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Purpose@#Exogenous epidermal growth factor (EGF) causes apoptosis in EGF receptor (EGFR)–overexpressing cell lines. The apoptosis-inducing factors could be a therapeutic target. We aimed to determine the mechanism of EGF-induced apoptosis using a genome-wide clustered regularly interspaced short palindromic repeats (CRISPR)-based knockout screen. @*Materials and Methods@#Two-vector system of the human genome-scale CRISPR knockout library v2 was used to target 19,050 genes using 123,411 single guide RNAs (sgRNAs). Recombinant human EGF (100 nM) or distilled water four times was administered to the experimental and control groups, respectively. The read counts of each sgRNA obtained from next-generation sequencing were analyzed using the edgeR algorithm. We used another EGFR-overexpressing cell line (A549) and short hairpin RNAs (shRNAs) targeting five EGF-resistance genes for validation. DUSP1 expression in A431, A549, and HEK293FT cells was calculated using reverse transcription–quantitative polymerase chain reaction. @*Results@#We found 77 enriched and 189 depleted genes in the experimental group using the CRISPR-based knockout screen and identified the top five EGF-resistance genes: DDX20, LHFP, REPS1, DUSP1, and KRTAP10-12. Transfecting shRNAs targeting these genes into A549 cells significantly increased the surviving fractions after EGF treatment, compared with those observed in the control shRNA-transfected cells. The expression ratio of DUSP1 (inhibits ERK signaling) increased in A431 and A549 cells after EGF treatment. However, DUSP1 expression remained unchanged in HEK293FT cells after EGF treatment. @*Conclusion@#The CRISPR-based knockout screen revealed 266 genes possibly responsible for EGF-induced apoptosis. DUSP1 might be a critical component of EGF-induced apoptosis and a novel target for EGFR-overexpressing cancers.
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Purpose@#This study aimed to evaluate the role of postoperative radiotherapy (PORT) in intracranial solitary fibrous tumor/hemangiopericytoma (SFT/HPC). @*Materials and Methods@#A total of 133 patients with histologically confirmed HPC were included from eight institutions. Gross total resection (GTR) and subtotal resection (STR) were performed in 86 and 47 patients, respectively. PORT was performed in 85 patients (64%). The prognostic effects of sex, age, performance, World Health Organization (WHO) grade, location, size, Ki-67, surgical extent, and PORT on local control (LC), distant metastasis-free survival (DMFS), progression-free survival (PFS), and overall survival (OS) were estimated by univariate and multivariate analyses. @*Results@#The 10-year PFS, and OS rates were 45%, and 71%, respectively. The multivariate analysis suggested that PORT significantly improved LC (p < 0.001) and PFS (p < 0.001). The PFS benefit of PORT was maintained in the subgroup of GTR (p=0.001), WHO grade II (p=0.001), or STR (p < 0.001). In the favorable subgroup of GTR and WHO grade II, PORT was also significantly related to better PFS (p=0.028). WHO grade III was significantly associated with poor DMFS (p=0.029). In the PORT subgroup, the 0-0.5 cm margin of the target volume showed an inferior LC to a large margin with 1.0-2.0 cm (p=0.021). Time-dependent Cox proportion analysis showed that distant failures were significantly associated with poor OS (p=0.003). @*Conclusion@#This multicenter study supports the role of PORT in disease control of intracranial SFT/HPC, irrespective of the surgical extent and grade. For LC, PORT should enclose the tumor bed with sufficient margin.
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Purpose@#The value of the genomic profiling by targeted gene-sequencing on radiation therapy response prediction was evaluated through integrated analysis including clinical information. Radiation response prediction model was constructed based on the analyzed findings. @*Materials and Methods@#Patients who had the tumor sequenced using institutional cancer panel after informed consent and received radiotherapy for the measurable disease served as the target cohort. Patients with irradiated tumor locally controlled for more than 6 months after radiotherapy were defined as the durable local control (DLC) group, otherwise, non-durable local control (NDLC) group. Significant genomic factors and domain knowledge were used to develop the Bayesian Network model to predict radiotherapy response. @*Results@#Altogether, 88 patients were collected for analysis. Of those, 41 (43.6%) and 47 (54.4%) patients were classified as the NDLC and DLC group, respectively. Somatic mutations of NOTCH2 and BCL were enriched in the NDLC group, whereas, mutations of CHEK2, MSH2, and NOTCH1 were more frequently found in the DLC group. Altered DNA repair pathway was associated with better local failure–free survival (hazard ratio, 0.40; 95% confidence interval, 0.19 to 0.86; p=0.014). Smoking somatic signature was found more frequently in the DLC group. Area under the receiver operating characteristic curve of the Bayesian network model predicting probability of 6-month local control was 0.83. @*Conclusion@#Durable radiation response was associated with alterations of DNA repair pathway and smoking somatic signature. Bayesian network model could provide helpful insights for high precision radiotherapy. However, these findings should be verified in prospective cohort for further individualization.
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PURPOSE: To investigate positional uncertainty and its correlation with clinical parameters in spine stereotactic body radiotherapy (SBRT) using thermoplastic mask (TM) immobilization. MATERIALS AND METHODS: A total of 21 patients who underwent spine SBRT for cervical or upper thoracic spinal lesions were retrospectively analyzed. All patients were treated with image guidance using cone beam computed tomography (CBCT) and 4 degrees-of-freedom (DoF) positional correction. Initial, pre-treatment, and post-treatment CBCTs were analyzed. Setup error (SE), pre-treatment residual error (preRE), post-treatment residual error (postRE), intrafraction motion before treatment (IM1), and intrafraction motion during treatment (IM2) were determined from 6 DoF manual rigid registration. RESULTS: The three-dimensional (3D) magnitudes of translational uncertainties (mean ± 2 standard deviation) were 3.7±3.5 mm (SE), 0.9±0.9 mm (preRE), 1.2±1.5 mm (postRE), 1.4±2.4 mm (IM1), and 0.9±1.0 mm (IM2), and average angular differences were 1.1°±1.2° (SE), 0.9°±1.1° (preRE), 0.9°±1.1° (postRE), 0.6°±0.9° (IM1), and 0.5°±0.5° (IM2). The 3D magnitude of SE, preRE, postRE, IM1, and IM2 exceeded 2 mm in 18, 0, 3, 3, and 1 patients, respectively. No association were found between all positional uncertainties and body mass index, pain score, and treatment location (p > 0.05, Mann-Whitney test). There was a tendency of intrafraction motion to increase with overall treatment time; however, the correlation was not statistically significant (p > 0.05, Spearman rank correlation test). CONCLUSION: In spine SBRT using TM immobilization, CBCT and 4 DoF alignment correction, a minimum residual translational uncertainty was 2 mm. Shortening overall treatment time and 6 DoF positional correction may further reduce positional uncertainties.