TY  - JOUR
AB  - Widespread metastasis is the major cause of death from melanoma and other types of cancer. At present, the dynamic aspects of the metastatic cascade remain enigmatic. The feasibility to track circulating melanoma cells deep within living intact organisms can greatly impact our knowledge on tumor metastasis, but existing imaging approaches lack the sensitivity, spatio-temporal resolution or penetration depth to capture flowing tumor cells over large fields of view within optically-opaque biological tissues. Vast progress with the development of optoacoustic tomography technologies has recently enabled two- and three-dimensional imaging at unprecedented frame rates in the order of hundreds of Hertz, effectively mapping up to a million image voxels within a single volumetric snapshot. Herein, we employ volumetric optoacoustic tomography for real-time visualization of passage and trapping of individual B16 melanoma cells in the whole mouse brain. Detection of individual circulating melanoma cells was facilitated by substituting blood with an artificial cerebrospinal fluid that removes the strong absorption background in the optoacoustic images. The approach can provide new opportunities for studying trafficking and accumulation of metastatic melanoma cells in different organs.
AU  - Dean-Ben, X.L.
AU  - Weidenfeld, I.
AU  - Degtyaruk, O.
AU  - Ntziachristos, V.
AU  - Stiel, A.-C.
AU  - Razansky, D.
C1  - 59682
C2  - 49011
CY  - Ste 800, 230 Park Ave, New York, Ny 10169 Usa
SP  - 441-446
TI  - Deep tissue volumetric optoacoustic tracking of individual circulating tumor cells in an in intracardially perfused mouse model.
JO  - Neoplasia
VL  - 22
IS  - 9
PB  - Elsevier Science Inc
PY  - 2020
SN  - 1522-8002
ER  - 

TY  - JOUR
AB  - Development of imaging methods capable of furnishing tumor-specific morphological, functional, and molecular information is paramount for early diagnosis, staging, and treatment of breast cancer. Ultrasound (US) and optoacoustic (OA) imaging methods exhibit excellent traits for tumor imaging in terms of fast imaging speed, ease of use, excellent contrast, and lack of ionizing radiation. Here, we demonstrate simultaneous tomographic whole body imaging of optical absorption, US reflectivity, and speed of sound (SoS) in living mice. In vivo studies of 4T1 breast cancer xenografts models revealed synergistic and complementary value of the hybrid imaging approach for characterizing mammary tumors. While neovasculature surrounding the tumor areas were observed based on the vascular anatomy contrast provided by the OA data, the tumor boundaries could be discerned by segmenting hypoechoic structures in pulse-echo US images. Tumor delineation was further facilitated by enhancing the contrast and spatial resolution of the SoS maps with a full-wave inversion method. The malignant lesions could thus be distinguished from other hypoechoic regions based on the average SoS values. The reported findings corroborate the strong potential of the hybrid imaging approach for advancing cancer research in small animal models and fostering development of new clinical diagnostic approaches.
AU  - Lafci, B.*
AU  - Mercep, E.
AU  - Herraiz, J.L.*
AU  - Deán-Ben, X.L.*
AU  - Razansky, D.
C1  - 60475
C2  - 49313
CY  - Ste 800, 230 Park Ave, New York, Ny 10169 Usa
SP  - 770-777
TI  - Noninvasive multiparametric charac-terization of mammary tumors with transmission-reflection optoacoustic ultrasound.
JO  - Neoplasia
VL  - 22
IS  - 12
PB  - Elsevier Science Inc
PY  - 2020
SN  - 1522-8002
ER  - 

