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

More documents

Recommendations

Info

The replication and expression of hepatitis B virus (HBV) could be inhibited through antisense gene transfer and this could become a new method for clinical gene therapy against HBV; infection of the human hepatoblastoma cell line 2.2.15, which expresses HBV surface antigen and releases HBV particles, with retroviral vectors carrying an antisense preS/S or preC/C genes of HBV inhibited expression of the surface antigen (Ji and St 1997). Phosphorothioate antisense oligos directed against cmyc and p53 in different cell lines (CAOV-3, SKOV-3, and BG-1) were shown to have both antiproliferative and stimulatory activity, as single agents and in combination; it was concluded that further in vitro studies are needed before considering clinical trials with these agents in gynecologic cancers (Janicek et al, 1995). Transfection of antisense cDNA constructs encompassing different regions of the c-erbB-2 gene in the lung carcinoma cell line Calu3, which overexpresses the cerbB-2 oncogene, reduced significantly anchorageindependent growth and tumor size in nude mice (Casalini et al, 1997). Antisense oligonucleotides against PCNA and cdc2 kinase transferred into injured arterial walls by proteinliposomes greatly reduced mRNA levels for those genes and inhibited neointima formation of the injured artery for 8 weeks; double-stranded oligonucleotides containing the consensus sequence for E2F binding sites also inhibited the growth of smooth muscle cells and prevented neointima formation (Kaneda et al, 1997). Antisense oligonucleotides to angiotensinogen1-receptor mRNA and to angiotensinogen mRNA reduced blood pressure (Tomita et al, 1995; Phillips, 1997; Phillips et al, 1997). XXVI. Triplex gene therapy Boulikas: An overview on gene therapy 82 Figure 29. Photographs of nude mice treated with antisense cyclin G vector (panel A) have smaller tumors that animal treated with a control vector (panel B). Panel C: the relative tumor size (% of day 0 tumor size divided by 100) is plotted, on the vertical axis, as a function of time (days), plotted on the horizontal axis. From Chen DS, Zhu NL, Hung G, Skotzko MJ, Hinton DR, Tolo V, Hall FL, Anderson WF, Gordon EM (1997) Retroviral vector-mediated transfer of an antisense cyclin G1 construct inhibits osteosarcoma tumor growth in nude mice. Hum Gene Ther 8, 1667-1674. Reproduced with kind permission from the authors and Mary Ann Liebert, Inc. A. Molecular mechanisms for triplex formation Natural purine . pyrimidine sequences in regulatory regions of genes in eukaryotic cells with a mirror symmetry can form triple-helical structures; in addition, purine-rich segments in DNA unable to form triple helices on their own can be targeted by DNA or RNA oligonucleotides able to form triplex structures with their target DNA and these unusual structures can inhibit transcription factor binding, transcription initiation, and nuclear enzymatic activities. Understanding the advantages, limitations and pitfalls for using oligonucleotides as gene bullets, development of strategies for boosting their therapeutic efficiency, their covalent linkage to DNA damaging molecules to hit a specific genomic DNA sequence, and improvements to the methods for their delivery to cells could make reality their use as tools of micro-targeting specific genomic sites and as pharmacogenomic drugs. Formation of triple helical DNA was found to take place on AT- and GC-rich stretches. A pyrimidine third strand oligonucleotide, studied by NMR and other approaches, interacts with purine residues in the major groove of the target duplex in a parallel orientation (Moser and Dervan, 1987; Rajagapol and Feigon, 1989; de los Santos et al, 1989) whereas a purine oligonucleotide binds in an antiparallel orientation relative to the purine strand in the duplex (Cooney et al, 1988; Kohwi and Kowhi- Shigematsu, 1988; Beal and Dervan, 1991). In this case G can recognize GC pairs and A or T can recognize AT pairs. Specificity is provided from T . AT and C + GC base triplets where the bases of the third polypyrimidine strand establish Hoogsteen base pairing with the purine strand of the duplex (Hoogsteen, 1959; Rajagopal and Feigor, 1989). The H form is the structural basis for S1-nuclease hypersensitivity (Mirkin et al, 1987). A restriction fragment from a human U1 gene containing the sequence d(C-T) 18.d(A-G) 18 under supercoiling and pH less than or
