Joseph T. Rakowski

Soy isoflavones radiosensitize lung cancer while mitigating normal tissue injury

BACKGROUND We have demonstrated that soy isoflavones radiosensitize cancer cells. Prostate cancer patients receiving radiotherapy (RT) and soy tablets had reduced radiation toxicity to surrounding organs. We have now investigated the combination of soy with RT in lung cancer (NSCLC), for which RT is limited by radiation-induced pneumonitis. METHODS Human A549 NSCLC cells were injected i.v. in nude mice to generate lung tumor nodules. Lung tumor-bearing mice were treated with left lung RT at 12 Gy and with oral soy treatments at 1mg/day for 30 days. Lung tissues were processed for histology. RESULTS Compared to lung tumor nodules treated with soy isoflavones or radiation, lung tissues from mice treated with both modalities showed that soy isoflavones augmented radiation-induced destruction of A549 lung tumor nodules leading to small residual tumor nodules containing degenerating tumor cells with large vacuoles. Soy isoflavones decreased the hemorrhages, inflammation and fibrosis caused by radiation in lung tissue, suggesting protection of normal lung tissue. CONCLUSIONS Soy isoflavones augment destruction of A549 lung tumor nodules by radiation, and also mitigate vascular damage, inflammation and fibrosis caused by radiation injury to normal lung tissue. Soy could be used as a non-toxic complementary approach to improve RT in NSCLC.

Differential effect of soy isoflavones in enhancing high intensity radiotherapy and protecting lung tissue in a pre-clinical model of lung carcinoma

BACKGROUND Radiotherapy of locally-advanced non-small cell lung cancer is limited by radiation-induced pneumonitis and fibrosis. We have further investigated the role of soy isoflavones to improve the effect of a high intensity radiation and reduce lung damage in a pre-clinical lung tumor model. METHODS Human A549 NSCLC cells were injected i.v. in nude mice to generate a large tumor burden in the lungs. Mice were treated with lung irradiation at 10 Gy and with oral soy. The therapy effect on the tumor cells and surrounding lung tissue was analyzed on lung sections stained with H&E, Ki-67 and Massons Trichrome. Pneumonitis and vascular damage were evaluated by measurements of alveolar septa and immunofluorescent staining of vessel walls. RESULTS Combined soy and radiation caused a significantly stronger inhibition of tumor progression compared to each modality alone in contrast to large invasive tumor nodules seen in control mice. At the same time, soy reduced radiation injury in lung tissue by decreasing pneumonitis, fibrosis and protecting alveolar septa, bronchioles and vessels. CONCLUSIONS These studies demonstrate a differential effect of soy isoflavones on augmenting tumor destruction induced by radiation while radioprotecting the normal lung tissue and support using soy to alleviate radiotoxicity in lung cancer.

Radioprotection of Lung Tissue by Soy Isoflavones

Introduction: Radiation-induced pneumonitis and fibrosis have restricted radiotherapy for lung cancer. In a preclinical lung tumor model, soy isoflavones showed the potential to enhance radiation damage in tumor nodules and simultaneously protect normal lung from radiation injury. We have further dissected the role of soy isoflavones in the radioprotection of lung tissue. Methods: Naive Balb/c mice were treated with oral soy isoflavones for 3 days before and up to 4 months after radiation. Radiation was administered to the left lung at 12 Gy. Mice were monitored for toxicity and breathing rates at 2, 3, and 4 months after radiation. Lung tissues were processed for histology for in situ evaluation of response. Results: Radiation caused damage to normal hair follicles, leading to hair loss in the irradiated left thoracic area. Supplementation with soy isoflavones protected mice against radiation-induced skin injury and hair loss. Lung irradiation also caused an increase in mouse breathing rate that was more pronounced by 4 months after radiation, probably because of the late effects of radiation-induced injury to normal lung tissue. However, this effect was mitigated by soy isoflavones. Histological examination of irradiated lungs revealed a chronic inflammatory infiltration involving alveoli and bronchioles and a progressive increase in fibrosis. These adverse effects of radiation were alleviated by soy isoflavones. Conclusion: Soy isoflavones given pre- and postradiation protected the lungs against adverse effects of radiation including skin injury, hair loss, increased breathing rates, inflammation, pneumonitis and fibrosis, providing evidence for a radioprotective effect of soy.

