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  • Gene Therapy: Young Jik Kwon, PH.D

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    This document discusses various approaches to cancer gene therapy, including enhancing the immune system response, inactivating oncogenes, and restoring tumor suppressor genes like p53. It also covers molecular chemotherapy using genes like HSV/TK, and compares somatic cell gene therapy with stem cell gene therapy. Key differences noted are that somatic therapy is temporary while stem cell therapy can result in permanent changes, and stem cell therapy requires viral vectors. RNA interference techniques like siRNA and challenges associated with them are also summarized.

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    This document discusses various approaches to cancer gene therapy, including enhancing the immune system response, inactivating oncogenes, and restoring tumor suppressor genes like p53. It also covers molecular chemotherapy using genes like HSV/TK, and compares somatic cell gene therapy with stem cell gene therapy. Key differences noted are that somatic therapy is temporary while stem cell therapy can result in permanent changes, and stem cell therapy requires viral vectors. RNA interference techniques like siRNA and challenges associated with them are also summarized.

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    17 views17 pages

    Gene Therapy: Young Jik Kwon, PH.D

    Uploaded by

    fakemotha247
    This document discusses various approaches to cancer gene therapy, including enhancing the immune system response, inactivating oncogenes, and restoring tumor suppressor genes like p53. It also covers molecular chemotherapy using genes like HSV/TK, and compares somatic cell gene therapy with stem cell gene therapy. Key differences noted are that somatic therapy is temporary while stem cell therapy can result in permanent changes, and stem cell therapy requires viral vectors. RNA interference techniques like siRNA and challenges associated with them are also summarized.

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    of 17

    Gene Therapy

    October 20, 2011

    Young Jik Kwon, Ph.D.


    132 Sprague Hall (949) 824-8714 kwonyj@uci.edu

    1
    PhrmSci M174/CBEMS 195 Biopharmaceutics, Fall 2011

    Cancer Gene Therapy


    Immune activation
    Enhancement of the response of the immune system to cancers. e.g., Introduction of IL-2 and TNF- to tumor infiltrating lymphocytes

    Oncogene inactivation
    Employed for dominantly inherited monogenic diseases. e.g., Oligodeoxynucleotides target the promoter regions of oncogenes. Antisene techniques prevent transport & translation of the oncogene mRNA.

    Restoration of tumor suppressor


    p53: cellular apoptosis & arrest tumor growth

    Molecular chemotherapy
    Introduction of a gene coding for a toxic product. e.g., Herpes simplex virus thymidine kinase (HSV/TK)
    2
    PhrmSci M174/CBEMS 195 Biopharmaceutics, Fall 2011

    The Immune System: A Popular Target of Gene Therapy

    Nat. Immunol. 5, 971 - 974 (2004)


    3
    PhrmSci M174/CBEMS 195 Biopharmaceutics, Fall 2011

    Melanoma Gene Therapy


    Patient 4, liver

    Patient 14, LN

    Morgan et al., Science 2006


    PhrmSci M174/CBEMS 195 Biopharmaceutics, Fall 2011

    Apoptosis Pathways

    5
    PhrmSci M174/CBEMS 195 Biopharmaceutics, Fall 2011

    p53-mediated Cancer Gene Therapy

    Mice aerosol delivery of PEI-p53 plasmid DNA polyplexes

    Needle-free delivery

    Too high and long expression of p53: collapsing the lung

    For cancer, nonviral vector and adnoviral vectors have advantages


    Densmore et al., Cancer Gene Ther 8, 619 - 671 (2001) PhrmSci M174/CBEMS 195 Biopharmaceutics, Fall 2011

    Somatic vs Stem Cell Gene Therapy


    Somatic cell gene therapy

    Somatic vs Stem Cell Gene Therapy


    Stem cell gene therapy

    Target

    Acquired disorders

    Inherited disorders

    Modification

    Temporary

    Permanent

    Vectors

    Viral and nonviral

    Viral vectors only (RV, LV)

    Routes

    Ex vivo and in vivo

    Ex vivo

    Mutagenesis

    Low

    High

    7
    PhrmSci M174/CBEMS 195 Biopharmaceutics, Fall 2011

    Stem Cell Gene Therapy of SCID


    DNA RNA

    Cavazzana-Calvo et al., Science, 2000

    8
    PhrmSci M174/CBEMS 195 Biopharmaceutics, Fall 2011

    Generation of Stem Cells


    Four transcription factorsOct4 (also called Oct3/4 or Pou5f1), Sox2, c-Myc and Klf4, delivered by MoMLV to murine fibroblasts.

    Wernig et al., Nature, 2007

    9
    PhrmSci M174/CBEMS 195 Biopharmaceutics, Fall 2011

    Antisense Drugs: Oligonucleotides


    Gene-selective reagents and drugs

    Rational design of reagents and drugs based on Watson-Crick hybridization

    Broadened potential targets and pathways of noncoding RNAs, RNA structure, function, and the metabolism

    10
    PhrmSci M174/CBEMS 195 Biopharmaceutics, Fall 2011

    Challenge of Antisense Drugs:


    New pharmacology: Medicinal chemistry

    Crooke et al., Antisense drug technology, 2008

    11
    PhrmSci M174/CBEMS 195 Biopharmaceutics, Fall 2011

    Mechanisms
    Occupancy-only-mediated mechanisms Modulation of splicing

    Translation arrest

    Disruption of necessary RNA structure

    Occupancy-activated destabilization Inhibition of 5 Capping Inhibition of 3-polyadenylation

    Other mechanisms: Rnase H, siRNA


    PhrmSci M174/CBEMS 195 Biopharmaceutics, Fall 2011

    12

    Ribonuclease (RNase)
    The most robust mechanism identified and characterized to date

    Cleaves only in RNA-DNA duplexes

    Two cloned human RNase H1 and H2

    13
    PhrmSci M174/CBEMS 195 Biopharmaceutics, Fall 2011

    RNA Interference
    Craig C. Mello and Andrew Fire,1998

    Nobel prize in 2006

    Nature 418, 244 (2002)

    Nature Chemical Biology 12, 711 (2006)

    PhrmSci M174/CBEMS 195 Biopharmaceutics, Fall 2011

    Small Interfering RNA (siRNA)


    Double-stranded RNase

    RNA-induced silencing complex (RISC)-mediated pathways

    Challenges of duplex RNA drugs Physical chemical properties Manufacturing The sense strand Immune stimulation

    15
    PhrmSci M174/CBEMS 195 Biopharmaceutics, Fall 2011

    Comparison of RNase H and siRNA

    16
    PhrmSci M174/CBEMS 195 Biopharmaceutics, Fall 2011

    17
    PhrmSci M174/CBEMS 195 Biopharmaceutics, Fall 2011

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