Abstract P2-03-09: Tissue hemoglobin oxygen saturation measured by photoacoustic mammography correlates with microvasculature properties assessed by histological image analysis, a preliminary study
Authors: Fakhrejahani E , Torii M , Yamaga I , Asao Y , Kitai T , Kataoka M , Kanao S , Takada M , Shiina T , Toi M 
Published: August 2014

Abstract
Background: Photoacoustic mammography (PAM) is a new optical imaging technology with the potential of imaging tumor vasculature in breast cancer.The technique is applicable to the measurement of hemoglobin oxygen saturation (SO2).We have previously published an initial clinical result using a prototype machine (Canon Inc.,Tokyo, Japan) in breast cancer. We have also presented the morphological characteristics of tumor vessels analyzed by automated image processing during AACR annual meeting 2013. Here, we report the oxygenation data obtained by PAM in relation with histological assessment of tumor vasculature and hypoxia.
Methods: Forty-four breast lesions were evaluated by PAM in this IRB approved prospective study at Kyoto University Hospital, Japan. PAM evaluation was performed on both breasts whenever possible.SO2 was calculated in region of interest after the radiologist confirmed the signal was associated with the tumor location in MRI images. For the normal breast, signals obtained at the same depth of the tumor, were selected. Eighty-one histological sections from 20 available invasive carcinoma tissues at the time of this analysis were selected for immunohistochemical assessment of hypoxia by anti -carbonic anhydrase IX (CA IX) and tumor vasculature image analysis using anti-CD31. Five 0.5 mm2 area of each cancer and 3 area of normal mammary tissue associated with the same lesion were randomly selected from different sections. Total vascular area in each square was calculated by using Image Pro-Plus 7.0 software (Media Cybernetics, USA). Tumor-to-normal vascular area ratio (T/N VA) was calculated for each lesion as an index for tumor blood supply.
Results: Patients’ age ranged from 36 to 83 years old. Tumor associated signals were detected by PAM in 18 out of 20 lesions for which tissues were available for histological examination. SO2 in tumor was calculated 70.6% ±13.2 and 83.3% ±10.7 in the normal counterpart. While T/N VA ranged between 0.11 to 1.14, it was almost 3 times lower in lesions with CA IX positive cytoplasmic membrane staining (0.21 vs 0.7, p-value = 0.021 Mann-Whitney Test). Normalized tumor SO2 (tumor SO2/normal counterpart SO2) was significantly lower in the group with lower T/N VA (0.9 vs. 0.8,p-value = 0.045, Student T-test). To better evaluate the accuracy of PAM measurement in calculating SO2,3780 tumor-associated and 2835 normal microvessels were analyzed by image analysis software. Tumor associated vessels had significantly smaller area (p-value&lt;0.001) and vessels with irregular lumens were more frequent in tumor (76.5% vs 19.6% p-value &lt;0.001) compatible with lower SO2 in tumor areas.
Conclusion: Although the future result of our ongoing clinical studies of PAM measurement in breast cancer patients are more than necessary, the strong correlation between histological evaluation of hypoxia and angiogenesis with PAM measurement of oxygenation shows the promising prospective for clinical application of this new technology in breast cancer.
Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P2-03-09.