GTMB 7 - Gene Therapy & Molecular Biology

More documents

Recommendations

Info

Sanlioglu et al: Adenovirus mediated gene therapy for prostate carcinomaactivity exhibit TRAIL resistance (Franco et al, 2001).Resistant melanoma cells were sensitized to TRAIL eitherwith proteasome inhibitors or transfections with plasmidsencoding degradation resistant IκBα protein (Franco et al,2001). In accordance with these studies, we have tested ifadenovirus mediated NF-κB inhibiting approach wouldsensitize prostate cancer cells to TRAIL. Consequently,adenovirus mediated delivery of IKKβKA mutant(Ad.IKKβKA) sensitized PTEN mutant prostate cancercells (PC3) to TRAIL as shown in Figure 2. At first, PC3cells appeared to be relatively resistant to pro-apoptoticeffects of TRAIL when cells were infected withadenovirus vector encoding hTRAIL (Ad.hTRAIL) evenat an MOI of 1000 DNA particles/cell (Figure 2 Panel A).Infection with Ad.IKKβKA vector alone did not yield anycell death either (Figure 2, Panel B). However, when thedose of Ad.hTRAIL vector was kept constant at an MOI of1000 DNA particles/cell, increasing the amount ofAd.IKKβKA construct sensitized PC3 cells to TRAILmediated apoptosis (Figure 2, Panel C).C. Intracellular proapoptotic regulatorsAlthough caspases are the effector mediators of apoptosis,the expression of proapoptotic molecules such asprocaspase 3 or 7 using adenovirus constructs did notinduce apoptosis in prostate cancer cells due to theinability of these caspases to undergo autocatalyticactivation (Li et al, 2001). A novel suicide gene therapyapproach was developed using chemically inducibleeffector caspases to trigger apoptosis in prostate cancercells. Cell death was mediated by replication-deficientadenoviral vector expressing conditional caspase-1 (Ad-G/iCasp1) or caspase-3 (Ad-G/iCasp3) and the caspaseactivation was achieved by nontoxic, lipid-permeable,chemical inducers of dimerization (CID) (Shariat et al,2001). Aggregation and activation of these recombinantcaspases occurred, leading to rapid apoptosis only aftervector transduction followed by CID administration inboth human (LNCaP and PC-3) and murine (TRAMP-C2and TRAMP-C2G) prostate cancer cell lines.Subcutaneous TRAMP-C2 tumors displayed focal butextensive apoptosis following direct injection of Ad-G/iCasp1 in vivo. In order to express caspase 9exclusively in prostate, a recombinant adenovirus carryingiCaspase-9 was constructed with two copies of theandrogen response region (ARR) placed upstream of theprobasin promoter elements (ADV.ARR(2)PB-iCasp9)(Xie et al, 2001b). AP20187 is a chemical dimeric ligand,which causes dimerization and thereby activation ofiCaspase-9 leading to rapid apoptosis in both dividing andnondividing cells. Testing of ADV.ARR(2)PB-iCasp9construct in LNCaP tumor xenografts demonstrated thatthis construct induces apoptosis in prostate cancer cellsonly in the presence of AP20187.The proapoptotic members of Bcl- 2 protein familyincluding Bax, Bak, Bad, and Bik also mediate apoptosis.Apoptosis-inducing proteins were cloned into adenovirusconstructs and shown to induce apoptosis in prostatecancer cell lines previously.Figure 2. Adenovirus mediated IKKβKA expression sensitized PC3 cells to TRAIL mediated apoptosis. PC3 cells were infected withincreasing MOIs of either Ad5hTRAIL (Panel A) or Ad.IKKβKA (Panel B). In panel C, the dose of Ad.IKKβKA vector was increasedgradually (stated just above each panel) while the amount of Ad5hTRAIL was kept constant (as indicated under the panel). Cell deathwas detected using molecular probe’s Live and Death Cellular viability and toxicity kit 48 hours following infection. Numbers indicateviral doses as MOI values of DNA particles/cell.120
