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039 and MMS350 suppressed NFKb (≤ -38%), p21 (≤ -65%), and TNFa (≤ -99%), and elevated PGC-1A (≥
283%) (p ≤ 0.015). However, in wild-type Fancd2+/+ cells, JP4-039 and MMS350 induced NFKb (≥ 103%),
TGFb (≥ 98%), SP1 (≥ 181%), Gadd45 (≥ 392%), MnSOD (≥ 35%), Nrf2 (≥ 88%), and p21 (≥ 106%) (p ≤ 0.017).
The effects of JP4-039 on Fancd2-/- cells included decreased TGFb (-25%) and MnSOD (-40%), (p ≤ 0.021
for all groups). In contrast, MMS350 induced TGFb (+30%), SP1 (+68%), and Gadd45 (+239%) (p ≤ 0.022)
in Fancd2-/- cells. JP4-039 induced AP1 (+588%), and TNFa (+238%) (p ≤ 0.017) in wild type cells, whereas
MMS350 suppressed AP1 (-17%) and induced PGC-1A (+134%) (p ≤ 0.029) Conclusions: Radiation
mitigator drugs JP4-039 and MMS350 have similar biologic effects on radiosensitive cells but different
effects on RNA transcription, suggesting modulation of different molecular pathways. Supported by
NIAID/NIH U19-A 1068021 and the Fanconi Anemia Research Foundation.
(PS2-43) Doxycycline alters Total Body Irradiation (TBI) Induction of RNA transcripts in C57BL/6NHsd
Mice. Byung Han Rhieu; Ashwin Shinde; Michael Epperly; Tracy Dixon; Darcy Franicola; Julie Goff; Xichen
Zhang; Hong Wang; and Joel S. Greenberger, University of Pittsburgh Cancer Institute, Pittsburgh, PA
Purpose/Objectives: Doxycycline (doxy) induces MnSOD in tet/tet mice (Epperly et al; Radiation
Research 2013; 180(2):189-204) and is itself a radioprotector (Kim et al, Clinical Cancer Research, 2009
15(23):7238-45). We evaluated organ RNA transcript responses in doxy treated TBI C57BL/6NHsd (B6)
mice.
Materials/Methods: B6 mice (n=10) continuously received doxy in food for 2 weeks, then 9.25 Gy TBI.
Nineteen days later, mice were sacrificed, tissues harvested, RNA extracted, and transcript levels
quantitated by real time (rt) PCR, (n=4 mice/group), expressed as % change over baseline. Results: Control
mice at day 19 after TBI showed decreased MnSOD transcript levels in brain (-23.18±5.93, p=0.0211) and
kidney (-24.78±4.65, p=0.0018), doxy normalized to baseline levels (brain (-3.17±8.26, p=0.5745), kidney
(-15.25±15.21, p=0.1387)). MnSOD in the brains of doxy treated irradiated mice were significantly higher
compared to untreated irradiated mice (p=0.0271). GPX1 was increased in brains of control TBI mice
(24.41±1.56, p=0.0014), and doxy decreased it back to baseline levels (11.69±5.58, p=0.0682). GPX1 was
decreased in lungs of untreated irradiated mice (-13.82±2.55, p=0.0111), and doxy increased it back to
baseline levels (-8.92±10.92, p=0.2925). Nrf2 was upregulated in lungs (17.79±6.12, p=0.0373) of
untreated TBI mice, doxy normalized to baseline (lung 18.49±9.45, p=0.0772). Both doxy treated and
control TBI mice showed decreased NFkB in heart (-23.41±5.62, p=0.0036, and -23.17±9.93, p=0.0186,
respectively) and lung (-16.88±4.76, p=0.0255, and -17.19±6.52, p=0.0447, respectively), not different
from control TBI. TGFb was increased in brains and livers of control TBI mice (brain 23.42±2.73, p=0.0045,
liver 116.47±43.60, p=0.0437), doxy returned to baseline (brain 4.89±3.61, p=0.1435, liver 73.93±35.09,
p=0.0676). The decreased TGFb in brains of doxy treated TBI mice was lower compared to control TBI
(p=0.0021). Bone marrow of doxy treated TBI mice showed decreased (57.60±16.41, p=0.0260),
significantly lower than control TBI (p=0.0208). Doxy treated TBI mice had increased FAS in the brain
(59.73±10.99, p=0.0111). Conclusions: Doxycycline alters TBI induced mouse ¬RNA transcripts in vivo,
most prominent in brain.
