GTMB 7 - Gene Therapy & Molecular Biology

More documents

Recommendations

Info

Epperly et al: Late injection of MnSOD-PL protects against pulmonary fibrosisTherefore, MnSOD-PL action on ROS produced byinflammatory cells such as macrophages at 80 days doesnot appear to explain the present data implyingsuperoxides might have been produced by macrophagesand neutralized by injections of MnSOD-PL at 80 or 100days after irradiation (Epperly et al, 2003).We previously demonstrated that at 80 days afterirradiation of the mouse lung there is an increase in TNF-αmRNA expression which decreases to background level byday 120 (Epperly et al, 1999b). This increase isaccompanied by increased expression of mRNA for TGFβat day 100. Initially, there is an increase in TGF-β1isoform until day 120 at which time TGF-β1 expressiondecreases, and TGF-β2 expression increases and stayselevated until development of organizing alveolitis/fibrosis(Epperly et al, 1999b). The detectable protection byMnSOD-PL injection at day 80 might have been attributedto an effect on the TNF-α elevation at that time point.ROS production might increase TNF-α expression at day80 leading to further increased ROS production (Haddad,2002). Treatment of alveolar epithelial cells with a ROSgenerating system results in increased TNF-α expressionand a depletion of glutathione (Haddad, 2002). TNF-αtreatment inhibits myogenesis by causing a decrease inglutathione levels and elevation of ROS (Langen et al,2002). Pre-treatment with the anti-oxidant N-acetyl-1-cysteine (NAC) restored the formation of multi-nucleatedmyotubes, indicating that myogenesis inhibition wasattributable to ROS expression (Langen et al, 2002). Pretreatmentof HELA cells with gammaglutamylcysteinylglycineinhibits TRAIL-inducedapoptosis (Lee et al, 2002). TNF-α expression mayincrease the production of ROS and result in a furtherincrease in TNF-α expression. ROS response to andinduction of TNF-α expression may be a cyclicmechanism in the lung at day 80, and MnSOD-PLtreatment at this time point may have interrupted the cycle.Further studies will be required to explain the protectionby injections of MnSOD-PL at 80 days after irradiation.Pulmonary increases in TGF-β1 and TGF-β2 mRNAat 100 to 120 days after irradiation have been detected(Epperly et al, 1999b). It has been demonstrated that ROScan also increase TGF-β expression (Bellocq et al, 1999)The treatment of human alveolar lung cell line A549 withxanthine and xanthine oxidase or nitric oxide generator S-nitroso-N-acetyl-penicillamine (SNAP) leads to release ofTGF-β1 (Bellocq et al, 1999). The xanthine-xanthineoxidase induced release of TGF-β1 can be inhibited by theaddition of catalase but not superoxide dismutase,implicating the involvement of hydrogen peroxide(Bellocq et al, 1999) TGF-β1 has been demonstrated toinduce production of extracellular hydrogen peroxide inhuman fibroblasts that mediate oxidative dityrosinedependentcross-linking of ECM (Larios et al, 2001).TGF-β and hydrogen peroxide have been observed toinduce connective tissue factor (CTGF) that then inducescollagen type 1 and fibronectin, a deposition leading tofibrosis (Park et al, 2001).The mechanism of action of TGF-β in cells of thelung may involve the mitochondria since TGF-β1 can leadto downregulation of Bcl-2 and Bcl-xl, which normallyprevent apoptosis (Lafon et al, 1996; Herrera et al, 2001a).Overexpression of Bcl-2 suppresses the effects of TGF-β(Huang and Chou, 1998). Following exposure to TGF-βthere is also a loss of mitochondrial membrane potential,release of cytochrome-C, and activation of caspase-3(Herrera et al, 2001a). TGF-β1 activates caspase- 3, 8 and9, which precede the loss of mitochondrial membranepotential (Herrera et al, 2001b). Activation of caspase-8results in cleavage of Bid and Bcl-xl, which may lead toan amplification loop resulting in the mitochondrialmediated apoptosis (Zha et al, 2000). Irradiation of murinebone marrow stromal cell line D2XRII in vitro inducesrelease of TGF-β into the culture medium (Greenberger etal, 1996). Co-cultivation of 32D cl 3 cells or subclones1F2 or 2C6 overexpressing MnSOD with irradiated bonemarrow stromal cells resulted in higher levels ofintracellular ROS in the non-irradiated 32D cl 3, 1F2 or2C6 cells compared to cells co-cultivated with nonirradiatedstromal cell lines (Greenberger et al, 1996). TheMnSOD overexpressing subclonal line 1F2 or 2C6 formedmore cobblestone islands on the irradiated stromal cellsthan 32D cl 