01. Gene therapy Boulikas.pdf - Gene therapy & Molecular Biology

More documents

Recommendations

Info

natriuretic peptide (ANP) gene IL-2 prostate cancer liposom es mouse leptin cDNA rat leptin cDNA/CMV VEGF 165 cGMP phosphodiest erase-β (PDE-β) gene Boulikas: An overview on gene therapy natriuresis, diuresis, and hypotension blood pressure in young hypertensive rats (4 weeks old); the effect continued for 7 weeks. Direct transfer of the IL-2 gene under control of the CMV promoter with or without the AAV inverted terminal repeats obesity Adeno ob/ob mouse (which is genetically deficient in leptin and exhibits both an obese and a mild non-insulin-dependent diabetic phenotype) obesity Adeno Wistar rats infused with 8 ng/ml adeno/leptin cDNA for 28 days cancer (vascularizatio n) retinal degeneration calcium phospha te XXIX. Gene therapy of HIV Expression of VEGF 165 in rat C6 glioma cells and subcutaneous injection of the transduced cells in athymic mice. AAV To treat retinal degeneration due to recessive mutation in the endogenous gene A. Mechanism of HIV-1 entry into T cells and macrophages Targets of human immunodeficiency virus (HIV) are helper T cells and macrophages; macrophage-tropic HIV-1 isolates represent the most prevalent phenotype isolated from individuals shortly after seroconversion during the asymptomatic period of the disease; tropism is determined by specific sequences in the third variable loop (V3 domain) of gp120 coat protein of HIV-1. The CD4 receptor on both macrophages and T cells is the primary receptor mediating HIV-1 entry into the cell; however, HIV-1 was unable to infect CD4 + T cells of mice engineered to express human CD4 (reviewed by D’Souza and Harden, 1996). Thus, a second chemokine receptor was thought to be necessary for HIV entry into immortalized T cell lines. The second receptor for entry of HIV-1 into T cells and macrophages is CCR-5, a β-chemokine receptor; CCR-5 is a seven transmembrane-domain glycoprotein of the chemokine superfamily of receptors related to the receptor of IL-8 (G protein-coupled proteins that transduce signals from the cell surface to the interior of cells). The β-chemokines RANTES, MIP-1α, and MIP-1β inhibited replication of M-tropic isolates of HIV-1 and were found to be major HIV-suppressive factors produced by CD8 + T cells. The V3 loop of gp120 determines interaction with the chemokine receptor. CD4, in addition to providing a 94 Plasmid DNA containing the AAV inverted terminal repeats showed 3- 10 fold higher levels of gene transfer and IL-2 expression compared with constructs lacking the AAV sequences. Dramatic reductions in both food intake and body weight, as well as in normalization of serum insulin levels and glucose tolerance Animals became hyperleptinemic; 30-50% reduction in food intake; gained only 22 g over the experimental period versus 115-132 gained by control animals Tumors from cells expressing VEGF grew slower than tumors developed from nontransduced C6 cells, were highly vascularized, and contained varying degrees of necrosis and eosinophilic infiltrate Intraocular injection of AAV-PDE-β cDNA increased retinal expression of immunoreactive PDE protein; treated eyes showed increased numbers of photoreceptors and a two-fold increase in sensitivity to light Vieweg et al, 1995 Muzzin et al, 1996 Chen et al, 1996 Saleh, 1996 Jomary et al, 1997 docking surface for the gp120 glycoprotein of HIV-1 promotes exposure of the V3 domain on gp120 that can interact with the chemokine receptor CCR-5 (Scarlatti et al, 1997; reviewed by D’Souza and Harden, 1996). A small number of individuals (1% in populations of European descent but much lower in non-Caucasian populations) remain uninfected despite multiple high risk exposures; such individuals have a homozygous defect in