Elham Fakhrejahani

Antiangiogenesis therapy for breast cancer: an update and perspectives from clinical trials.

The development of new blood vessels is a crucial step in breast cancer growth, progression and dissemination, making it a promising therapeutic target. Breast cancer has a heterogeneous nature and the diversity of responsible angiogenic pathways between different tumors has been studied for many years. Inhibiting different targets in these pathways has been under investigation in preclinical and clinical studies for more than decades, among which antibody against vascular endothelial growth factor is the most studied. However, the clinical impact from antiangiogenic treatment alone or in combination with standard chemotherapeutic regimens has been relatively small till today. In this review, we summarize the most clinically relevant data from breast cancer treatment clinical trials and discuss safety and efficacy of common antiangiogenic therapies as well as biological predictive markers.

Tumor angiogenesis: Pericytes and maturation are not to be ignored

Angiogenesis, an essential component of tumor growth and survival, is regulated by complex interactions between several cell types and soluble mediators. Heterogeneous tumor vasculature originates from the collective effect of the nature of carcinoma and the complexity of the angiogenic network. Although the application of angiogenesis inhibitors in some types of cancers has shown clinical benefits, predictive markers to assess treatment effects have yet to be established. In this review, we focus on tumor vessel maturity as a potential marker for evaluating treatment response.

Visualization of tumor-related blood vessels in human breast by photoacoustic imaging system with a hemispherical detector array

Noninvasive measurement of the distribution and oxygenation state of hemoglobin (Hb) inside the tissue is strongly required to analyze the tumor-associated vasculatures. We developed a photoacoustic imaging (PAI) system with a hemispherical-shaped detector array (HDA). Here, we show that PAI system with HDA revealed finer vasculature, more detailed blood-vessel branching structures, and more detailed morphological vessel characteristics compared with MRI by the use of breast shape deformation of MRI to PAI and their fused image. Morphologically abnormal peritumoral blood vessel features, including centripetal photoacoustic signals and disruption or narrowing of vessel signals, were observed and intratumoral signals were detected by PAI in breast cancer tissues as a result of the clinical study of 22 malignant cases. Interestingly, it was also possible to analyze anticancer treatment-driven changes in vascular morphological features and function, such as improvement of intratumoral blood perfusion and relevant changes in intravascular hemoglobin saturation of oxygen. This clinical study indicated that PAI appears to be a promising tool for noninvasive analysis of human blood vessels and may contribute to improve cancer diagnosis.

Sinisan, a traditional Chinese medicine, attenuates experimental chronic pancreatitis induced by trinitrobenzene sulfonic acid in rats.

Background/purposeSinisan, a traditional Chinese medicine, is effective for the treatment of gastrointestinal disorders. In this study, we investigated the potential protective role of Sinisan against chronic pancreatitis (CP) in rats.MethodsCP was induced in rats by intrapancreatic injection of trinitrobenzene sulfonic acid (TNBS). Rats were randomly divided into a sham group, a TNBS-induced CP group and a Sinisan-treated group. Serum amylase and histological score were used to evaluate the severity of disease. The levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), cyclooxygenase-2 (COX-2), interleukin-10 (IL-10) and α-smooth muscle actin (α-SMA) were also measured in the three groups. Mechanical allodynia was measured with von Frey filaments. In addition, the protein levels of nerve growth factor (NGF) were measured in pancreatic tissues.ResultsAdministration of Sinisan significantly decreased the severity of CP. In the Sinisan-treated group, serum amylase, TNF-α, IL-1β, COX-2 and α-SMA levels were lower and the level of IL-10 was upregulated compared with the TNBS-induced CP group. Furthermore, treatment with Sinisan significantly, though not completely, attenuated the allodynia. Simultaneously NGF expression was also significantly downregulated in the Sinisan-treated group compared with the TNBS-induced CP group.ConclusionsSinisan could be an effective treatment modality for CP via its anti-inflammatory, anti-fibrotic and analgesic properties. It may be a promising drug candidate for the treatment of patients with CP.

Clinical Report on the First Prototype of a Photoacoustic Tomography System with Dual Illumination for Breast Cancer Imaging.

