Hiroaki Kunogi

Prediction of radiosensitivity using phosphorylation of histone H2AX and apoptosis in human tumor cell lines.

Abstract Purpose: We examined the relationship between radiosensitivity, histone H2AX (γH2AX) phosphorylation, and apoptosis to develop a new predictive assay for radiosensitivity. Materials and methods: Seven human tumor cell lines, including one fibrosarcoma (HT1080), four oesophageal carcinomas (TE-9, KYSE30, KYSE150, and KYSE220), and two breast carcinomas (HCC70, and ZR75-1) were used. Cellular radiosensitivity was assessed using a standard colony-forming assay. To measure the frequency of γH2AX foci, we counted the number of foci per cell under fluorescence microscopy following immunofluorescence staining. DNA content was determined by a flow cytometric assay. To assess the frequency of apoptosis, we enumerated apoptotic cells by fluorescence microscopy 24 hours after irradiation. Results: All seven cell lines showed dose (0–9 Gy)-dependent increases in the number of γH2AX foci per cell 24 h after irradiation. When both the frequency of γH2AX foci normalized by DNA content and the frequency of apoptosis were used, a better correlation was observed between the actual cell survivals and the predicted ones. Conclusions: Our study shows that the number of γH2AX foci after normalization of the DNA content and apoptotic cell frequency can be used as a new predictive assay for cell survival.

A new two-step accurate CT-MRI fusion technique for post-implant prostate cancer.

Purpose To develop an accurate method of fusing computed tomography (CT) with magnetic resonance imaging (MRI) for post-implant dosimetry after prostate seed implant brachytherapy. Material and methods Prostate cancer patients were scheduled to undergo CT and MRI after brachytherapy. We obtained the three MRI sequences on fat-suppressed T1-weighted imaging (FST1-WI), T2-weighted imaging (T2-WI), and T2*-weighted imaging (T2*-WI) in each patient. We compared the lengths and widths of 450 seed source images in the 10 study patients on CT, FST1-WI, T2-WI, and T2*-WI. After CT-MRI fusion using source positions by the least-squares method, we decided the center of each seed source and measured the distance of these centers between CT and MRI to estimate the fusion accuracy. Results The measured length and width of the seeds were 6.1 ± 0.5 mm (mean ± standard deviation) and 3.2 ± 0.2 mm on CT, 5.9 ± 0.4 mm, and 2.4 ± 0.2 mm on FST1-WI, 5.5 ± 0.5 mm and 1.8 ± 0.2 mm on T2-WI, and 7.8 ± 1.0 mm and 4.1 ± 0.7 mm on T2*-WI, respectively. The measured source location shifts on CT/FST1-WI and CT/T2-WI after image fusion in the 10 study patients were 0.9 ± 0.4 mm and 1.4 ± 0.2 mm, respectively. The shift on CT/FST1-WI was less than on CT/T2-WI (p = 0.005). Conclusions For post-implant dosimetry after prostate seed implant brachytherapy, more accurate fusion of CT and T2-WI is achieved if CT and FST1-WI are fused first using the least-squares method and the center position of each source, followed by fusion of the FST1-WI and T2-WI images. This method is more accurate than direct image fusion.

Focal partial salvage low-dose-rate brachytherapy for local recurrent prostate cancer after permanent prostate brachytherapy with a review of the literature