the CCR-5 receptor gene which consists of a 32-bp deletion in the region encoding the second extracellular loop of the receptor; the defective protein is not detected at the cell surface; this defect prevents the proper interaction and entry of HIV-1 into their T cells and macrophages (Liu et al, 1996). This defect precludes infection from HIV-1 from all routes. The disease progresses much slower in heterozygotes for the 32-bp deletion (18% in populations of European descent) who are not protected from HIV-1 infections. This finding offers the hope of reconstituting the immune system of HIV-infected individuals with CD34 + stem cells from fetal cord blood or stem cells and lymphoid tissue from individuals who carry the homozygous deletion in the CCR-5 gene, an approach to deal with the immune rejection problems in patients with heart and kidney transplants. The identification of chemokine receptors and their role in HIV-1 infections has closed a major gap in AIDS research; transgenic animals can now be produced expressing both human CD4 and chemokine receptors to evaluate the efficacy of AIDS therapeutics and testing vaccines; new prophylactic or therapeutic vaccines can be
designed by immunization with portions or the entirety of CCR-5 and/or gp120 to generate appropriate antibodies (D’Souza and Harden, 1996). Inactivation of the CCR-5 co-receptor to mimic the natural resistance of the CCR-5defective individuals, in cultured lymphocytes, rendered them viable and resistant to macrophage-tropic HIV-1 infection (Yang et at, 1997). B. Therapeutic strategies against HIV A number of therapy strategies emerged soon after identification of HIV as the etiologic agent of AIDS. Virtually every stage in the viral life cycle and every viral gene product is a potential target. Albeit major efforts for the combat of HIV have focused on the development of antiviral drugs and preventive vaccines, a number of studies have been aimed at eliminating HIV with gene therapy. The intracellular immunization approach (Baltimore, 1988) has prompted the advent of molecular tactics for inhibiting replication and infection of HIV (Trono et al, 1989; Malim et al, 1989). Four main targets have been defined in HIV therapeutics: (i) viral RNAs using ribozymes and antisense RNAs; (ii) viral proteins using RNA decoys, trans-dominant viral proteins, intracellular single-chain antibodies, and soluble CD4; (iii) infected cells aimed at eliminating those with transfer and expression of suicide genes; and (iv) the immune system by in vivo immunization (see Corbeau et al, 1996). Such gene therapeutic approaches can be combined with potent antiretroviral drugs especially the potent reverse transcriptase and protease inhibitors (Junker et al, 1997). The elucidation of the chemokine receptor mechanisms for entry of HIV-1 into T cells and macrophages provides new tactics for intervention at the level of interaction of gp120 with CCR-5. Inactivation of the CCR-5 gene might be achieved (i) with triplex oligonucleotide technologies once critical transcription factor binding sites in the regulatory region of the gene have been determined; (ii) with saturation of the blood of infected individuals with ligands, selected from peptide libraries, that block the extracellular domains of the CCR-5 receptor and preclude interaction with gp120; (iii) with antagonists of RANTES (see above) that lack chemotactic activity but can block HIV infections (Arenzana-Seisdedos et al, 1996). Specificity for the ablation of HIV Tat-expressing cells has been achieved through the use of the promoter element from the long terminal repeat (LTR) of HIV (Venkatesh et al, 1990; Caruso et al, 1992). C. Gene therapy against HIV in cell culture Strategies for HIV gene therapy include the inactivation of the CCR-5 