Photoacoustic tomography is a recently developed imaging modality that can provide high spatial-resolution images of hemoglobin distribution in tissues such as the breast. Because breast cancer is an angiogenesis-dependent type of malignancy, we evaluated the clinical acceptability of breast tissue images produced using our first prototype photoacoustic mammography (PAM) system in patients with known cancer. Post-excisionally, histological sections of the tumors were stained immunohistochemically (IHC) for CD31 (an endothelial marker) and carbonic anhydrase IX (CAIX) (a marker of hypoxia). Whole-slide scanning and image analyses were used to evaluate the tumor microvessel distribution pattern and to calculate the total vascular perimeter (TVP)/area for each lesion. In this clinical study, 42 lesions were primarily scanned using PAM preoperatively, three of which were reported to be benign and were excluded from statistical analysis. Images were produced for 29 out of 39 cancers (visibility rate = 74.4%) at the median depth of 26.5 (3.25–51.2) mm. Age, menopausal status, body mass index, history of neoadjuvant treatment, clinical stage and histological tumor angiogenesis markers did not seem to affect the visibility. The oxygen saturation level in all of the measured lesions was lower than in the subcutaneous counterpart vessels (Wilcoxon test, p value<0.001), as well as in the counterpart contralateral normal breast region of interest (ROI) (Wilcoxon test, p value = 0.001). Although the oxygen saturation level was not statistically significant between CAIX-positive vs. -negative cases, lesional TVP/area showed a positive correlation with the oxygen saturation level only in the group that had received therapy before PAM. In conclusion, the vascular and oxygenation data obtained by PAM have great potential for identifying functional features of breast tumors.

Photoacoustic mammography capable of simultaneously acquiring photoacoustic and ultrasound images

Abstract. We have constructed a prototype photoacoustic mammography system (PAM-02) capable of simultaneously acquiring photoacoustic (PA) and ultrasound (US) images. Each PA, US, and fused PA/US image can be acquired over a wide area of the breast using the scanning module of a US transducer, a PA detector, and optical prisms. The resolution of the PA images exhibits improvement from 2 to 1 mm compared to images acquired using our previous prototype. The maximum scan area of PAM-02 is 90 mm along the horizontal axis and 150 mm along the vertical axis. In a phantom experiment, the available depth was at least 45 mm. A representative example of the application of the PAM-02 prototype in clinical research at Kyoto University is presented and shows S-factor images, which are considered an approximation parameter related to hemoglobin saturation of tumor-related blood vessels. We confirmed the applicability of the system for anatomical and biological research.

Tumor microvasculature characteristics studied by image analysis: Histologically-driven angiogenic profile

Angiogenesis, a hallmark of cancer, has been studied to be a potential marker for diagnosis, prognosis and therapy in breast cancer. To evaluate tumor angiogenesis, histological assessment has been a common approach and counting tumor microvessels after visualizing them by immunohistochemistry has been in use for a long time. With recent advances in digital pathology and image analysis, other characteristics of tumor vasculature can also be evaluated. In this article we briefly review the potentials of image analysis in assessing tumor microvessel morphologically that might be helpful in defining a better angiogenesis marker than other common markers like vessel count.

Abstract P4-01-10: Development of photoacoustic vascular imaging system for breast cancer