TY  - JOUR
AB  - Integrins play an important role in tumor progression, invasion and metastasis. Therefore we aimed to evaluate a preclinical imaging approach applying ανβ3 integrin targeted hybrid Fluorescence Molecular Tomography/X-ray Computed Tomography (FMT-XCT) for monitoring tumor progression as well as early therapy response in a syngeneic murine Non-Small Cell Lung Cancer (NSCLC) model. Lewis Lung Carcinomas were grown orthotopically in C57BL/6 J mice and imaged in-vivo using a ανβ3 targeted near-infrared fluorescence (NIRF) probe. ανβ3-targeted FMT-XCT was able to track tumor progression. Cilengitide was able to substantially block the binding of the NIRF probe and suppress the imaging signal. Additionally mice were treated with an established chemotherapy regimen of Cisplatin and Bevacizumab or with a novel MEK inhibitor (Refametinib) for 2 weeks. While μCT revealed only a moderate slowdown of tumor growth, ανβ3 dependent signal decreased significantly compared to non-treated mice already at one week post treatment. ανβ3 targeted imaging might therefore become a promising tool for assessment of early therapy response in the future.
AU  - Ma, X.
AU  - van Phi, V.*
AU  - Kimm, M.A.*
AU  - Prakash, J.
AU  - Kessler, H.*
AU  - Kosanke, K.*
AU  - Feuchtinger, A.
AU  - Aichler, M.
AU  - Gupta, A.*
AU  - Rummeny, E.J.*
AU  - Eisenblätter, M.*
AU  - Siveke, J.*
AU  - Walch, A.K.
AU  - Braren, R.*
AU  - Ntziachristos, V.
AU  - Wildgruber, M.*
C1  - 50105
C2  - 42020
CY  - New York
SP  - 8-16
TI  - Integrin-targeted hybrid fluorescence molecular tomography/X-ray computed tomography for imaging tumor progression and early response in non-small cell lung cancer.
JO  - Neoplasia
VL  - 19
IS  - 1
PB  - Elsevier Science Inc
PY  - 2017
SN  - 1522-8002
ER  - 

TY  - JOUR
AB  - Diversity of the design and alignment of illumination and ultrasonic transducers empower the fine scalability and versatility of optoacoustic imaging. In this study, we implement an innovative high-resolution optoacoustic mesoscopy for imaging the vasculature and tissue oxygenation within subcutaneous and orthotopic cancerous implants of mice in vivo through acquisition of tomographic projections over 180° at a central frequency of 24 MHz. High-resolution volumetric imaging was combined with multispectral functional measurements to resolve the exquisite inner structure and vascularization of the entire tumor mass using endogenous and exogenous optoacoustic contrast. Evidence is presented for constitutive hypoxemia within the carcinogenic tissue through analysis of the hemoglobin absorption spectra and distribution. Morphometric readouts obtained with optoacoustic mesoscopy have been verified with high-resolution ultramicroscopic studies. The findings described herein greatly extend the applications of optoacoustic mesoscopy toward structural and multispectral functional measurements of the vascularization and hemodynamics within solid tumors in vivo and are of major relevance to basic and preclinical oncological studies in small animal models.
AU  - Chekkoury, A.
AU  - Nunes, A.
AU  - Gateau, J.
AU  - Symvoulidis, P.
AU  - Feuchtinger, A.
AU  - Bézière, N.
AU  - Ovsepian, S.V.
AU  - Walch, A.K.
AU  - Ntziachristos, V.
C1  - 49333
C2  - 41746
CY  - New York
SP  - 459-467
TI  - High-resolution multispectral optoacoustic tomography of the vascularization and constitutive hypoxemia of cancerous tumors.
JO  - Neoplasia
VL  - 18
IS  - 8
PB  - Elsevier Science Inc
PY  - 2016
SN  - 1522-8002
ER  - 