coreceptor which is accomplished by targeting a modified CC-chemokine to the endoplasmic reticulum to block the surface expression Gene Therapy and Molecular Biology Vol 1, page 95 95 of newly synthesized CCR-5 (Yang et al, 1997). A different gene therapy strategy proposed is targeting Tat, an early regulatory protein that is critical for viral gene expression and replication and which transactivates the LTR of HIV-1 via its binding to the transactivation response element (TAR); Tat also superactivates the HIV- 1 promoter via activation of NF-κB in a pathway involving protein kinase C and TNF-α; combinations of the NF-κB inhibitors, pentoxifylline and Go-6976, with a stably expressed anti-Tat single-chain intracellular antibody suppressed HIV-1 replication and LTR-driven gene expression (Mhashilkar et al, 1997). Production of recombinant retrovirus containing the HSV-tk gene coupled to the HIV-2 promoter and Tat responsive region (TAR) has been used on human and mouse cells in culture for the specific elimination of HIV Tat-expressing cells; since the HIV-2 promoter can sustain a considerable level of basal expression in the absence of its activator, Tat, a number of modifications were made to the HIV-2 promoter in order to minimize toxicity to non-infected cells. Retroviruses export unspliced, intron-containing RNA to the cytoplasm of infected cells despite the fact that intron-containing cellular RNAs cannot be exported; this export pathway is a critical step in the HIV-1 life-cycle. In HIV-1 this is accomplished through an interaction between the viral regulatory Rev protein and the Rev response element (RRE) RNA. In the absence of Rev, these intron-containing HIV-1 RNAs are retained in the nucleus. The nuclear export sequence (NES) LQLPPLERLTL has been identified on Rev that is responsible for its export to the cytoplasm (see Boulikas, 1998, this volume for more details). Targeting of Rev has provided a framework for novel interventions to reduce virus production in the infected host. Because disruption of either Rev or the RRE will completely inhibit HIV-1 replication, an anti-HIV-1 intracellular immunization strategy was developed based on RRE region-specific hammerhead ribozymes and on the intracellular expression of an anti-HIV-1 Rev single chain variable fragment (Sfv), which specifically targets the Rev activation domain. This combination resulted in a potent inhibition of HIV-1 replication in cell culture that holds promise as a future therapeutic regimen (Duan et al, 1997). To disable Rev function, primary T cells or macrophages were transduced with a recombinant AAV carrying an anti-Rev singlechain immunoglobulin (SFv) gene or an RRE decoy gene or with combinations of the two genes to disrupt the interaction between Rev and the RRE; when the transfected cells were then challenged by either clinical or laboratory HIV-1 isolates, this genetic antiviral strategy effectively inhibited infection (Inouye et al, 1997). A synergic effect of anti-Tat and anti-Rev molecules was found when the RRE sequence was cloned 3' to a tat transdominant negative mutant (tat22/37) gene; for this strategy Jurkat cells were transduced with the recombinant
Page 1 and 2:
Gene Ther Mol Biol Vol 1, 1-172. Ma
Page 3 and 4:
I. Introduction Monumental progress
Page 5 and 6:
The use of retroviral vectors in hu
Page 7 and 8:
displaying the cleavable EGF domain
Page 9 and 10:
molecules in the nucleus (Lowe and
Page 11 and 12:
sensitive mutations which allow pro
Page 13 and 14:
processed as described in panel A s
Page 15 and 16:
On the contrary, the payload capaci
Page 17 and 18:
in K562 human erythroleukemia cells
Page 19 and 20:
defective HSV virus (DeLuca and Sch
Page 21 and 22:
complex into the cytosol from the e
Page 23 and 24:
Gene Therapy and Molecular Biology