Background: Tumor angiogenesis and hypoxia are associated with breast cancer growth and metastasis. Photoacoustic (PA) tomography is an optical imaging technology that visualizes distribution and oxygenation status of hemoglobin with high spatial resolution. Initially we developed a photoacoustic mammography (PAM) having a flat-shaped scanning detector that could detect breast tumors. Nevertheless, the flat-shaped detector array has the drawback of a limited view. Here we developed a novel PAM system with a hemispherical-shaped detector array (HDA), which enables us to identify microvasculatures non-invasively and allow the collection of nearly spatially isotropic three-dimensional reconstructed image of blood vessels. This non-invasive vascular imaging system may be able to characterize tumor angiogenesis and analyze the status of microcirculation. The aim of this study was to analyze the imaging findings of tumor-related vasculature in breast cancer patients. Patients and method: A PAM system with HDA has been generated in a cooperation project between Canon Inc., Japan, and Kyoto University. Twenty-two primary breast cancer patients, including 5 patients with non-invasive cancer and 17 patients with invasive cancer, diagnosed between December 2014 and December 2015 underwent the PAM imaging analysis. We also applied the breast deformation algorithm from the breast shape in a MRI image to that in a PA image in order to create a fusion image of the two modalities for the analysis. Features of peri- and intra-tumoral vasculature, and their oxygenation status were evaluated. The study protocol was approved by the institutional review board at Kyoto University Hospital (UMIN000012251). All patients provided informed consent to participate in this study. Results: The abnormal peri-tumoral vasculature was detected in 86% of all non-invasive and invasive disease cases. In invasive cancer cases, most tumor-related blood vessels were centripetally directed toward the tumor, and 93% of centripetal blood vessels appeared to be disrupted or rapidly narrowed at the tumor boundary. The centripetal blood vessel structure was frequently observed in invasive cancer compared with non-invasive cancer (61% vs 35%). PA images before and after preoperative chemotherapy were obtained in one case, where intra-tumoral blood vessels became finer after chemotherapy, reflecting normalization of intra-tumoral microcirculation induced by chemotherapy. Conclusions: A PAM system with HDA has provided a high-resolution vascular images of primary breast cancers. The morphological differences of peri-tumoral vasculature were observed between invasive disease and non-invasive disease. These results suggest the potential of PA imaging as a non-invasive tool to analyze tumor vasculature of human breast cancers and maybe be helpful for breast cancer diagnosis. (Acknowledgements) This work was partially supported by the Innovative Techno-Hub for Integrated Medical Bio-imaging Project of the Special Coordination Funds for Promoting Science and Technology from the Ministry of Education, Culture, Sports, Science, and Technology, Japan. Citation Format: Toi M, Asao Y, Takada M, Kataoka M, Endo T, Kawashima M, Yamaga I, Nakayama Y, Tokiwa M, Fakhrejahani E, Torii M, Kawaguchi-Sakita N, Kanao S, Matsumoto Y, Yagi T, Sakurai T, Togashi K, Shiina T. Development of photoacoustic vascular imaging system for breast cancer [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P4-01-10.

Abstract P4-03-03: Detection of the tumor vasculature and the hypoxic status of breast lesions using second-generation photoacoustic mammography: An exploratory study

Background: Tumor angiogenesis and hypoxia are associated with breast cancer growth and metastasis. Photoacoustic mammography (PAM) non-invasively visualizes hemoglobin distribution inside the breast by detecting thermoelastic waves from hemoglobin generated by the irradiation of a near-infrared laser pulse. Oxygen saturation (SO2) can be calculated using photoacoustic (PA) signals obtained by two laser pulses of different wavelengths. We further improved the spatial resolution of PAM by approximately 1 mm and enhanced detectability by using a high-sensitivity detector. This new PAM technique can obtain both PAM images and ultrasonography (US) images simultaneously. The aim of this study was to explore the clinical usefulness of this PAM technique. Patients and methods: Women who had breast lesions were eligible for this study. The participants9 lesions were measured using the new PAM technique before they began treatment. The PAM images were evaluated by 5 physicians. First, the lesions were identified using only the PAM images. Second, we used US or contrast-enhanced magnetic resonance images (CE-MRI) to identify the locations of the lesions. Next, we evaluated the photoacoustic (PA) signals based on their locations. Peri-tumoral PA signals were defined as linear signals that congregated in the peri-tumoral area, boundary PA signals were defined as peri-tumoral signals that were disrupted at the lesion9s boundaries, and intra-tumoral PA signals were defined as any significant PA signals inside the tumor. SO2 was illustrated using a color scale. The study protocol was approved by the institutional review board at Kyoto University Hospital, Japan (UMIN000007464). Results: PAM was performed on 48 breast lesions in 45 patients, including 36 invasive carcinoma lesions, 8 ductal carcinoma in situ (DCIS) lesions, and 4 benign lesions. Evaluations of PA signals according to the locations of the lesion, with confirmation from US or CE-MRI, were successfully performed for 38 lesions. Peri-tumoral PA signals were detected in 33 lesions (87%), disrupted boundary PA signals were detected in 30 lesions (79%), and intra-tumoral PA signals were detected in 25 lesions (66%). The detection rates for peri-tumoral, boundary and intra-tumoral PA signals were 94%, 87%, and 65% for invasive carcinoma, and 60%, 40%, and 80% for DCIS, respectively. Intra-tumoral PA signals tended to be weaker than peri-tumoral PA signals in invasive carcinoma lesions, and they often displayed a spotty rather than a linear shape. Intra-tumoral PA signals were observed to have lower SO2 levels than peri-tumoral PA signals in 95% of invasive carcinoma lesions and in 75% of DCIS lesions. Although peri-tumoral and boundary PA signals were also detected in a 38-mm fibroadenoma, the intra-tumoral PA signals displayed a diffuse pattern. Conclusions: We demonstrated that high spatial resolution and use in combination with US and CE-MRI facilitate the region-specific evaluation of PAM imaging. PAM could become a useful tool for the evaluation of the hypoxic status of tumors by enhancing its sensitivity. Citation Format: Takada M, Kawashima M, Kataoka M, Kanao S, Yamaga I, Torii M, Tokiwa M, Fakhrejahani E, Sakurai T, Asao Y, Haga H, Shiina T, Togashi K, Toi M. Detection of the tumor vasculature and the hypoxic status of breast lesions using second-generation photoacoustic mammography: An exploratory study. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P4-03-03.