Gamma Knife radiosurgery in the management of brainstem metastases.

BACKGROUND Metastases to the brainstem portend a poor prognosis and present a challenge in clinical management. Surgical resection is rarely a viable option. METHODS Post-treatment MRI scans of patients with brainstem metastases treated with radiosurgery were used to determine local control and disease progression. Median survival was calculated using Kaplan-Meier analysis. Univariate and multivariate analyses were performed using log-rank test and Cox proportional hazards model, respectively. RESULTS Thirty-two consecutive patients with brainstem metastasis underwent Gamma Knife radiosurgery. Median age was 50 years. Median tumor volume was 0.71 cm3 and median tumor margin dose was 13 Gy. Seventeen of 32 patients received WBRT prior to stereotactic radiosurgery. Median survival was 5.2 months. There was a statistically significant difference in survival based on RTOG recursive partition analysis (RPA) class. Median survival of patients categorized as RPA class I was 19.2 months, RPA class II was 8.4 months, and RPA class III was 1.9 months. The overall local tumor control rate was 87.5%. There were no acute complications following stereotactic radiosurgery and no evidence of radiation necrosis noted on post-treatment MRI scans. CONCLUSION Stereotactic radiosurgery is an effective treatment for brainstem metastases and should be considered especially for patients with good performance status.

Soy Isoflavones Promote Radioprotection of Normal Lung Tissue by Inhibition of Radiation-Induced Activation of Macrophages and Neutrophils

Introduction: Radiation therapy for lung cancer is limited by toxicity to normal lung tissue that results from an inflammatory process, leading to pneumonitis and fibrosis. Soy isoflavones mitigate inflammatory infiltrates and radiation-induced lung injury, but the cellular immune mediators involved in the radioprotective effect are unknown. Methods: Mice received a single dose of 10 Gy radiation delivered to the lungs and daily oral treatment of soy isoflavones. At different time points, mice were either processed to harvest bronchoalveolar lavage fluid for differential cell counting and lungs for flow cytometry or immunohistochemistry studies. Results: Combined soy and radiation led to a reduction in infiltration and activation of alveolar macrophages and neutrophils in both the bronchoalveolar and lung parenchyma compartments. Soy treatment protected F4/80+CD11c− interstitial macrophages, which are known to play an immunoregulatory role and are decreased by radiation. Furthermore, soy isoflavones reduced the levels of nitric oxide synthase 2 expression while increasing arginase-1 expression after radiation, suggesting a switch from proinflammatory M1 macrophage to an anti-inflammatory M2 macrophage phenotype. Soy also prevented the influx of activated neutrophils in lung caused by radiation. Conclusions: Soy isoflavones inhibit the infiltration and activation of macrophages and neutrophils induced by radiation in lungs. Soy isoflavones-mediated modulation of macrophage and neutrophil responses to radiation may contribute to a mechanism of resolution of radiation-induced chronic inflammation leading to radioprotection of lung tissue.

Characteristic 8 keV X Rays Possess Radiobiological Properties of Higher-LET Radiation

Abstract Electronic brachytherapy systems are being developed that can deliver X rays of varying energy depending on the material of a secondary target. A copper target produces characteristic 8 keV X rays. Our aim was to determine whether 8 keV X rays might deliver greater biological effectiveness than megavoltage photons. Cells of the U251 human glioma cell line were used to compare the biological effects of 8 keV X rays and 60Co γ rays in terms of relative biological effectiveness (RBE), oxygen enhancement ratio (OER), and DNA damage. The RBE at 50% and 10% survival was 2.6 and 1.9, respectively. At 50% survival, the OER for cells treated with 8 keV X rays was 1.6 compared with 3.0 for 60Co γ rays. The numbers of H2AX foci per Gy after treatment with 8 keV X rays and 60Co γ rays were similar; however, the size of the foci generated at 8 keV was significantly larger, possibly indicating more complex DNA damage. The mean area of H2AX foci generated by 8 keV X rays was 0.785 µm2 (95% CI: 0.756–0.814) compared with 0.491 µm2 (95% CI: 0.462–0.520) for 60Co γ rays (P < 0.0001). Characteristic 8 keV X rays produce two to three times the biological effectiveness of megavoltage photons, with a radiobiological profile similar to higher-LET radiations.