Purpose To investigate the treatment results for focal partial salvage re-implantation against local recurrence after permanent prostate brachytherapy. Material and methods Between January 2010 and September 2015, 12 patients were treated with focal partial salvage re-implantation for local recurrence after low-dose-rate brachytherapy using 125I seeds. The focal clinical target volume (F-CTV) was delineated on positive biopsy areas in a mapping biopsy, combining the cold spots on the post-implant dosimetry for initial brachytherapy. The F-CTV was expanded by 3 mm to create the planning target volume (PTV) as a margin to compensate for uncertainties in image registration and treatment delivery. The prescribed dose to the PTV was 145 Gy. The characteristics and biochemical disease-free survival (BdFS) rates were analyzed. Genitourinary (GU) and gastrointestinal (GI) toxicities were evaluated using the Common Terminology Criteria for Adverse Events version 4. Results The median prostate-specific antigen (PSA) level at re-implantation was 4.09 ng/ml (range: 2.91-8.24 ng/ml). The median follow-up time was 56 months (range: 6-74 months). The median RD2cc and UD10 were 63 Gy and 159 Gy, respectively. The 4-year BdFS rate was 78%, which included non-responders. Biochemical recurrence occurred in two patients after 7 and 31 months, respectively. The former was treated with hormonal therapy after biochemical failure, and the latter underwent watchful waiting (PSA at the last follow-up of 53 months: 7.3 ng/ml) at the patients request. No patients had grade 3 GU/GI toxicities or died after salvage re-implantation. Conclusions The partial salvage low-dose-rate brachytherapy used to treat local recurrence after permanent prostate brachytherapy is well-tolerated, with high biochemical response rates. This treatment can be not only a method to delay chemical castration but also a curative treatment option in cases of local recurrence of prostate carcinoma after seed implantation.

Kidney-Sparing Methods for Extended-Field Intensity-Modulated Radiotherapy (EF-IMRT) in Cervical Carcinoma Treatment

Coplanar extended-field intensity-modulated radiation therapy (EF-IMRT) targeting the whole-pelvic and para-aortic lymph nodes in patients with advanced cervical cancer results in impaired creatinine clearance. An improvement in renal function cannot be expected unless low-dose (approximately 10 Gy) kidney exposure is reduced. The dosimetric method should be considered during EF-IMRT planning to further reduce low-dose exposure to the kidneys. To assess the usefulness of non-coplanar EF-IMRT with kidney-avoiding beams to spare the kidneys during cervical carcinoma treatment in dosimetric analysis between non-coplanar and coplanar EF-IMRT, we compared the doses of the target organ and organs at risk, including the kidney, in 10 consecutive patients. To estimate the influence of EFRT on renal dysfunction, creatinine clearance values after treatment were also examined in 18 consecutive patients. Of these 18 patients, 10 patients who were included in the dosimetric analysis underwent extended field radiation therapy (EFRT) with concurrent chemotherapy, and eight patients underwent whole-pelvis radiation therapy with concurrent chemotherapy to treat cervical carcinoma between April 2012 and March 2015 at our institution. In the dosimetric analysis, non-coplanar EF-IMRT was effective at reducing low-dose (approximately 10 Gy) exposure to the kidneys, thus maintaining target coverage and sparing other organs at risk, such as the small bowel, rectum, and bladder, compared with coplanar EF-IMRT. Renal function in all 10 patients who underwent EFRT, including coplanar EF-IMRT (with kidney irradiation), was low after treatment, and differed significantly from that of the eight patients who underwent WPRT (no kidney irradiation) 6 months after the first day of treatment (P = 0.005). In conclusion, non-coplanar EF-IMRT should be considered in patients with advanced cervical cancer, particularly in patients with a long life expectancy or with pre-existing renal dysfunction.

Effect of a urinary catheter on seed position and rectal and bladder doses in CT-based post-implant dosimetry for prostate cancer brachytherapy

Purpose To assess the variability in rectal and bladder dosimetric parameters determined according to post-implant computed tomography (CT) images in patients with or without a urethral catheter. Material and methods Patients with prostate cancer who were scheduled to undergo CT after brachytherapy between October 2012 and January 2014 were included. We obtained CT series with and without a urinary catheter in each patient. We compared the rectal and bladder doses in 18 patients on each CT series. Results The shifts in the seed positions between with and without a catheter in place were 1.3 ± 0.3 mm (mean ± standard deviation). The radiation doses to the rectum, as determined on the CT series, with a urethral catheter were higher than those on CT without a catheter (p < 0.001). Radiation doses to the bladder with a catheter were significantly lower than those without a catheter (p = 0.027). Conclusions Post-implant dosimetry (PID) with no catheter showed significantly lower rectal doses and higher bladder doses than those of PID with a catheter. We recommend the PID procedure for CT images in patients without a catheter. Use of CT with a catheter is limited to identifying urethral position.