TY  - JOUR
AB  - The CCL2-CCR2 chemokine axis has an important role in cancer progression where it contributes to metastatic dissemination of several cancer types (e.g., colon, breast, prostate). Tumor cell-derived CCL2 was shown to promote the recruitment of CCR2(+)/Ly6C(hi) monocytes and to induce vascular permeability of CCR2(+) endothelial cells in the lungs. Here we describe a novel decoy protein consisting of a CCL2 mutant protein fused to human serum albumin (dnCCL2-HSA chimera) with enhanced binding affinity to glycosaminoglycans that was tested in vivo. The monocyte-mediated tumor cell transendothelial migration was strongly reduced upon unfused dnCCL2 mutant treatment in vitro. dnCCL2-HSA chimera had an extended serum half-life and thus a prolonged exposure in vivo compared with the dnCCL2 mutant. dnCCL2-HSA chimera bound to the lung vasculature but caused minimal alterations in the leukocyte recruitment to the lungs. However, dnCCL2-HSA chimera treatment strongly reduced both lung vascular permeability and tumor cell seeding. Metastasis of MC-38GFP, 3LL, and LLC1 cells was significantly attenuated upon dnCCL2-HSA chimera treatment. Tumor cell seeding to the lungs resulted in enhanced expression of a proteoglycan syndecan-4 by endothelial cells that correlated with accumulation of the dnCCL2-HSA chimera in the vicinity of tumor cells. These findings demonstrate that the CCL2-based decoy protein effectively binds to the activated endothelium in lungs and blocks tumor cell extravasation through inhibition of vascular permeability.
AU  - Roblek, M.*
AU  - Strutzmann, E.*
AU  - Zankl, C.*
AU  - Adage, T.*
AU  - Heikenwälder, M.
AU  - Atlic, A.*
AU  - Weis, R.*
AU  - Kungl, A.*
AU  - Borsig, L.*
C1  - 47759
C2  - 39467
CY  - New York
SP  - 49-59
TI  - Targeting of CCL2-CCR2-glycosaminoglycan axis using a CCL2 decoy protein attenuates metastasis through inhibition of tumor cell seeding.
JO  - Neoplasia
VL  - 18
IS  - 1
PB  - Elsevier Science Inc
PY  - 2016
SN  - 1522-8002
ER  - 

TY  - JOUR
AB  - Angiogenesis is a central cancer hallmark, necessary for supporting tumor growth and metastasis. In vivo imaging of angiogenesis is commonly applied, to understand dynamic processes in cancer development and treatment strategies. However, most radiological modalities today assess angiogenesis based on indirect mechanisms, such as the rate of contrast enhancement after contrast agent administration. We studied the performance of raster-scan optoacoustic mesoscopy (RSOM), to directly reveal the vascular network supporting melanoma growth in vivo, at 50 MHz and 100 MHz, through several millimeters of tumor depth. After comparing the performance at each frequency, we recorded, for the first time, high-resolution images of melanin tumor vasculature development in vivo, over a period of several days. Image validation was provided by means of cryo-slice sections of the same tumor after sacrificing the mice. We show how optoacoustic (photoacoustic) mesoscopy reveals a potentially powerful look into tumor angiogenesis, with properties and features that are markedly different than other radiological modalities. This will facilitate a better understanding of tumor's angiogenesis, and the evaluation of treatment strategies.
AU  - Omar, M.
AU  - Schwarz, M.
AU  - Soliman, D.
AU  - Symvoulidis, P.
AU  - Ntziachristos, V.
C1  - 43717
C2  - 36661
CY  - New York
SP  - 208-214
TI  - Pushing the optical Imaging limits of cancer with multi-frequency-band Raster-Scan Optoacoustic Mesoscopy (RSOM).
JO  - Neoplasia
VL  - 17
IS  - 2
PB  - Elsevier Science Inc
PY  - 2015
SN  - 1522-8002
ER  - 

TY  - JOUR
AB  - Classic histology still represents the gold standard in tumor tissue analytics. However, two-dimensional analysis of single tissue slides does not provide a representative overview of the inhomogeneous tumor physiology, and a detailed analysis of complex three-dimensional structures is not feasible with this technique. To overcome this problem, we applied multispectral fluorescence ultramicroscopy (UM) to the field of tumor analysis. Optical sectioning of cleared tumor specimen provides the possibility to three-dimensionally acquire relevant tumor parameters on a cellular resolution. To analyze the virtual UM tumor data sets, we created a novel set of algorithms enabling the fully automatic segmentation and quantification of multiple tumor parameters. This new postmortem imaging technique was applied to determine the therapeutic treatment effect of bevacizumab on the vessel architecture of orthotopic KPL-4 breast cancer xenografts at different time points. A significant reduction of the vessel volume, number of vessel segments, and branching points in the tumor periphery was already detectable 1 day after initiation of treatment. These parameters remained virtually unchanged in the center of the tumor. Furthermore, bevacizumab-induced vessel normalization and reduction in vascular permeability diminished the penetration behavior of trastuzumab-Alexa 750 into tumor tissue. Our results demonstrated that this new imaging method enables the three-dimensional visualization and fully automatic quantification of multiple tumor parameters and drug penetration on a cellular level. Therefore, UM is a valuable tool for cancer research and drug development. It bridges the gap between common macroscopic and microscopic imaging modalities and opens up new three-dimensional (3D) insights in tumor biology.
AU  - Dobosz, M.
AU  - Ntziachristos, V.
AU  - Scheuer, W.*
AU  - Strobel, S.*
C1  - 31383
C2  - 34450
CY  - New York
SP  - 1-13
TI  - Multispectral fluorescence ultramicroscopy: Three-dimensional visualization and automatic quantification of tumor morphology, drug penetration, and antiangiogenic treatment response.
JO  - Neoplasia
VL  - 16
IS  - 1
PB  - Elsevier Science Inc
PY  - 2014
SN  - 1522-8002
ER  - 