Page 25 and 26:
delivered by Sendai virus-liposome
Page 27 and 28:
plasmids with the cell surface, tra
Page 29 and 30:
infected by adenovirus alone; infec
Page 31 and 32:
Page 33 and 34:
B. Transcription factor binding sit
Page 35 and 36:
A closely related family of ubiquit
Page 37 and 38:
cells. I-CreI is a member of this c
Page 39 and 40:
Page 41 and 42:
C. Considerations of chromatin stru
Page 43 and 44: Gene Therapy and Molecular Biology
Page 45 and 46: A. The molecular basis of cancer im
Page 47 and 48: fragments, or heat shock proteins c
Page 49 and 50: ecombinant vaccinia virus expressin
Page 51 and 52: HIV-1 envelope DNA construct develo
Page 53 and 54: inding sites A GYYY oriented in opp
Page 57 and 58: Prives, 1994). Whereas the wild-typ
Page 59 and 60: mutant p53 (mutations on p53 are wi
Page 61 and 62: master regulator of the cell cycle
Page 63 and 64: Work from several groups has shown
Page 65 and 66: chromatin condensation, drop in pH,
Page 67 and 68: Figure 23. A pathway leading to the
Page 69 and 70: Excessive necrotic death in cells o
Page 71 and 72: implantation, and similar processes
Page 73 and 74: normal cells in the surroundings. T
Page 79 and 80: A bicistronic retroviral vector (Ha
Page 81 and 82: C. Antisense ras gene transfer for
Page 83 and 84: equal to 6.0 showed S1 hyperreactiv
Page 85 and 86: protein (e.g. Smith et al, 1995; Fo
Page 87 and 88: transduced primary mouse fibroblast
Page 89 and 90: IL-1 - receptor antagonist protein
Page 91 and 92: p53 lung cancer retroviru s MHC cla
Page 93: Gene Therapy and Molecular Biology
Page 97 and 98: Isolation of an RNA aptamer that ca
Page 99 and 100: induction of human apoE in neonate
Page 101 and 102: atrium in heart, endothelial cells
Page 103 and 104: which eliminated tumors in small an
Page 105 and 106: C. Transfer of other genes against
Page 109 and 110: significant reduction in neointima
Page 111 and 112: (Prehn et al, 1996); this implies a
Page 113 and 114: B. Approaches to gene therapy of RA
Page 115 and 116: of human immunoglobulin and antigen
Page 117 and 118: A. Etiology and mechanisms of destr
Page 121 and 122: However, CsA also inhibits the acti
Page 123 and 124: The peptide kinin, after binding to
Page 125 and 126: intake when administered to adrenal
Page 127 and 128: Introgen Therapeutics (Austin and H
Page 129 and 130: When a subfragment of only 513 bp o
Page 131 and 132: Armentano D, Zabner J, Sacks C, Soo
Page 133 and 134: Brady HJM, Miles CG, Pennington DJ,
Page 135 and 136: Chen J, Stickles RJ, Daichendt KA (
Page 137 and 138: Day ML, Wu S, Basler JW (1993) Pros
Page 139 and 140: Farmer G, Bargonetti J, Zhu H, Frie
Page 141 and 142: Giovannangeli C, Perrouault L, Escu
Page 143 and 144: the neoplastic phenotype by replace
Page 145 and 146:
integrate in a site-specific fashio
Page 147 and 148:
Li R, Waga S, Hannon GJ, Beach D, S
Page 149 and 150:
adiation-induced apoptosis in the g
Page 151 and 152:
Nakaya T, Iwai S, Fujinaga K, Sato
Page 153 and 154:
Polyak K, Xia Y, Zweier JL, Kinzler
Page 155 and 156:
macrophages from simian immunodefic
Page 157 and 158:
intimal hyperplasia in a rat caroti
Page 159 and 160:
Trono D, Feinberg MB, Baltimore D (
Page 161 and 162:
Wattiaux R, Jadot M, Warnier-Pirott
Page 163 and 164:
similar to mammalian interleukin-1
Page 165 and 166:
Page 167 and 168:
24. Gene Marking /Infectious Diseas
Page 169 and 170:
Drug 50. Gene Therapy /Phase I /Can
Page 171 and 172:
76. Gene Therapy /Phase I /Cancer /
Page 173 and 174:
99. Gene Therapy /Phase II /Cancer
Page 175 and 176:
120. Gene Therapy /Phase I /Monogen
Page 177 and 178:
Page 179 and 180:
cancer not specified) /Immunotherap
show all

01. Gene therapy Boulikas.pdf - Gene therapy & Molecular Biology

Create successful ePaper yourself

Delete template?

Save as template?