Tomoaki Tamaki

Upregulation of stress-response genes with cell cycle arrest induced by carbon ion irradiation in multiple murine tumors models

Objective: To elucidate the in vivo biological effects induced by carbon-ion irradiation using comprehensive expression analysis. Materials and Methods: We examined gene expression changes after carbon-ion (C-ion) irradiation (290 MeV/m, SOBP 6 cm middle, 50 kev/μm) with a single dose of 30 Gy in four mouse tumors (NR-S1, SCCVII, NFSa, and #8520) transplanted into the hind legs of C3H/HeNrs mice, using 44K single-color oligo-microarrays at 6 hours (h), 1 day, and 3 days after irradiation. Gamma rays of 30 Gy and 50 Gy were used as a reference beam. Identification of C-ion-responsive genes was based on a false discovery rate of < 5% using the Wilcoxon test (P < 0.001) and the Benjamini-Hochberg correction. Results: In all tumors, the level of expression of several tens of genes, including Ccl3, Ccng1, Cd80, Cdkn1a, Cxcl2, IL7r, Lrdd, Mgmt, Mmp8, and Polk, was significantly altered 6 h and day 1 following C-ion irradiation. At day 3, several hundred genes, many of which are also classified as stress-response or cell-communication genes, including Tnfrsf5, Ikbke, and Icam1, were upregulated following C-ion irradiation. The expression level of the majority of these genes was similar following γ-ray treatment, although the change was not as extensive and intertumor variance was apparent. Several genes, including Ikbke, Serpina3n, and Saa3, responded differentially following C-ion irradiation than after γ-ray irradiation. Pathological investigation and immunohistochemical analysis of Cdkn1a revealed cell cycle arrest with mitotic catastrophe in tumors irradiated by C-ions. Conclusions: This study revealed significant C-ion induced upregulation of stress-responsive and cell-communication genes common to different tumor types. These findings provide evidence for the efficacy of this modality for the treatment of local tumors.

Radiotherapy plus Concomitant Adjuvant Temozolomide for Glioblastoma: Japanese Mono-Institutional Results

This study was conducted to investigate the feasibility and survival benefits of combined treatment with radiotherapy and temozolomide (TMZ), which has been covered by the national health insurance in Japanese patients with glioblastoma since September 2006. Between September 2006 and December 2011, 47 patients with newly diagnosed and histologically confirmed glioblastoma received radiotherapy for 60 Gy in 30 fractions. Among them, 45 patients (TMZ group) received concomitant TMZ (75 mg/m2/day, every day) and adjuvant TMZ (200 mg/m2/day, 5 days during each 28-days). All 36 of the glioblastoma patients receiving radiotherapy between January 1988 and August 2006 were analyzed as historical controls (control group). All patients were followed for at least 1 year or until they died. The median survival was 15.8 months in the TMZ group and 12.0 months in the control group after a median follow-up of 14.0 months. The hazard ratio for death in the TMZ group relative to the control group was 0.52 (P<0.01); the 2-year survival rate was 27.7% in the TMZ group and 14.6% in the control group. Hematologic toxicity of grade 3 and higher was observed in 20.4% in the TMZ group. Multivariate analysis showed that extent of surgery had the strongest impact on survival (P<0.01), while the use of TMZ had the second largest impact on survival (P = 0.035). The results indicate that combined treatment with radiotherapy and TMZ has a significant survival benefit for Japanese patients with newly diagnosed glioblastoma with slightly higher toxicities than previously reported.

Clinical outcomes of carbon ion radiotherapy for locally advanced adenocarcinoma of the uterine cervix in phase 1/2 clinical trial (protocol 9704).

This study sought to evaluate the toxicity and efficacy of carbon ion radiotherapy (C‐ion RT) for locally advanced adenocarcinoma of the uterine cervix in a phase 1/2 clinical trial.

