Page 314 - 2014 Printable Abstract Book
P. 314
(PS5-47) Utilising archived tissues from mouse lifespan studies to characterise carbon-ion radiotherapy-
induced second tumours. Benjamin J. Blyth, PhD; Shizuko Kakinuma; Yutaka Yamada; Takamitsu Morioka;
Shinobu Hirano-Sakairi; Yoshiko Amasaki; Yi Shang; Chizuru Tsuruoka; Tatsuhiko Imaoka; Mayumi
Nishimura; and Yoshiya Shimada, National Institute of Radiological Sciences, Chiba, Japan
The National Institute of Radiological Sciences, Japan, utilises a range of experimental animal
models in both short-term and lifespan studies to investigate tumours across an array of tissue sites
induced by X-ray, γ-ray, neutron or heavy-ion exposure, focusing on carbon ion radiotherapy as performed
at the HIMAC facility (Heavy Ion Medical Accelerator in Chiba) at our institute in particular. We are in the
process of establishing ‘J-Share’, an online archive to provide access to experimental data from our
collection of lifespan/cancer incidence studies, to ultimately facilitate the world-wide sharing of data and
archived tissues for collaborative endeavours. Here, we report on one such analysis of archived samples
of T-cell lymphomas induced by whole-body exposure(s) of 1-week-old mice to a monoenergetic carbon
ion beam (13 keV) simulating the exposure of normal tissue ahead of the tumour volume during carbon
ion radiotherapy. Treatment with a single 4 Gy carbon ion dose resulted in 32% incidence of T-cell
lymphoma, double the incidence following the same dose of γ-rays. Across 100 carbon-ion induced
tumours, examination by LOH, gene sequencing and copy number analyses showed the most frequent
changes at Pten, Ikaros, Bcl11b and Notch1 loci. These genes are all also associated with X- and γ-ray-
induced T-cell lymphoma and were similarly activated/inactivated by a combination of point-mutations
or deletions, with or without loss of the remaining allele by mitotic recombination or copy number loss
depending on the locus. Although the lymphoma incidence was doubled, no novel carbon-ion specific
tumour suppressor gene targets were identified in these tumours. We are continuing to investigate
whether the chromosomal-level changes show evidence of increased complexity relative to photon-
induced tumours. Understanding the mechanisms of carbon-ion radiation-induced second tumours will
assist in risk-assessment for the safety of carbon-ion radiotherapy in children.
(PS5-48) Role of jnk inhibition in metastatic oral cavity squamous cell carcinoma. Sophia Bornstein, MD
PhD; John Gleysteen, MD; Casey Kernan, BS; David Sauer, MD; Charles R. Thomas, MD; and Melissa H.
Wong, PhD; OHSU, Portland, OR
Purpose/Objectives: Oral cavity squamous cell carcinoma (OSCC) is a common neoplasm
worldwide and is characterized by poor prognosis and low survival rate despite sophisticated surgical and
radiotherapeutic modalities. Lymph node spread significantly contributes to disease morbidity and
mortality in this population. The role of the noncanonical Wnt planar cell polarity pathway and
downstream Jnk signaling in lymph node metastases is unclear. Our objective was to evaluate the role of
Wnt5a and Jnk signaling in promoting lymph node metastatic OSCC in patient samples and in vitro.
Materials/Methods: We immunostained our in house oral cavity tissue microarrays using an antibody
against Wnt5a. We analyzed the effect of Wnt5a signaling on OSCC OSC19 and Cal27 cell lines alone and
in combination with non-canonical Jnk signaling inhibitor SP600125 or control canonical inhibitor DKK-1.