158 | P a g e
283%) (p ≤ 0.015). However, in wild-type Fancd2+/+ cells, JP4-039 and MMS350 induced NFKb (≥ 103%),
TGFb (≥ 98%), SP1 (≥ 181%), Gadd45 (≥ 392%), MnSOD (≥ 35%), Nrf2 (≥ 88%), and p21 (≥ 106%) (p ≤ 0.017).
The effects of JP4-039 on Fancd2-/- cells included decreased TGFb (-25%) and MnSOD (-40%), (p ≤ 0.021
for all groups). In contrast, MMS350 induced TGFb (+30%), SP1 (+68%), and Gadd45 (+239%) (p ≤ 0.022)
in Fancd2-/- cells. JP4-039 induced AP1 (+588%), and TNFa (+238%) (p ≤ 0.017) in wild type cells, whereas
MMS350 suppressed AP1 (-17%) and induced PGC-1A (+134%) (p ≤ 0.029) Conclusions: Radiation
mitigator drugs JP4-039 and MMS350 have similar biologic effects on radiosensitive cells but different
effects on RNA transcription, suggesting modulation of different molecular pathways. Supported by
NIAID/NIH U19-A 1068021 and the Fanconi Anemia Research Foundation.
(PS2-43) Doxycycline alters Total Body Irradiation (TBI) Induction of RNA transcripts in C57BL/6NHsd
Mice. Byung Han Rhieu; Ashwin Shinde; Michael Epperly; Tracy Dixon; Darcy Franicola; Julie Goff; Xichen
Zhang; Hong Wang; and Joel S. Greenberger, University of Pittsburgh Cancer Institute, Pittsburgh, PA
Purpose/Objectives: Doxycycline (doxy) induces MnSOD in tet/tet mice (Epperly et al; Radiation
Research 2013; 180(2):189-204) and is itself a radioprotector (Kim et al, Clinical Cancer Research, 2009
15(23):7238-45). We evaluated organ RNA transcript responses in doxy treated TBI C57BL/6NHsd (B6)
mice.
Materials/Methods: B6 mice (n=10) continuously received doxy in food for 2 weeks, then 9.25 Gy TBI.
Nineteen days later, mice were sacrificed, tissues harvested, RNA extracted, and transcript levels
quantitated by real time (rt) PCR, (n=4 mice/group), expressed as % change over baseline. Results: Control
mice at day 19 after TBI showed decreased MnSOD transcript levels in brain (-23.18±5.93, p=0.0211) and
kidney (-24.78±4.65, p=0.0018), doxy normalized to baseline levels (brain (-3.17±8.26, p=0.5745), kidney
(-15.25±15.21, p=0.1387)). MnSOD in the brains of doxy treated irradiated mice were significantly higher
compared to untreated irradiated mice (p=0.0271). GPX1 was increased in brains of control TBI mice
(24.41±1.56, p=0.0014), and doxy decreased it back to baseline levels (11.69±5.58, p=0.0682). GPX1 was
decreased in lungs of untreated irradiated mice (-13.82±2.55, p=0.0111), and doxy increased it back to
baseline levels (-8.92±10.92, p=0.2925). Nrf2 was upregulated in lungs (17.79±6.12, p=0.0373) of
untreated TBI mice, doxy normalized to baseline (lung 18.49±9.45, p=0.0772). Both doxy treated and
control TBI mice showed decreased NFkB in heart (-23.41±5.62, p=0.0036, and -23.17±9.93, p=0.0186,
respectively) and lung (-16.88±4.76, p=0.0255, and -17.19±6.52, p=0.0447, respectively), not different
from control TBI. TGFb was increased in brains and livers of control TBI mice (brain 23.42±2.73, p=0.0045,
liver 116.47±43.60, p=0.0437), doxy returned to baseline (brain 4.89±3.61, p=0.1435, liver 73.93±35.09,
p=0.0676). The decreased TGFb in brains of doxy treated TBI mice was lower compared to control TBI
(p=0.0021). Bone marrow of doxy treated TBI mice showed decreased (57.60±16.41, p=0.0260),
significantly lower than control TBI (p=0.0208). Doxy treated TBI mice had increased FAS in the brain
(59.73±10.99, p=0.0111). Conclusions: Doxycycline alters TBI induced mouse ¬RNA transcripts in vivo,
most prominent in brain.
158 | P a g e