Application of Carbon-Ion Beams or Gamma-Rays on Primary Tumors Does Not Change the Expression Profiles of Metastatic Tumors in an In Vivo Murine Model

PURPOSE To clarify how carbon-ion radiotherapy (C-ion) on primary tumors affects the characteristics of subsequently arising metastatic tumor cells. METHODS AND MATERIALS Mouse squamous cell carcinomas, NR-S1, in synergic C3H/HeMsNrs mice were irradiated with a single dose of 5-50 Gy of C-ion (290 MeV per nucleon, 6-cm spread-out Bragg peak) or gamma-rays ((137)Cs source) as a reference beam. The volume of the primary tumors and the number of metastatic nodules in lung were studied, and histologic analysis and microarray analysis of laser-microdissected tumor cells were also performed. RESULTS Including 5 Gy of C-ion and 8 Gy of gamma-rays, which did not inhibit the primary tumor growth, all doses used in this study inhibited lung metastasis significantly. Pathologic findings showed no difference among the metastatic tumor nodules in the nonirradiated, C-ion-irradiated, and gamma-ray-irradiated groups. Clustering analysis of expression profiles among metastatic tumors and primary tumors revealed a single cluster consisting of metastatic tumors different from their original primary tumors, indicating that the expression profiles of the metastatic tumor cells were not affected by the local application of C-ion or gamma-ray radiotherapy. CONCLUSION We found no difference in the incidence and histology, and only small differences in expression profile, of distant metastasis between local C-ion and gamma-ray radiotherapy. The application of local radiotherapy per se or the type of radiotherapy applied did not influence the transcriptional changes caused by metastasis in tumor cells.

Long-term results of curative intraluminal high dose rate brachytherapy for endobronchial carcinoma

BackgroundThe treatment strategy of central lung tumors is not established. Intraluminal brachytherapy (ILBT) is widely used for palliative treatment of endobronchial tumors, however, it is also a promising option for curative treatment with limited data. This study evaluates the results after ILBT for endobronchial carcinoma.MethodSixteen-endobronchial carcinoma of 13 patients treated with ILBT in curative intent for 2000 to 2008 were retrospectively reviewed. ILBT using high dose rate 192 iridium thin wire system was performed with 5 Gy/fraction at mucosal surface. The patient age ranged from 57 to 82 years old with median 75 years old. The 16 lesions consisted of 13 central endobronchial cancers including 7 roentgenographically occult lung cancers and 3 of tracheal cancers. Of them, 10 lesions were treated with ILBT of median 20 Gy combined with external beam radiation therapy of median 45 Gy and 6 lesions were treated with ILBT alone of median 25 Gy.ResultsMedian follow-up time was 32.5 months. Two-year survival rate and local control rate were 92.3% and 86.2%, respectively. Local recurrences were observed in 2 lesions. Three patients died due to lung cancer (1 patient) and intercurrent disease (2 patients). Complications greater than grade 2 were not observed except for one grade 3 dyspnea.ConclusionsILBT combined with or without EBRT might be a curative treatment option in inoperable endobronchial carcinoma patients with tolerable complication.

Prognostic value of FDG-PET in patients with ovarian carcinoma following surgical treatment