3 cells (Greenberger et al, 1996). IncreasedMnSOD activity in 1F2 or 2C6 cells may have resulted ina decrease in ROS, thus allowing for greater attachment ofthe MnSOD overexpressing cell lines to the irradiatedstromal cells. Therefore, injections of MnSOD-PL into thelung at day 100 when TGF-β levels are beginning toincrease may inhibit ROS production, and/or stabilize thebronchoalveolar cell or endothelial cell mitochondria,preventing some (but not all) of the late effects ofirradiation damage to the lung. We are currently exploringthis possible mechanism.The present report indicates that a singleadministration of MnSOD-PL 24 hours prior to 20 Gylung irradiation is significantly more effective thanadministration at any of four post-irradiation time pointsranging from 1-100 days after irradiation. We did notevaluate time points between 1 and 80 days as there wasno histopathologic or other evidence to suggest thatinitiation steps in the late organizing alveolitis/fibrosisresponse began prior to 80 days. Our results may helpexplain the dynamics of late irradiation pulmonary injury.One interpretation of the results is that it representsevidence of a pleiotropic effect of ionizing irradiation onseveral cellular and physiologic targets within the lung.Initiation events at the time of irradiation may lead to amultiplicity of effectuating events beginning at around day100 and leading to rapid organizing alveolitis/fibrosis.Prevention of some of the initiating events by MnSOD-PLadministration prior to irradiation may have a significantlygreater effect at reducing the overall outcome compared tomodulation of some of the late effectuating events byMnSOD-PL administration at that time. For example,neutralization of free radical moieties induced byirradiation at day 0 by overexpression of MnSOD at thattime may be a significant early event which impacts onmultiple downstream/delayed effectuating targets(macrophage migration, fibroblast migration into the lung,66
Gene Therapy and Molecular Biology Vol 7, page 67endothelial upregulation of adhesion molecules, and othercomponents of the fibrosis response not yet elucidated). Incontrast, modulation of some of the effectuating events byMnSOD-PL administration at the late time points mayhave a significantly decreased effect in preventing latelesion simply due to the multiplicity of events already inprogress, and that many of these may be unrelated to thefree radical neutralization capacity of MnSOD at that latetime point. The same mechanism explaining a greatereffect of treatment prior to irradiation might also hold truefor anti-apoptotic effects of MnSOD overexpression in themitochondria.The present data also indicate that significantprotective effects afforded by MnSOD-PL administrationprior to irradiation were not significantly further improvedby a second delayed administration. This result maysimply be attributable to the dominant mechanism ofprevention of initiating events compared to effectuatingevents. The present results add support to utilization offractionated inhalation of freeze-dried MnSOD-PL duringcourses of fractionated radiotherapy which would beappropriate to the clinical translational model of normallung irradiation protection in lung cancer patientsreceiving chemoradiotherapy over a 60-day time course.Fractionation experiments currently in progressincorporate twice weekly inhalation of freeze-driedMnSOD-PL by mice receiving 24 fractions of irradiationduring 35 days. The present observation of a lack oftoxicity of a second delayed administration of MnSOD-PLin the present data supports the concept that multi-fractionadministration of this gene therapy technique should notexacerbate and may decrease pulmonary irradiationdamage.AcknowledgmentsThis research has been supported by the NationalInstitutes of Health, Grant #R01-HL-60132ReferencesBellocq A, Azoulay E, Marullo S, Flahault A, Fouqueray B,Philippe C, Cadranel J, Baud L (1999) Reactive oxygen andnitrogen intermediates increase transforming growth factorbeta1 release from human epithelial alveolar cells throughtwo different mechanisms. AJRCMB 21, 128-136.Bowler RP, Crapo JD (2002) Oxidative stress in airways, is therea role for extracellular superoxide dismutase? AJRCCM166, 38-43.Epperly MW, Bray JA, Kraeger S, Swacka R, Engelhardt JF,Travis E, and Greenberger (1998) Prevention of late effectsof irradiation lung damage by manganese superoxidedismutase gene therapy. Gene Ther 5, 196-208.Epperly MW, Bray