TY  - JOUR
AB  - Searching for new strategies to bypass apoptosis resistance, we investigated the potential of the Smac mimetic BV6 in Jurkat leukemia cells deficient in key molecules of the death receptor pathway. Here, we demonstrate for the first time that Smac mimetic primes apoptosis-resistant, FADD- or caspase-8-deficient leukemia cells for TNFα-induced necroptosis in a synergistic manner. In contrast to TNFα, Smac mimetic significantly enhances CD95-induced apoptosis in wild-type but not in FADD-deficient cells. Interestingly, Smac mimetic- and TNFα-mediated cell death occurs without characteristic features of apoptosis (i.e., caspase activation, DNA fragmentation) in FADD-deficient cells. By comparison, Smac mimetic and TNFα trigger activation of caspase-8, -9, and -3 and DNA fragmentation in wild-type cells. Consistently, the caspase inhibitor zVAD.fmk fails to block Smac mimetic- and TNFα-triggered cell death in FADD- or caspase-8-deficient cells, while it confers protection in wild-type cells. By comparison, necrostatin-1, an RIP1 kinase inhibitor, abolishes Smac mimetic- and TNFα-induced cell death in FADD- or caspase-8-deficient. Thus, Smac mimetic enhances TNFα-induced cell death in leukemia cells via two distinct pathways in a context-dependent manner: it primes apoptosis-resistant cells lacking FADD or caspase-8 to TNFα-induced, RIP1-dependent and caspase-independent necroptosis, whereas it sensitizes apoptosis-proficient cells to TNFα-mediated, caspase-dependent apoptosis. These findings have important implications for the therapeutic exploitation of necroptosis as an alternative cell death program to overcome apoptosis resistance.
AU  - Laukens, B.*
AU  - Jennewein, C.*
AU  - Schenk, B.*
AU  - Vanlangenakker, N.*
AU  - Schier, A.*
AU  - Cristofanon, S.*
AU  - Zobel, K.*
AU  - Deshayes, K.*
AU  - Vucic, D.*
AU  - Jeremias, I.
AU  - Bertrand, M.J.*
AU  - Vandenabeele, P.*
AU  - Fulda, S.*
C1  - 6031
C2  - 29219
SP  - 971-979
TI  - Smac mimetic bypasses apoptosis resistance in FADD- or caspase-8-deficient cells by priming for tumor necrosis factor α-induced necroptosis.
JO  - Neoplasia
VL  - 13
IS  - 10
PB  - Neoplasia Press
PY  - 2011
SN  - 1522-8002
ER  - 

