Page 245 - 2014 Printable Abstract Book
P. 245
treatment. CM-H2DCFDA staining indicated that R71 regulated intracellular ROS production. Minimally
toxic doses of R71 increased sensitivity to radiation and cisplatin. Conclusion: R71 has antiproliferative
effects with an IC50 in the μM range. The mechanism likely includes apoptosis associated with ROS
production and interruption of mitochondrial membrane potential. A549 cells were also sensitized to
radiation and cisplatin treatment.
(PS4-18) Antibody targeting of a radiation inducible tax-interaction protein 1 (tip 1) as a novel molecule
for tumor treatment. Heping Yan, MD; Kim Nguyen, Ph.D; Vaishali Kapoor, Ph.D; Steve Mnich; Hua Li,
Ph.D; Buck Rogers, Ph.D; Dinesh Thotala, Ph.D; and Dennis Hallahan* Corresponding author, MD
Washington University in St. Louis, St. Louis, MO
Ionizing radiation (IR) can achieve cell killing and elicit phenotype changes in tumor cells resulting
in molecules being expressed on the cell surface which can be exploited as targets for tumor imaging and
(1)
therapy. We observed Tip1 to be an IR inducible protein . Here we report that several clones of anti-Tip
1 Mabs have been developed and characterized which can be used to deliver anti-tumor drugs or radio-
isotopes to cancer. An IgG2b anti-Tip 1 Mab, 2C6F3, has been chosen for further studies. The ELISA and
IHC data indicated this Mab had high specificity and affinity to Tip 1. It recognizes a ~14 kDa protein
corresponding to Tip 1 by W-B. Tip1 expression increased at 8 and 24 hours after IR treatment in a panel
of tumor cell lysate of LLC, D54, A549, PC-3, GL261. Further, IR increased Tip 1 expression on cultured
tumor cell surface. Mab binds to tumor cells of LLC, D54, A549 and H460 as determined by IF and FACS.
A549 cells showed approximately 4 fold increases in Tip1 surface expression and 2 fold increases in D54
cells. The level of Tip1 expressed on H460 cells surface increased 24 hours after IR and is dose dependent
(1)
with 2Gy, 4Gy and up to 8Gy compared with 0Gy treatment . Purified Mab was conjugated to AF750 and
injected into LLC and GL261 tumor bearing mice exposed to either 3Gy x 3 or sham 0Gy treatment. The
signal of labeled Mab bound to IRed tumor was measured. The intensity increased after 24 hours and the
signal duration is at least one week. Tumor with 0Gy had minor binding and diminished within 24-48
hours. The control NMIgG showed no tumor binding over entire course of imaging. Conjugating the Mab
90
64
to radio-isotope of Cu, 111 In, Y, 125 I was effective for in vivo imaging. The labeling specificity and
efficiency was evaluated by ELISA and thin layer romatography. The radio-isotope conjugated Mab was
injected into GL261 and LLC tumor bearing mice for in vivo distribution and micro PET/CT or nano-
CT/SPECT imaging. We found that this Mab bound to 3Gy treated GL261 tumors while 0Gy had minimal
binding. SPECT imaging showed enhanced tumor binding of 111 In and 125 I labeled Ab on irradiated LLC
111
tumor in C57/BL at 48 hours. The distribution data reveled high binding of In - Ab after 24-48 hours post
90
IR. Also, a dose dependent assay was performed with Y conjugated 2C6F3 showed 300μCi of labeled
Mab had higher binding on A549 tumor bearing mice.
(PS4-19) The radical chemistry of the anticancer triazine 1, 4-dioxide hypoxia-activated prodrug,
SN30000. Robert F. Anderson, PhD; Pooja Yadav, PhD; and Michael Hay, PhD, University of Auckland,
Auckland, New Zealand
The radical chemistry underlying the activity of the bioreductive anticancer benzotriazine 1,4-
dioxide (BTO) prodrug, SN30000, which is soon to enter clinical trials, has been investigated by pulse
radiolysis (PR) and electron paramagnetic resonance (EPR) techniques. Upon one-electron reduction, the
243 | P a g e
toxic doses of R71 increased sensitivity to radiation and cisplatin. Conclusion: R71 has antiproliferative
effects with an IC50 in the μM range. The mechanism likely includes apoptosis associated with ROS
production and interruption of mitochondrial membrane potential. A549 cells were also sensitized to
radiation and cisplatin treatment.