Downstream signaling assays were characterized using Western blot. Functional 3D invasion assays using
matrigel were quantitatively compared using IncuCYTE live imaging. Results: Wnt5a was overexpressed in
lymph node samples on the TMA compared to primary samples. Wnt5a led to increased Jnk signaling that
was blocked by Jnk inhibitor SP600125 but not canonical pathway inhibitor DKK-1. Wnt5a led to increased
matrigel invasion that was blocked by Jnk inhibition using SP600125 but not canonical pathway inhibition
312 | P a g e
induced second tumours. Benjamin J. Blyth, PhD; Shizuko Kakinuma; Yutaka Yamada; Takamitsu Morioka;
Shinobu Hirano-Sakairi; Yoshiko Amasaki; Yi Shang; Chizuru Tsuruoka; Tatsuhiko Imaoka; Mayumi
Nishimura; and Yoshiya Shimada, National Institute of Radiological Sciences, Chiba, Japan
The National Institute of Radiological Sciences, Japan, utilises a range of experimental animal
models in both short-term and lifespan studies to investigate tumours across an array of tissue sites
induced by X-ray, γ-ray, neutron or heavy-ion exposure, focusing on carbon ion radiotherapy as performed
at the HIMAC facility (Heavy Ion Medical Accelerator in Chiba) at our institute in particular. We are in the
process of establishing ‘J-Share’, an online archive to provide access to experimental data from our
collection of lifespan/cancer incidence studies, to ultimately facilitate the world-wide sharing of data and
archived tissues for collaborative endeavours. Here, we report on one such analysis of archived samples
of T-cell lymphomas induced by whole-body exposure(s) of 1-week-old mice to a monoenergetic carbon
ion beam (13 keV) simulating the exposure of normal tissue ahead of the tumour volume during carbon
ion radiotherapy. Treatment with a single 4 Gy carbon ion dose resulted in 32% incidence of T-cell
lymphoma, double the incidence following the same dose of γ-rays. Across 100 carbon-ion induced
tumours, examination by LOH, gene sequencing and copy number analyses showed the most frequent
changes at Pten, Ikaros, Bcl11b and Notch1 loci. These genes are all also associated with X- and γ-ray-
induced T-cell lymphoma and were similarly activated/inactivated by a combination of point-mutations
or deletions, with or without loss of the remaining allele by mitotic recombination or copy number loss
depending on the locus. Although the lymphoma incidence was doubled, no novel carbon-ion specific
tumour suppressor gene targets were identified in these tumours. We are continuing to investigate
whether the chromosomal-level changes show evidence of increased complexity relative to photon-
induced tumours. Understanding the mechanisms of carbon-ion radiation-induced second tumours will
assist in risk-assessment for the safety of carbon-ion radiotherapy in children.
(PS5-48) Role of jnk inhibition in metastatic oral cavity squamous cell carcinoma. Sophia Bornstein, MD
PhD; John Gleysteen, MD; Casey Kernan, BS; David Sauer, MD; Charles R. Thomas, MD; and Melissa H.
Wong, PhD; OHSU, Portland, OR
Purpose/Objectives: Oral cavity squamous cell carcinoma (OSCC) is a common neoplasm
worldwide and is characterized by poor prognosis and low survival rate despite sophisticated surgical and
radiotherapeutic modalities. Lymph node spread significantly contributes to disease morbidity and
mortality in this population. The role of the noncanonical Wnt planar cell polarity pathway and
downstream Jnk signaling in lymph node metastases is unclear. Our objective was to evaluate the role of
Wnt5a and Jnk signaling in promoting lymph node metastatic OSCC in patient samples and in vitro.
Materials/Methods: We immunostained our in house oral cavity tissue microarrays using an antibody
against Wnt5a. We analyzed the effect of Wnt5a signaling on OSCC OSC19 and Cal27 cell lines alone and
in combination with non-canonical Jnk signaling inhibitor SP600125 or control canonical inhibitor DKK-1.
Downstream signaling assays were characterized using Western blot. Functional 3D invasion assays using
matrigel were quantitatively compared using IncuCYTE live imaging. Results: Wnt5a was overexpressed in
lymph node samples on the TMA compared to primary samples. Wnt5a led to increased Jnk signaling that
was blocked by Jnk inhibitor SP600125 but not canonical pathway inhibitor DKK-1. Wnt5a led to increased
matrigel invasion that was blocked by Jnk inhibition using SP600125 but not canonical pathway inhibition
312 | P a g e