Helical TomoTherapy versus sterotactic Gamma Knife radiosurgery in the treatment of single and multiple brain tumors: a dosimetric comparison

The objective was to compare the dosimetry of Helical TomoTherapy (TOMO) and Gamma Knife (GK) treatment plans for tumor and normal brain in the treatment of single and multiple brain tumors. An anthropomorphic Rando Head phantom was used to compare the dosimetry of TOMO and GK. Eight brain tumors of various shapes, sizes and locations were used to generate 10 plans. The radiation dose was 20 Gy prescribed to the 100% isodose line for TOMO plans and to the 50% for the GK plans. Dose Volume Histograms for tumor and brain were compared. Equivalent Uniform Dose (gEUD), Tumor Control Probability (TCP) and Normal Tissue Complication Probability (NTCP) were performed and used for plan comparisons. Average minimum, mean, median and maximum tumor doses were 19.93, 27.83, 27.38, 39.60 Gy for GK and 20.17, 20.60, 20.59, 20.90 Gy for TOMO. Average gEUD values for tumor and normal brain were 25.0 and 7.2 Gy for GK and 20.7 and 8.1 Gy for TOMO. Conformity indices (CI) were similar for both modalities. Gradient indices (GI) were greater for TOMO. A combination plan was also generated using all eight tumors. TOMO was able to target all eight tumors simultaneously resulting in mean tumor and brain doses of 20.5 and 9.35 Gy, respectively. Due to the maximum limit of 50 beams per plan, GK was unable to provide a treatment plan for all eight tumors. GK provides an advantage for all tumor sizes with respect to tumor and normal brain dose. Clinical studies are needed to correlate these dosimetric findings with patient outcomes. PACS number: 87.55‐x

Approaches to interventional fluoroscopic dose curves

Modern fluoroscopes used for image‐based guidance in interventional procedures are complex X‐ray machines, with advanced image acquisition and processing systems capable of automatically controlling numerous parameters based on defined protocol settings. This study evaluated and compared approaches to technique factor modulation and air kerma rates in response to simulated patient thickness variations for four state‐of‐the‐art and one previous‐generation interventional fluoroscopes. A polymethyl methacrylate (PMMA) phantom was used as a tissue surrogate for the purposes of determining fluoroscopic reference plane air kerma rates, kVp, mA, and variable copper filter thickness over a wide range of simulated tissue thicknesses. Data were acquired for each fluoroscopic and acquisition dose curve within each vendors default abdomen or body imaging protocol. The data obtained indicated vendor‐ and model‐specific variations in the approach to technique factor modulation and reference plane air kerma rates across a range of tissue thicknesses. However, in the imaging protocol evaluated, all of the state‐of‐the‐art systems had relatively low air kerma rates in the fluoroscopic low‐dose imaging mode as compared to the previous‐generation unit. Each of the newest‐generation systems also employ Cu filtration within the selected protocol in the acquisition mode of imaging; this is a substantial benefit, reducing the skin entrance dose to the patient in the highest dose‐rate mode of fluoroscope operation. Some vendors have also enhanced the radiation output capabilities of their fluoroscopes which, under specific conditions, may be beneficial; however, these increased output capabilities also have the potential to lead to unnecessarily high dose rates. Understanding how fluoroscopic technique factors are modulated provides insight into the vendor‐specific image acquisition approach and may provide opportunities to optimize the imaging protocols for clinical practice. PACS number: 87.59.C‐Modern fluoroscopes used for image-based guidance in interventional procedures are complex X-ray machines, with advanced image acquisition and processing systems capable of automatically controlling numerous parameters based on defined protocol settings. This study evaluated and compared approaches to technique factor modulation and air kerma rates in response to simulated patient thickness variations for four state-of-the-art and one previous-generation interventional fluoroscopes. A polymethyl methacrylate (PMMA) phantom was used as a tissue surrogate for the purposes of determining fluoroscopic reference plane air kerma rates, kVp, mA, and variable copper filter thickness over a wide range of simulated tissue thicknesses. Data were acquired for each fluoroscopic and acquisition dose curve within each vendors default abdomen or body imaging protocol. The data obtained indicated vendor- and model-specific variations in the approach to technique factor modulation and reference plane air kerma rates across a range of tissue thicknesses. However, in the imaging protocol evaluated, all of the state-of-the-art systems had relatively low air kerma rates in the fluoroscopic low-dose imaging mode as compared to the previous-generation unit. Each of the newest-generation systems also employ Cu filtration within the selected protocol in the acquisition mode of imaging; this is a substantial benefit, reducing the skin entrance dose to the patient in the highest dose-rate mode of fluoroscope operation. Some vendors have also enhanced the radiation output capabilities of their fluoroscopes which, under specific conditions, may be beneficial; however, these increased output capabilities also have the potential to lead to unnecessarily high dose rates. Understanding how fluoroscopic technique factors are modulated provides insight into the vendor-specific image acquisition approach and may provide opportunities to optimize the imaging protocols for clinical practice. PACS number: 87.59.C.