equal to 6.0 showed S1 hyperreactivity in the center and at one end of the (C-T) n tract, and continuously from the center to the same end of the (A-G)n tract providing strong support for a triple-helical model (Johnston, 1988). Homopyrimidine oligodeoxyribonucleotides with EDTA-Fe attached at a single position bound the corresponding homopyrimidine-homopurine tracts within large double-stranded DNA by triple helix formation and cleaved at that site (Moser and Dervan, 1987). Studies from the group of Claude Hélène have similarly focused on the development of artificial scissor oligonucleotides based on triplex technology (Praseuth et al, 1988; Perrouault et al, 1990). However, the feasibility of employing this exciting in vitro technology to animal studies has not yet been demonstrated. Intramolecular or intermolecular triple helices could be recognized by specific proteins that stabilize triplex structures and might play a role in gene regulation; a protein from HeLa cell nuclear extracts was identified that binds to a 55 nucleotide-long DNA oligomer that could fold on itself to form an intramolecular triple helix of the Py Pu x Py motif (Guieysse et al, 1997). Triplex-forming oligophosphoramidates containing thymines and cytosines or 5-methyl cytosines (5' T 4CT 4C 6T 3') bind strongly to a 16 base pair oligopurine . oligopyrimidine sequence of HIV proviral DNA even at neutral pH and are remarkably stable compared to oligonucleotides with natural phosphodiester linkages. The phosphoramidate oligomers induced an efficient arrest of both bacteriophage and eukaryotic transcriptional machineries (SP6, T7 or Pol II) under conditions where the phosphodiester oligos had no inhibitory effect and blocked the RNA polymerases at the triplex site (Giovannangeli et al, 1996). Oligonucleotide-directed triplex formation has been shown to inhibit binding of transcription factors to their cognate DNA sequences. A 21 bp homopurine element insert flanking a single Sp1 site in the adenovirus E4 promoter was used to study the effect of oligo targeting on transcriptional efficiency in vitro; assembly of the triple helical complex repressed basal transcription by rendering the triplex target inflexible and by blocking assembly of the promoter into initiation complexes; Sp1 was unable to cause derepression (Maher et al, 1992). Thus DNA triplexes can inhibit transcription initiation not only when directed to a TF binding site occluding its binding but also to a flanking region by other possible repression mechanisms including stiffening of the double helix (Maher et al, 1992). B. Triplex targeting of IGF-I Oligonucleotide-directed triple helix formation targeted toward IGF-I to inhibit its expression was studied following stable transfection of C6 rat glioblastoma cells Gene Therapy and Molecular Biology Vol 1, page 83 83 with a plasmid from which an RNA was transcribed that coded for the third strand of a potential triple helix. A plasmid encoding the oligopurine variant of the triple helix but not the oligopyrimidine or a control sequence caused a dramatic reduction of IGF-I RNA and protein levels in cultured cells, morphological changes, and increased expression of protease nexin I and MHC class I molecules; the transfected cells displayed a reduced capacity for tumor growth when injected in nude mice (Shevelev et al, 1997). XXVII. Gene transfer to some characteristic tissues or cell types A. Transduction of hematopoietic stem cells (HSCs) Hematopoietic stem cells (HSCs), which can be isolated with high speed flow-cytometric cell sorting from fetal or adult bone marrow and cytokine-mobilized peripheral blood, have extensive self renewal and multilineage repopulating potential; HSCs are being used as an hematopoietic graft to treat cancer patients undergoing high dose chemotherapy which eradicates HSCs; GM-CSF treatment of the patient can enhance mobilization of true HSCs; furthermore, HSCs can be stably transduced at high efficiency (32-75%) by coculture with a cell line producing recombinant retroviruses containing the neomycin-resistant gene and are targets for hematopoietic cell-based gene therapy especially for the treatment of patients with multiple myeloma (Chen et al, 1995). The efficiency of gene transfer into monkey pluripotent hematopoietic stem cells (PHSCs) is at least one order of magnitude lower than what has been achieved in mice because primate PHSCs seem to require quite different culture conditions for their maintenance and transduction than mouse PHSCs. Successful retroviral vector-mediated gene transfer into monkey PHSCs supported maintenance of the long-term repopulating ability of autologous monkey grafts and has closed the gap between gene transfer experiments in mouse models and primates opening the door to the clinical application of bone marrow gene therapy to humans (Van Beusechem and Valerio, 1996). Retroviral vectors pseudotyped with vesicular stomatitis G glycoprotein (VSV-G) and expressing a murine cell surface protein, B7-1, were used to infect the human T-cell line Jurkat and human peripheral blood lymphocytes (PBLs); the transduction efficiency of PBLs with the pseudotyped vector reached a maximum of 16- 32% at an moi of 40 (Sharma et al, 1996). Introduction of a mutant H-ras gene (along with a neomycin resistance gene) into normal human bone marrow progenitor cells with a retrovirus followed by selection in cell culture with G418 suggested that expression of mutant H12-ras
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: Gene Therapy and Molecular Biology
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 41 and 42: C. Considerations of chromatin stru
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: C. Antisense ras gene transfer for
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 95 and 96: designed by immunization with porti
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

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?