JA, Esocobar P, Bigbee WL, Watkins S,Greenberger JS (1999a) Overexpression of the humanMnSOD transgene in subclones of murine hematopoieticprogenitor cell line 32D cl 3 decreases irradiation-inducedapoptosis but does not alter G2/M or G1/S phase cell cyclearrest. Radiat Oncol Invest 7, 331-342.Epperly, MW, Bray, JA, Krager, S, Berry, LM, Gooding, W,Engelhardt, JF, Zwacka, R, Travis,EL, and Greenberger, JS(1999b) Intratracheal injection of adenovirus containing thehuman MnSOD transgene protects athymic nude mice fromirradiation-induced organizing alveolitis. Int J Radiat OncolPhys 43, 169-181.Epperly MW, Travis EL, Sikora C, Greenberger JS (1999c)Magnesium superoxide dismutase (MnSOD)plasmid/liposome pulmonary radioprotective gene therapy,Modulation of irradiation-induced mRNA for IL-1, TNF-α,and TGF-β correlates with delay of organizingalveolitis/fibrosis. Biol Blood Bone Marrow Transplant 5,204-214.Epperly MW, Epstein CJ, Travis EL, Greenberger JS (2000a)Decreased pulmonary radiation resistance of manganesesuperoxide dismutase (MnSOD)-deficient mice is correctedby human manganese superoxide dismutaseplasmid/liposome(SOD2-PL) intratracheal gene therapy.Radiat Res 154, 365-374.Epperly MW, Sikora C, Defilippi S, Bray J, Koe G, Liggitt D,Luketich JD, Greenberger JS (2000b) Plasmid/liposometransfer of the human manganese superoxide dismutase(MnSOD) transgene prevents ionizing irradiation-inducedapoptosis in human esophagus organ explant culture. RadiatOncol Invest 90, 128-137.Epperly MW, Gretton JA, DeFilippi SJ, Greenberger JS, SikoraCA, Liggitt D, Koe G (2001a) Modulation of radiationinducedcytokine elevation associated with esophagitis andesophageal stricture by manganese superoxide dismutaseplasmid/liposome(SOD-PL) gene therapy. Radiat Res 155,2-14.Epperly MW, Travis EL, Whitsett JA, Raineri I, Epstein CJ,Greenberger JS (2001b) Overexpression of manganesesuperoxide dismutase (MnSOD) in whole lung or alveolartype II (AT-II) cells of MnSOD transgenic mice does notprovide intrinsic lung irradiation protection. Radiat OncolInvest 96, 11-21.Epperly MW, Guo HL, Gretton JE, Greenberger JS. (2003) Bonemarrow origin of myofibroblasts in irradiation pulmonaryfibrosis. AJRCMB, 29, in press.Epperly MW, Sikora CA, DeFilippi SJ, Gretton JA, Zhan Q,Kufe DW, Greenberger JS (2002) MnSOD inhibitsirradiation-induced apoptosis by stabilization of themitochondrial membrane against the effects of SAP kinasesp38 and Jnk1 translocation. Radiat Res 157, 568-577.Fridovich I (1995) Superoxide radical and superoxidedismutases. Annu Rev Biochem 64, 97-112.Gorbunov NV, Pogue-Geile KL, Epperly MW, Bigbee WL,Draviam R, Day BW, Wald N, Watkins SC, Greenberger JS(2000) Activation of the nitric oxide synthase 2 pathway inthe response of bone marrow stromal cells to high doses ofionizing radiation. Radiat Res 154, 73-86.Gossart S, Cambon C, Orfila C, Seguelas MH, Lepert JC, RamiJ, Carre P, Pipy B (1996) Reactive oxygen intermediates aregulators of TNF-alpha production in rat lung inflammationinduced by silica. J Immun 156, 1540-1548.Greenberger JS, Epperly MW, Zeevi A, Brunson KW, GoltryKL, Pogue-Geile KL, Bray J, Berry L (1996) Stromal cellinvolvement in leukemogenesis and carcinogenesis. In Vivo10, 1-18.Guo HL, et al. (2003) Prevention of irradiation-induced oralcavity mucositis by plasmid/liposome delivery of the humanmanganese superoxide dismutase (MnSOD) transgene.Radiat Res 159, 361-370.Haddad JJ (2002) Redox regulation of pro-inflammatorycytokines and IkappaB-alpha/NF-kappaB nuclear67
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: Gene Therapy and Molecular Biology
Page 77: Gene Therapy and Molecular Biology
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 130 and 131:
Sanlioglu et al: Adenovirus mediate
Page 132 and 133:
Page 134 and 135:
Page 136 and 137:
Page 138 and 139:
Page 140 and 141:
Page 142 and 143:
Page 144 and 145:
Page 146 and 147:
Page 148 and 149:
Page 150 and 151:
George et al: Gene therapy for vasc
Page 152 and 153:
Page 154 and 155:
Page 156 and 157:
Page 158 and 159:
Page 160 and 161:
Page 162 and 163:
Page 164 and 165:
Page 166 and 167:
Page 168 and 169:
Zhang et al: Angiogenic Gene Therap
Page 170 and 171:
Page 172 and 173:
Page 174 and 175:
Page 176 and 177:
Page 178 and 179:
Page 180 and 181:
Page 182 and 183:
Xu et al: G-CSF receptor-mediated S
Page 184 and 185:
Page 186 and 187:
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

Create successful ePaper yourself

Delete template?

Save as template?