(PS4-18) Antibody targeting of a radiation inducible tax-interaction protein 1 (tip 1) as a novel molecule
for tumor treatment. Heping Yan, MD; Kim Nguyen, Ph.D; Vaishali Kapoor, Ph.D; Steve Mnich; Hua Li,
Ph.D; Buck Rogers, Ph.D; Dinesh Thotala, Ph.D; and Dennis Hallahan* Corresponding author, MD
Washington University in St. Louis, St. Louis, MO
Ionizing radiation (IR) can achieve cell killing and elicit phenotype changes in tumor cells resulting
in molecules being expressed on the cell surface which can be exploited as targets for tumor imaging and
(1)
therapy. We observed Tip1 to be an IR inducible protein . Here we report that several clones of anti-Tip
1 Mabs have been developed and characterized which can be used to deliver anti-tumor drugs or radio-
isotopes to cancer. An IgG2b anti-Tip 1 Mab, 2C6F3, has been chosen for further studies. The ELISA and
IHC data indicated this Mab had high specificity and affinity to Tip 1. It recognizes a ~14 kDa protein
corresponding to Tip 1 by W-B. Tip1 expression increased at 8 and 24 hours after IR treatment in a panel
of tumor cell lysate of LLC, D54, A549, PC-3, GL261. Further, IR increased Tip 1 expression on cultured
tumor cell surface. Mab binds to tumor cells of LLC, D54, A549 and H460 as determined by IF and FACS.
A549 cells showed approximately 4 fold increases in Tip1 surface expression and 2 fold increases in D54
cells. The level of Tip1 expressed on H460 cells surface increased 24 hours after IR and is dose dependent
(1)
with 2Gy, 4Gy and up to 8Gy compared with 0Gy treatment . Purified Mab was conjugated to AF750 and
injected into LLC and GL261 tumor bearing mice exposed to either 3Gy x 3 or sham 0Gy treatment. The
signal of labeled Mab bound to IRed tumor was measured. The intensity increased after 24 hours and the
signal duration is at least one week. Tumor with 0Gy had minor binding and diminished within 24-48
hours. The control NMIgG showed no tumor binding over entire course of imaging. Conjugating the Mab
90
64
to radio-isotope of Cu, 111 In, Y, 125 I was effective for in vivo imaging. The labeling specificity and
efficiency was evaluated by ELISA and thin layer romatography. The radio-isotope conjugated Mab was
injected into GL261 and LLC tumor bearing mice for in vivo distribution and micro PET/CT or nano-
CT/SPECT imaging. We found that this Mab bound to 3Gy treated GL261 tumors while 0Gy had minimal
binding. SPECT imaging showed enhanced tumor binding of 111 In and 125 I labeled Ab on irradiated LLC
111
tumor in C57/BL at 48 hours. The distribution data reveled high binding of In - Ab after 24-48 hours post
90
IR. Also, a dose dependent assay was performed with Y conjugated 2C6F3 showed 300μCi of labeled
Mab had higher binding on A549 tumor bearing mice.
(PS4-19) The radical chemistry of the anticancer triazine 1, 4-dioxide hypoxia-activated prodrug,
SN30000. Robert F. Anderson, PhD; Pooja Yadav, PhD; and Michael Hay, PhD, University of Auckland,
Auckland, New Zealand
The radical chemistry underlying the activity of the bioreductive anticancer benzotriazine 1,4-
dioxide (BTO) prodrug, SN30000, which is soon to enter clinical trials, has been investigated by pulse
radiolysis (PR) and electron paramagnetic resonance (EPR) techniques. Upon one-electron reduction, the
243 | P a g e
