Use of 3D Spheroid Models for the Assessment of RT Response in Head and Neck Cancer.

Radiotherapy (RT) is a key player in the treatment of head and neck cancer (HNC). The RT response, however, is variable and influenced by multiple tumoral and tumor microenvironmental factors, such as human papillomavirus (HPV) infections and hypoxia. To investigate the biological mechanisms behind these variable responses, preclinical models are crucial. Up till now, 2D clonogenic and in vivo assays have remained the gold standard, although the popularity of 3D models is rising. In this study, we investigate the use of 3D spheroid models as a preclinical tool for radiobiological research by comparing the RT response of two HPV-positive and two HPV-negative HNC spheroid models to the RT response of their corresponding 2D and in vivo models. We demonstrate that HPV-positive spheroids keep their higher intrinsic radiosensitivity when compared to HPV-negative spheroids. A good correlation is found in the RT response between HPV-positive SCC154 and HPV-negative CAL27 spheroids and their respective xenografts. In addition, 3D spheroids are able to capture the heterogeneity of RT responses within HPV-positive and HPV-negative models. Moreover, we demonstrate the potential use of 3D spheroids in the study of the mechanisms underlying these RT responses in a spatial manner by whole-mount Ki-67 and pimonidazole staining. Overall, our results show that 3D spheroids are a promising model to assess the RT response in HNC.

Moderate dose escalation with volumetric modulated arc therapy improves outcome in rectal cancer.

BACKGROUND: Locally advanced rectal cancer is frequently treated with a long course of preoperative chemoradiotherapy. We investigated the effect of moderate dose escalation with volumetric modulated arc therapy (VMAT) up to 50 Gy in 25 fractions compared to 3D conformal radiotherapy (3D-CRT) to 45 Gy in 25 fractions in rectal cancer patients. Dose-volume parameters, acute toxicity, and complete response rates were compared. MATERIAL AND METHODS: For both groups, 65 patients were selected from our database through availability. Dose-volume parameters and acute toxicity data were compared using a Mann-Whitney U-test. Univariate and multivariate analyses correcting for tumor and nodal stage, distance to the mesorectal fascia and interval to surgery were used to compare complete response rates. RESULTS: Lower mean doses to the small and large bowel were observed in the VMAT group compared to the 3D-CRT group (21 Gy vs. 29 Gy [p < .001] and 26 Gy vs. 30 Gy [p = .002], respectively). Similar beneficial dose-volume parameters were observed for the bladder, sacrum and femoral heads. Furthermore, patients receiving VMAT experienced significantly less diarrhea, flatulence, non-infective cystitis, urinary frequency, dermatitis, and fatigue. In univariate analysis, a significant increase in complete response rate after moderate dose escalation with VMAT was observed (34% vs. 15%, p = .015). However, this did not remain significant when corrected for tumor and nodal stage, distance to the mesorectal fascia, and interval to surgery. CONCLUSIONS: Moderate dose escalation with VMAT resulted in superior dose-volume parameters compared to 3D-CRT, translating into lower acute toxicity. Additionally, improved tumor response after VMAT up to 50 Gy might contribute to a higher percentage of patients achieving a complete response.

Benefits of automated gross tumor volume segmentation in head and neck cancer using multi-modality information.

PURPOSE: Gross tumor volume (GTV) delineation for head and neck cancer (HNC) radiation therapy planning is time consuming and prone to interobserver variability (IOV). The aim of this study was (1) to develop an automated GTV delineation approach of primary tumor (GTVp) and pathologic lymph nodes (GTVn) based on a 3D convolutional neural network (CNN) exploiting multi-modality imaging input as required in clinical practice, and (2) to validate its accuracy, efficiency and IOV compared to manual delineation in a clinical setting. METHODS: Two datasets were retrospectively collected from 150 clinical cases. CNNs were trained for GTV delineation with consensus delineation as ground truth, with either single (CT) or co-registered multi-modal (CT + PET or CT + MRI) imaging data as input. For validation, GTVs were delineated on 20 new cases by two observers, once manually, once by correcting the delineations generated by the CNN. RESULTS: Both multi-modality CNNs performed better than the single-modality CNN and were selected for clinical validation. Mean Dice Similarity Coefficient (DSC) for (GTVp, GTVn) respectively between automated and manual delineations was (69%, 79%) for CT + PET and (59%,71%) for CT + MRI. Mean DSC between automated and corrected delineations was (81%,89%) for CT + PET and (69%,77%) for CT + MRI. Mean DSC between observers was (76%,86%) for manual delineations and (95%,96%) for corrected delineations, indicating a significant decrease in IOV (p < 10-5), while efficiency increased significantly (48%, p < 10-5). CONCLUSION: Multi-modality automated delineation of GTV of HNC was shown to be more efficient and consistent compared to manual delineation in a clinical setting and beneficial over a single-modality approach.

Hypoxia and Its Influence on Radiotherapy Response of HPV-Positive and HPV-Negative Head and Neck Cancer.

Head and neck squamous cancers are a heterogeneous group of cancers that arise from the upper aerodigestive tract. Etiologically, these tumors are linked to alcohol/tobacco abuse and infections with high-risk human papillomavirus (HPV). HPV-positive HNSCCs are characterized by a different biology and also demonstrate better therapy response and survival compared to alcohol/tobacco-related HNSCCs. Despite this advantageous therapy response and the clear biological differences, all locally advanced HNSCCs are treated with the same chemo-radiotherapy schedules. Although we have a better understanding of the biology of both groups of HNSCC, the biological factors associated with the increased radiotherapy response are still unclear. Hypoxia, i.e., low oxygen levels because of an imbalance between oxygen demand and supply, is an important biological factor associated with radiotherapy response and has been linked with HPV infections. In this review, we discuss the effects of hypoxia on radiotherapy response, on the tumor biology, and the tumor microenvironment of HPV-positive and HPV-negative HNSCCs by pointing out the differences between these two tumor types. In addition, we provide an overview of the current strategies to detect and target hypoxia.