Pre-treatment FMISO hypoxic volume is a significant prognostic factor for local control after irradiation of FaDu HNSCC xenografts

Objective: To investigate whether pre-treatment FMISO hypoxic tumour volume (HV) adds significant information about radiotherapy outcome in FaDu human squamous cell carcinoma (hSCC) in nude mice.
Materials and Methods: The hSCC cell line FaDu was transplanted subcutaneously into the hind leg of NMRI nude mice. Seventy animals entered the study at tumour volumes ranging from 165-343 mm³. [18F]fluoromisonidazole ([18F]FMISO)-PET scanning was performed under anesthesia (9% desflurane in 40% oxygen/air) on a dedicated animal PET scanner (MicroPET® P4, CTI Molecular Imaging Inc, measured attenuation correction, 11 MBq 18FMISO i.v., list mode acquisition for 30 min after 210 min p.i). The regions of interest (ROI) include the FMISO positive hypoxic volume, the mean, the maximum concentration (ROVER software, ABX GmbH, Radeberg, Germany). After an initial FMISO-PET (day 0) the tumours were stratified according to the median hypoxic volume (HV) for single dose irradiation with either 25 Gy (tumour control probability, TCP20) or 35 Gy (TCP80) under normal blood flow conditions using 200 kV X-rays (0.5 mm Cu, ~ 1.2 Gy min-1). The endpoint was time to local failure. Five animals are currently still in follow up.
Results: Tumour local control rate after irradiation with 25 Gy was lower than after irradiation with 35 Gy (22% vs. 69%, log rank p<0.0001). HV ranged from 38-353 mm³. Median HV was 112 mm³ (95%CI: 92; 128 mm³). In tumours with HV less than the median, local control was 33% after 25 Gy vs. 82% after 35 Gy (p=0.001) and in tumours with HV above the median 15% after 25 Gy vs. 53% after 35 Gy (p=0.0005). Multivariate Cox analysis revealed a significant effect of hypoxic volume treated either as a continuous (p=0.009) or a dichotomic variable (stratification by median HV) (p=0.039) when corrected for dose and tumour volume effects. Dose had a significant impact on hazard of recurrence (p<0.0005), whereas total tumour volume showed no effect (p=0.5).
Conclusions: Hypoxic volume is a significant predictor of tumour control after irradiation with high single doses in a single tumour line. This supports the hypothesis that pre-treatment FMISO-PET may provide useful information for heterogeneous radiation dose prescription in sub volumes of individual tumours. Confirmatory investigations using other tumour models and fractionated radiotherapy are warranted.
This work was performed within the 6th framework EU-project BioCare, proposal# 505785.