ObjectiveTo determine the prognostic value of FDG-PET after surgical resection in patients with ovarian carcinoma, we compared the results of FDG-PET and serum CA-125 level and prognosis of patients.Methods: Eighteen patients underwent a total of 32 FDG-PET examinations following surgery for ovarian carcinoma from October 2001 to December 2002 at our hospital (median follow-up period, 31 months). Age of the patients at the time of the initial FDG-PET examination ranged from 31 to 73 years (mean 52 years) and the period from surgery to the initial FDG-PET examination ranged from 5 to 109 months (mean 30 months). Serum CA-125 levels were determined on the occasion of each FDG-PET examination. Recurrent tumors were treated with surgery in 5 cases, radiotherapy in 2 cases, and chemotherapy in 9 cases.ResultsThe initial FDG-PET examinations revealed that 13 cases had positive and 5 cases had negative findings, which included 2 false positive cases. The survival rate for all patients at 1 year and 2 years after the initial examination was 82% and 63%, respectively. Two-year survival rates in patients with positive and negative FDG-PET findings were 51% and 83%, respectively, and the difference was not statistically significant (p = 0.19). Furthermore, 4 patients with normal CA-125 levels and 14 patients with elevated CA-125 levels showed 2-year survival rates of 100% and 51%, respectively, and they were not significantly different (p = 0.11). For all 32 examinations, the 2-year survival rates for patients with normal CA-125 levels (100%) were significantly higher (p = 0.025) than that for patients with elevated CA-125 levels (47%), however there was no significant difference (p = 0.20) between FDG-PET positive cases (53%) and negative cases (83%).ConclusionThe prognosis of patients with positive FDG-PET findings was less favorable than that of patients with negative findings. However, over the mean extended observation period of about 2.5 years, no significant difference in the prognosis of patients was observed between the two groups. The results of the present study indicate that elevated serum CA-125 levels may be more useful for evaluating the prognosis of ovarian cancer during the post-operative follow-up than FDG-PET findings.

Rectal bleeding after high-dose-rate brachytherapy combined with hypofractionated external-beam radiotherapy for localized prostate cancer: the relationship between dose-volume histogram parameters and the occurrence rate.

PURPOSE To determine the predictive risk factors for Grade 2 or worse rectal bleeding after high-dose-rate brachytherapy (HDR-BT) combined with hypofractionated external-beam radiotherapy (EBRT) for prostate cancer using dose-volume histogram analysis. METHODS AND MATERIALS The records of 216 patients treated with HDR-BT combined with EBRT were analyzed. The treatment protocols for HDR-BT were 5 Gy × five times in 3 days or 7 Gy × three, 10.5 Gy × two, or 9 Gy × two in 2 days. The EBRT doses ranged from 45 to 51 Gy with a fractional dose of 3 Gy. RESULTS In 20 patients Grade 2 or worse rectal bleeding developed, and the cumulative incidence rate was 9% at 5 years. By converting the HDR-BT and EBRT radiation doses into biologic effective doses (BED), the BED(3) at rectal volumes of 5% and 10% in the patients who experienced bleeding were significantly higher than those in the remaining 196 patients. Univariate analysis showed that a higher rectal BED(3-5%) and the use of fewer needles in brachytherapy were correlated with the incidence of bleeding, but BED(3-5%) was found to be the only significant factor on multivariate analysis. CONCLUSIONS The radiation dose delivered to small rectal lesions as 5% is important for predicting Grade 2 or worse rectal bleeding after HDR-BT combined with EBRT for prostate cancer.

Filling the gap in central shielding: three-dimensional analysis of the EQD2 dose in radiotherapy for cervical cancer with the central shielding technique

This study aimed to provide accurate dose distribution profiles of radiotherapy for cervical cancer when treated with the central shielding technique by analysing the composite 3D EQD2 dose distribution of external beam radiotherapy (EBRT) plus intracavitary brachytherapy (ICBT). On a phantom, four patterns of the combinations of whole pelvis irradiation (WP) (4 fields), pelvis irradiation with central shielding technique (CS) [anterior–posterior/posterior–anterior (AP-PA fields), shielding width of 3 or 4 cm] and ICBT using Point-A prescription were created: 30 Gy/15 fractions + 20 Gy/10 fractions + 24 Gy/4 fractions [Plan (30 + 20 + 24)], 40 Gy/20 fractions + 10 Gy/5 fractions + 18 Gy/3 fractions [Plan (40 + 10 + 18)], 40 Gy/20 fractions + 10 Gy/5 fractions + 24 Gy/4 fractions [Plan (40 + 10 + 24)] and 45 Gy/25 fractions + 0 Gy + 28 Gy/4 fractions [Plan (45 + 0 + 28)]. The composite EQD2 dose distributions of the complete treatment were analysed. The Point-A dose of Plan (30 + 20 + 24), Plan (40 + 10 + 18), Plan (40 + 10 + 24) and Plan (45 + 0 + 28) were 78.0 Gy (CS 3 cm)/71.8 Gy (CS 4 cm), 72.1 Gy (CS 3 cm)/69.0 Gy (CS 4 cm), 80.1 Gy (CS 3 cm)/77.0 Gy (CS 4 cm) and 84.1 Gy, whereas it has been previously reported to be 62 Gy, 64 Gy, 72 Gy and 84 Gy, respectively. For all the treatment plans with CS, equivalent or wider coverage of 60 Gy (EQD2) was achieved in the right–left direction, while coverage in the anterior–posterior direction decreased in plans with CS. There were no irregularly ‘cold’ regions around the central target. The use of CS in radiotherapy for cervical cancer resulted in tumor coverage in the lateral direction with doses higher than the previously reported Point-A doses.