Gene Therapy and Molecular Biology Vol 7, page 121However, overexpression of proapoptotic genes withoutthe use of tissue specific promoters could result inunwanted apoptosis even in normal cells. In order toprovide tissue specificity, an adenoviral construct wasgenerated containing Bax cDNA under control of theprobasin promoter that included two androgen responseelements (Av-ARR2PB-Bax). Av-ARR2PB-Bax constructdrove Bax overexpression in an androgen-dependent wayin androgen receptor (AR)-positive cell lines of prostaticorigin but not in others. The androgen dihydrotestosteroneactivated apoptosis in LNCaP cells infected with Av-ARR2PB-Bax but not in those infected with controlvectors. These results demonstrated that Av-ARR2PB-Baxinduced apoptosis was androgen dependent and limited toAR positive cells of prostatic epithelium. On the otherhand, using a binary co-transfection strategy involvingAd/GT Bax and Ad/PGK-GV16; overexpression ofproapoptotic Bax protein induced apoptosis both inandrogen-insensitive (DU145 and PC3), and androgensensitive(LNCaP) cell lines (Honda et al, 2002). The samebinary approach was tested to assess the consequences ofBcl-2 overexpression in the progression of prostatecarcinoma leading to apoptosis-resistant and androgenindependentphenotype in DU145, PC3 and LNCaP celllines which represent models of advanced prostatecarcinoma. Bax expression generated by the adenoviralco-transfection system induced apoptosis even in theseBcl-2 overexpressing cell lines. These results suggest thatthe Ad/GT Bax and Ad/PGK-GV16 combined expressionsystem might represent a powerful gene therapy strategyfor the treatment of androgen-independent and apoptosisresistantprostate carcinoma. Moreover, monogene andpolygene approaches were compared in an experimentalprostate cancer model using apoptotic genes bad and baxdriven by a prostate specific promoter (ARR(2)PB) in anadenovirus construct (Zhang et al, 2002b). The ARR(2)PBis a dihydrotestosterone (DHT)-inducible third-generationprobasin-derived promoter. In this study, animals bearingtumors of prostatic origin responded better to combinedbad and bax therapy than either of the vectors alone.Therefore, it was concluded that polygene therapyinvolving more than one apoptotic molecule is moreeffective in xenograft models of androgen-dependent orindependent prostate cancer than monogene therapy alone.It is also known that overexpression of anti-apoptoticgenes such as Bcl-2 in prostate carcinoma providesresistance to radiation therapy and androgen ablation. Asecond-generation adenoviral vector (ARR2PB.Bax.GFP)was constructed with the modified prostate-specificprobasin promoter (ARR2PB) directing the expression of aHA-tagged Bax gene in order to restore the balance ofBcl-2 family members to induce apoptosis in prostatecancer cells (Lowe et al, 2001). ARR2PB.Bax.GFP vectorinduced significant levels of apoptosis in LNCaP cells 48hours following infection even in the presence of highlevels of Bcl-2 protein. No toxicity in liver, lung, kidney,and spleen was detected by systemic administration ofARR2PB.Bax.GFP in nude mice. Therefore, a secondgenerationadenovirus-mediated, prostate-specific Baxgene therapy appeared to be a very safe and efficientapproach for the treatment of prostate cancer. Anothermember of the proapoptotic Bcl-2 family, namely "Bik",was cloned into adenovirus vectors to explore itstherapeutic potential. AdBik infection also inducedapoptosis and suppressed the growth of PC-3 xenograftsestablished in nude mice (Tong et al, 2001).Several other genes were also tested for their abilityto induce apoptosis in prostate tumor cell lines as well asin xenograft models. The antiapoptotic protein CLN3negatively regulates endogenous ceramide production, aninducer of apoptotic cell death. CLN3 protein isoverexpressed in most of the cancer cell lines testedincluding those of prostate (Du145, PC-3, and LNCaP).An adenovirus-expressing antisense CLN3 (Ad-AS-CLN3) blocked CLN3 protein expression in prostatecancer cell lines as demonstrated by Western Blotting(Rylova et al, 2002). Ad-AS-CLN3 infection resulted inthe inhibition of cell growth and reduction in cell viabilityof cancer cells through elevation of endogenous ceramideproduction. This study revealed CLN3 as a novel target toinduce apoptosis in prostate cancer cells. A recombinantadenovirus containing pHyde cDNA gene (AdpHyde), anovel gene cloned from Dunning rat prostate cancer cells,was constructed in order to study its function (Zhang et al,2001). Surprisingly, the AdpHyde construct inhibited thegrowth of human prostate cancer cells and inducedapoptosis involving the caspase-3 pathway in humanprostate cancer tumor xenografts in nude mice. Ionicmovement also influences apoptosis. For instance, K +efflux is an early event in apoptosis, which is regulated byK + channel-associated protein (KChAP). A recombinantadenovirus encoding KChAP (Ad/KChAP) wasconstructed in order to determine if KChAP expressioncould