Abstract P3-01-02: Clinical, anatomical and histological characteristics of breast lesions visualized by photoacoustic mammography; first clinical study in CK project

Background: Photoacoustic mammography (PAM) is an optical imaging technique potentially capable of imaging breast vasculature as well as measuring hemoglobin oxygen saturation(SO2) in focal breast lesions. We presented the initial observation on first 20 cases using the first generation PAM (PAM-01) prototype made by Canon Inc.(Tokyo, Japan) (spatial resolution:2mm) in SABCS 2013. Here we report the clinico-pathological characteristics of all cases recruited in the first clinical study in CK project (Kyoto University/Canon joint research project) between Aug 2010 and March 2012. Methods: 57 patients were recruited in IRB approved study at Kyoto University Hospital, Japan. Forty-two breast harboring lesions and when possible contralateral breasts were evaluated by PAM-01. Axial maximum intensity projection (MIP)s were obtained and signals from consecutive MIPs confirmed to be associated with the tumor location in MRI by an expert breast radiologist were considered to be the region of interest (ROI).The same depth was used for ROI in the normal breast as control. Histological sections from the widest area of the lesions were evaluated post-excisional by immunohistochemistry using anti-CD31 as endothelial marker and anti-carbonic anhydrase IX (CA IX) as hypoxia marker. Histological slides were scanned and divided into 1680 squares (0.84x0.84 mm2)(Hamamatsu Inc. Japan) for image analysis. Total vascular perimeter (TVP)-in mm- was calculated for all the histological section by using Image Pro-Plus 7.0 software (Media Cybernetics, USA). Tumor area was measured in mm2. TVP index was calculated as TVP/area. Results: Photoacoustic signal was detected in 30 lesions out of 42 at the depth of 26.8 ± 12.8 mm from which 80% were located superior to nipple. CA IX positive cases in comparison with CA IX negative cases significantly showed higher TVP index (p-value =0.028 Mann-Whitney Test ) suggesting more angiogenic profile of hypoxic tumors. However, lesions without any detectable signal were reported to have only a bigger mass size histologically (26.6 vs. 14.8 mm, p-value 0.28, Mann-Whitney Test) regardless of their TVP index or CA IX expression level. Moreover,SO2 was calculated 70.9% for signals located inside tumors and 85.5% for signals associated with subcutaneous vessels in the same breast (p-value Conclusion: This is the largest clinical study of PAM till today and the correlation between histological profile of hypoxia and tumor microvasculature with PAM signal visibility as well as tissue SO2 seems promising. However, the improvement of techniques and resolution is necessary to develop a more clinically applicable non-invasive functional breast imaging modality for analyzing breast tumor vasculature and hypoxia. Some of these improvements have taken place for the second generation PAM (PAM-02) which is now under a clinical evaluation study. Citation Format: Elham Fakhrejahani, Masae Torii, Yasufumi Asao, Iku Yamaga, Toshiyuki Kitai, Masako Kataoka, Shotaro Kanao, Masahiro Takada, Tsuyoshi Shiina, Masakazu Toi. Clinical, anatomical and histological characteristics of breast lesions visualized by photoacoustic mammography; first clinical study in CK project [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P3-01-02.