missing enzyme, but does not generate new cells with functional genes. Ashanti continues to receive injections of corrected T- cells every two months. She is able to lead a normal life. 7

8 Outline Introduction to gene therapy In vivo and ex vivo gene therapy Gene therapy in clinics Vectors for gene transfer 8

9 In vivo and ex vivo gene therapy 9

10 10

11 In vivo vs ex vivo In vivo Direct injection of vector/dna complexes into bloodstream Intratumoral, subcutaneous, intramuscular injection Ex vivo Isolation and culture of cellular targets Direct application of the vector for efficient gene expression 11

12 Outline Introduction to gene therapy In vivo and ex vivo gene therapy Gene therapy in clinics Vectors for gene transfer 12

13 13

14 Disease target 生物制药工程夏玉琼西安电子科技大学 14

15 Cancer gene therapy Correction of genetic mutations Immunotherapy Use of oncolytic viruses 15

16 Correction of genetic mutations Over-expression of tumor suppressor genes such as p53, MDA-7, ARF p53 is one of the most studied candidates 16

17 The first gene therapy product 生物制药工程夏玉琼西安电子科技大学 17

18 18 Immunotherapy to treat cancer Limitations Limited expression and low avidity of tumorassociated antigen-specific cytotoxic T-cells Inherent ability of tumor cells to evade immune detection Methods Gene transfer to cancer tissue Gene transfer to cancer cells Gene transfer to T-cells

19 Immunotherapy to treat cancer 生物制药工程夏玉琼西安电子科技大学 19

20 Use of oncolytic viruses Oncolytic viruses induce cell death of tumor cells Through replication, expression of cytotoxic proteins and cell lysis in tumor cells Remaining safe to normal cells. 20

21 Oncolytic viruses Vaccinia( 牛痘 ) HSV I( 疱疹 ) 呼肠孤病毒新城病毒小儿麻痹病毒腺病毒

22 Gene therapy for vascular disease Reduce blood pressure Overexpression of genes involved in vasodilation( 血管舒张 ) Inhibition of molecules involved in vasoconstriction ( 血管收缩 ) 22

23 Gene therapy for coronary and peripheral ischemia Over-expression of pro-angiogenic factors ( 促进血管生成因子 ) Over-expression of endothelial and inducible NOS (cause vasodilation) Over-expression of genes that can reduce cholesterol 23

24 Monogenetic inherited disorders Of the 25,000 genes that comprise human genome, mutations in over 1,800 have been identified as the cause of a variety of hereditary orders Replace a gene with its functioning counterpart in stem cells 24

25 Monogenetic inherited disorders Limitations Lack of suitable gene delivery technologies Unfavorable interactions between the host and gene transfer vector Complex biology and pathology of monogentic disease and target organs Lack of relevant measures to assess the clinical efficacy of gene transfer 25

26 Gene therapy for infectious disease AIDS Methods Over expression of proteins that interfere with HIV replication RNA-based strategies 26

27 Outline Introduction to gene therapy In vivo and ex vivo gene therapy Gene therapy in clinics Vectors for gene transfer 27

28 Why gene transfer need a vector? 28

29 The transferred gene Gene expression cassette for gene therapy 29

30 Vectors for gene transfer Virus vectors Non-viral vectors Physical methods Chemical methods 30

31 Structure of Retrovirus 生物制药工程夏玉琼西安电子科技大学 31

32 Simple and complex retrovirus genome 32

33 The retrovirus replication cycle 生物制药工程夏玉琼西安电子科技大学 33

34 Advantages of retroviral vector Accommodate transgene cassettes of 8 kb Integrate into the host genome Pseudotyping Surface glycoproteins are replaced with those from unrelated virus 34

35 Disadvantages of retroviral vector Cannot transduce non-dividing cells May disrupt normal cellular processes The virus titer is very low Virus particles are generally unstable Rapidly removed from the systemic circulation 35

36 Adenoviral vectors 生物制药工程夏玉琼西安电子科技大学 36

37 The adenovirus infection cycle 生物制药工程夏玉琼西安电子科技大学 37

38 Advantages and disadvantages Advantages Biology of the virus is well understood Transgene expression is rapid and robust Physically stable Can infect dividing and non-dividing cells Disadvantages Elicit a strong immune response 38

39 Physical methods: gene gun 39

40 Physical methods: electroporation 40

41 Physical methods: sonoporation 41

Physical methods: laser irradiation 42 http://www.

42 Physical methods: laser irradiation 42

43 Physical methods: magnetofection 43

44 Chemical methods: cationic polymers 44

45 Chemical methods: liposomes 生物制药工程夏玉琼西安电子科技大学 45

46 Non-viral vectors for gene therapy Advantages Unlimited cloning capacity Reduced capacity for insertional mutagenesis Easy to manipulate Inexpensive Disadvantages Low transduction efficiency Poor delivery Limited capacity 46

47 Summary In vivo and ex vivo gene therapy: the differences Gene therapy in cancer: three methods Vector for gene therapy Gene expression cassette Non-viral vectors: gene gun, cationic polymers, liposomes 47

20110420

20110420 愛滋病與結核病防治台北榮民總醫院感染科國立陽明大學醫學系王永衛醫師它是什麼? 愛滋病毒 (HIV) 後天免疫缺乏病毒愛滋病 (AIDS) 後天免疫缺乏症候群愛滋病 1980 年迄今 Million 50 45 40 35 30 25 20 15 1 First cases of