Effects of titanium mesh implant on dosimetry during Gamma Knife radiosurgery

Calvarial reconstruction following resection of tumors involving the skull is often followed by stereotactic radiosurgery. Prior studies have addressed the effects of various cranioplasty materials on dose distributions in linac‐based radiosurgery. We aim to determine the effects of titanium mesh implants on Gamma Knife dose. Radiation backscatter and transmission were measured for eight types of titanium mesh using film, ion chamber, and Theratron Co‐60 teletherapy device. A single mesh was selected for Gamma Knife irradiation using a CaSO4 skull filled with ballistics gel. Dose profiles for reconstructed and intact skulls were compared with the planning system prediction at 2.5 and 5.5 cm depth. Titanium contact backscatter and transmission dose perturbations ranged from ‐18% to 23%. Radiation dose measured at 1.5 cm below the calvarial implant increased by 0.5% to 3.3% relative to bone. Measured Gamma Knife dose profile diameters agreed with expected profiles. Maximum dose within the intact phantom was 3% less than planned due to skull attenuation. Maximum dose within the reconstructed phantom was between the intact phantom and planned doses. Titanium mesh implants and hydroxyapatite cranioplasty result in minimal alteration (<3%) in the delivered Gamma Knife dose. PACS number: 87.00

Axitinib Improves Radiotherapy in Murine Xenograft Lung Tumors

A third of patients with non-small cell lung cancer (NSCLC) present with un-resectable stage III locally advanced disease and are currently treated by chemo-radiotherapy but the median survival is only about 21 months. Using an orthotopic xenograft model of lung carcinoma, we have investigated the combination of radiotherapy with the anti-angiogenic drug axitinib (AG-013736, Pfizer), which is a small molecule receptor tyrosine kinase inhibitor that selectively targets the signal transduction induced by VEGF binding to VEGFR receptors. We have tested the combination of axitinib with radiotherapy in nude mice bearing human NSCLC A549 lung tumors. The therapy effect was quantitatively evaluated in lung tumor nodules. The modulation of radiation-induced pneumonitis, vascular damage and fibrosis by axitinib was assessed in lung tissue. Lung irradiation combined with long-term axitinib treatment was safe resulting in minimal weight loss and no vascular injury in heart, liver and kidney tissues. A significant decrease in the size of lung tumor nodules was observed with either axitinib or radiation, associated with a decrease in Ki-67 staining and a heavy infiltration of inflammatory cells in tumor nodules. The lungs of mice treated with radiation and axitinib showed a complete response with no detectable residual tumor nodules. A decrease in pneumonitis, vascular damage and fibrosis were observed in lung tissues from mice treated with radiation and axitinib. Our studies suggest that axitinib is a potent and safe drug to use in conjunction with radiotherapy for lung cancer that could also act as a radioprotector for lung tissue by reducing pneumonitis and fibrosis.