induce apoptosis in prostate cancer cells (Wible etal, 2002). The LNCaP cell line displayed a reduction incell size upon infection with Ad/KChAP. The Ad/KChAPconstruct also induced apoptosis in DU145 cells in a p53independent manner. In addition, infection withAd/KChAP prevented growth of DU145 and LNCaPtumor xenografts in nude mice.VII. Tumor suppressor genesAberrations in the expression of tumor suppressorgenes have been one of the key factors affecting theoutcome of cancer therapy. Several studies examined thepossible use of tumor suppressor genes as therapeuticagents for prostate cancer. Doxorubicin (Dx) is acommonly used chemotherapeutic agent in recurrentprostate cancer and is a strong inducer of p53 expressionleading to p21(CIP1/WAF1) transactivation. As suggestedby previous reports, p21 plays a role in the modulation ofchemotherapy-induced apoptosis, prostate cancerprogression and androgen regulation. Two androgenregulatedhuman prostate cancer cell lines (MDA PCa 2band LNCaP) were exposed to Dx and growth factorwithdrawal in order to investigate if p21 plays a role in thesurvival of prostate cancer cells under stress (Martinez etal, 2002). Infection with adenovirus vectors encoding theantisense strand of p21 reduced p21 levels, sensitizedprostate cancer cells to Dx and facilitated apoptosis inresponse to growth factor withdrawal. These resultssuggest that modulation of p21 pro-survival gene121
Page 7 and 8:
Instructions to authors:Gene Therap
Page 9:
Please submit an electronic version
Page 12:
103-111 ResearchArticle113-133 Revi
Page 17 and 18:
Gene Therapy and Molecular Biology
Page 19 and 20:
Page 21 and 22:
Page 23 and 24:
Page 25 and 26:
Page 27 and 28:
Page 29 and 30:
Page 31 and 32:
Page 33 and 34:
Page 35 and 36:
Page 37 and 38:
Page 39 and 40:
Page 41 and 42:
Page 43 and 44:
Page 45 and 46:
Page 47 and 48:
Page 49 and 50:
Page 51 and 52:
Page 53 and 54:
Page 55 and 56:
Page 57 and 58:
Page 59 and 60:
Page 61 and 62:
Page 63 and 64:
Page 65 and 66:
Page 67 and 68:
Page 69 and 70:
Page 71 and 72:
Page 73 and 74:
Page 75 and 76:
Page 77:
Page 80 and 81:
Epperly et al: Late injection of Mn
Page 82 and 83:
Epperly et al: Late injection of Mn
Page 84 and 85: Goldberg-Cohen et al: Regulation of
Page 90 and 91: Gascón-Irún et al: Gene therapy a
Page 106 and 107: Suzuki et al: Regulation of the Sp/
Page 114 and 115: Li et al: MET amplification in live
Page 116 and 117: Li et al: MET amplification in live
Page 118 and 119: Chavakis et al: Leukocyte adhesion
Page 128 and 129: Sanlioglu et al: Adenovirus mediate
Page 150 and 151: George et al: Gene therapy for vasc
Page 168 and 169: Zhang et al: Angiogenic Gene Therap
Page 182 and 183: Xu et al: G-CSF receptor-mediated S
Page 184 and 185:
Xu et al: G-CSF receptor-mediated S
Page 186 and 187:
Xu et al: G-CSF receptor-mediated S
Page 188 and 189:
Burek et al: Calcium induced cell d
Page 190 and 191:
Page 192 and 193:
Page 194 and 195:
Page 196 and 197:
David et al: Current status and fut
Page 198 and 199:
Page 200 and 201:
Page 202 and 203:
Page 204 and 205:
Page 206 and 207:
Page 208 and 209:
Page 210 and 211:
Page 212 and 213:
Page 214 and 215:
Page 216 and 217:
Page 218 and 219:
Page 220 and 221:
Page 222 and 223:
Page 224 and 225:
Page 226 and 227:
Stoll et al: The role of EBV and ge
Page 228 and 229:
Page 230 and 231:
Page 232 and 233:
Page 234 and 235:
Page 236 and 237:
Maruyama et al: Kidney-targeted pla
Page 238 and 239:
Page 240 and 241:
Page 242 and 243:
Page 244 and 245:
Kren et al: Hepatocyte-targeted del
Page 246 and 247:
Page 248 and 249:
Page 250 and 251:
Page 252 and 253:
Page 254 and 255:
Zeng: PRL-3 as a target for cancer
Page 256 and 257:
Page 258 and 259:
Page 260 and 261:
Latchman: Protective effect of heat
Page 262 and 263:
Page 264 and 265:
Page 266 and 267:
Page 268 and 269:
Page 270 and 271:
Cai et al: Lung cancer gene therapy
Page 272 and 273:
Page 274 and 275:
Page 276 and 277:
Page 278 and 279:
Page 280:
Page 283 and 284:
Page 285 and 286:
Page 287 and 288:
Page 289 and 290:
Page 291 and 292:
Page 293 and 294:
Page 295 and 296:
Page 297 and 298:
Page 299 and 300:
Page 301 and 302:
Page 303 and 304:
Page 305 and 306:
Page 307 and 308:
Page 309:
show all

GTMB 7 - Gene Therapy & Molecular Biology

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

Delete template?

Save as template?