Comparison of hematological toxicities between innovator and generic cisplatin formulations in cervical cancer patients treated with concurrent chemoradiotherapy

To compare the incidence and degree of hematological toxicity between innovator and generic cisplatin formulations, decreases in white blood cell (WBC) count (leukopenia) and platelet counts (thrombocytopenia) were retrospectively examined, using the Common Toxicity Criteria for Adverse Events ver. 4.0, in patients with uterine cervical cancer treated with concurrent chemoradiotherapy using innovator (innovator group, n = 22) or generic (generic group, n = 22) cisplatin formulations. There were no significant differences in patient characteristics except in the technique of external irradiation; larger numbers of patients in the innovator and generic groups were irradiated using the parallel-opposed two-field technique and the four-field box technique, respectively (P = 0.00012), which is in line with the historical progress of external beam radiation therapy. The numbers of patients showing Grade 1, 2, 3 and 4 leukopenia were 1 (4.5%), 14 (64%), 7 (32%) and 0 (0.0%) in the innovator group, and 1 (4.5%), 6 (27%), 13 (59%) and 2 (9.0%) in the generic group, respectively. The number of patients showing Grade 3–4 leukopenia was significantly greater in the generic group than in the innovator group (P = 0.034). There was no significant relationship between the incidence of Grade 3–4 leukopenia and the technique of external irradiation. There were no significant differences in the incidence and degree of thrombocytopenia between the two groups. These results indicate the possibility that the generic cisplatin formulation may have a different toxicity profile compared to the innovator formulation in terms of the incidence of leukopenia.

Villin1, a novel diagnostic marker for cervical adenocarcinoma

The number of new cervical adenocarcinoma (AD) cases has risen slowly, however, its histological similarity to other tumor types and the difficulty of identifying the site of the original tumor makes the diagnosis of cervical AD particularly challenging. We investigated a novel molecular biomarker for cervical AD through the integration of multiple methods of genomic analysis. Tumor samples in discovery set were obtained from 87 patients who underwent radiotherapy, including 31 cervical AD. Microarray analysis and quantitative polymerase chain reaction analysis were performed to screen a candidate diagnostic molecule for cervical AD, and its clinical significance was investigated by immunohistochemical analysis (IHC). We found a difference between biopsy samples of AD and squamous cell carcinoma (SCC) in the expression and genomic copy number of Villin1 (VIL1), which maps to 2q35. IHC revealed 14 VIL1-positive tumors; 13 cervical AD and 1 small cell carcinoma of cervix, while none of SCC or endometrial AD was VIL1-positive. Kaplan-Meier survival curves revealed worse disease-free survival in VIL1-positive tumors. The marker was validated by newly enrolled 65 patients, and VIL1 positive staining showed 52% of sensitivity and 100% of selectivity for cervical AD. In conclusion, we have identified VIL1 as a novel biomarker of cervical AD. VIL1, a major structural component of the brush border cytoskeleton, which was recently found to be an epithelial cell-specific anti-apoptotic protein. Our study suggests the existence of a subtype of cervical tumors which are VIL1 positive with poor radioresponse.