TY  - JOUR
AB  - The selenoenzyme glutathione peroxidase 4 (GPx4) has been described to control specific cyclooxygenases (COXs) and lipoxygenases (LOXs) that exert substantiated functions in tumor growth and angiogenesis. Therefore, we hypothesized a putative regulatory role of GPx4 during tumor progression and created transformed murine embryonic fibroblasts with inducible disruption of GPx4. GPx4 inactivation caused rapid cell death in vitro, which could be prevented either by lipophilic antioxidants or by 12/15-LOX-specific inhibitors, but not by inhibitors targeting other LOX isoforms or COX. Surprisingly, transformed GPx4(+/-) cells did not die when grown in Matrigel but gave rise to tumor spheroids. Subcutaneous implantation of tumor cells into mice resulted in knockout tumors that were indistinguishable in volume and mass in comparison to wild-type tumors. However, further analysis revealed a strong vascular phenotype. We observed an increase in microvessel density as well as a reduction in the number of large diameter vessels covered by smooth muscle cells. This phenotype could be linked to increased 12/15-LOX activity that was accompanied by an up-regulation of basic fibroblast growth factor and down-regulation of vascular endothelial growth factor A protein expression. Indeed, pharmacological inhibition of 12/15-LOX successfully reversed the tumor phenotype and led to "normalized" vessel morphology. Thus, we conclude that GPx4, through controlling 12/15-LOX activity, is an important regulator of tumor angiogenesis as well as vessel maturation.
AU  - Schneider, M.
AU  - Wortmann, M.*
AU  - Mandal, P.K.
AU  - Jannasch, K.*
AU  - Alves, F.*
AU  - Strieth, S.*
AU  - Conrad, M.
AU  - Beck, H.*
C1  - 5985
C2  - 27980
SP  - 254-263
TI  - Absence of glutathione peroxidase 4 affects tumor angiogenesis through increased 12/15-lipoxygenase activity.
JO  - Neoplasia
VL  - 12
IS  - 3
PB  - Neoplasia Press, Inc.
PY  - 2010
SN  - 1522-8002
ER  - 

TY  - JOUR
AB  - Optical imaging of breast cancer has been considered for detecting functional and molecular characteristics of diseases in clinical and preclinical settings. Applied to laboratory research, photonic investigations offer a highly versatile tool for preclinical imaging and drug discovery. A particular advantage of the optical method is the availability of multiple spectral bands for performing imaging. Herein, we capitalize on this feature to demonstrate how it is possible to use different wavelengths to offer internal controls and significantly improve the observation accuracy in molecular imaging applications. In particular, we show the independent in vivo detection of cysteine proteases along with tumor permeability and interstitial volume measurements using a dual-wavelength approach. To generate results with a view toward clinically geared studies, a transgenic Her2/neu mouse model that spontaneously developed mammary tumors was used. In vivo findings were validated against conventional ex vivo tests such as histology and Western blot analyses. By correcting for biodistribution parameters, the dual-wavelength method increases the accuracy of molecular observations by separating true molecular target from probe biodistribution. As such, the method is highly appropriate for molecular imaging studies where often probe delivery and target presence are not independently assessed. On the basis of these findings, we propose the dual-wavelength/normalization approach as an essential method for drug discovery and preclinical imaging studies.
AU  - Baeten, J.
AU  - Haller, J.*
AU  - Shih, H.*
AU  - Ntziachristos, V.
C1  - 733
C2  - 26077
SP  - 220-227
TI  - In vivo investigation of breast cancer progression by use of an internal control.
JO  - Neoplasia
VL  - 11
IS  - 3
PB  - Neoplasia Press
PY  - 2009
SN  - 1522-8002
ER  - 

TY  - JOUR
AB  - BACKGROUND AND AIMS: Although both cancer and stellate cells (PSCs) secrete proangiogenic factors, pancreatic cancer is a scirrhous and hypoxic tumor. The impact of cancer-PSCs interactions on angiogenesis was analyzed. METHODS: Expression of periostin, CD31, and alpha-smooth muscle actin was assessed by immunohistochemistry. Human PSCs and cancer cells were cultivated under normoxia and hypoxia alone, or in coculture, to analyze the changes in their angiogenic and fibrogenic attributes, using enzyme-linked immunosorbent assay, immunoblot, and quantitative polymerase chain reaction analyses and growth of cultured endothelial cells in vitro. RESULTS: On the invasive front of the activated stroma, PSCs deposited a periostin-rich matrix around the capillaries in the periacinar spaces. Compared with the normal pancreas, there was a significant reduction in the microvessel density in chronic pancreatitis (five-fold, P < .001) and pancreatic cancer (four-fold, P < .01) tissues. In vitro, hypoxia increased PSCs' activity and doubled the secretion of periostin, type I collagen, fibronectin, and vascular endothelial growth factor (VEGF). Cancer cells induced VEGF secretion of PSCs (390 +/- 60%, P < .001), whereas PSCs increased the endostatin production of cancer cells (210 +/- 14%, P < .001) by matrix metalloproteinase-dependent cleavage. In vitro, PSCs increased the endothelial cell growth, whereas cancer cells alone, or their coculture with PSCs, suppressed it. CONCLUSIONS: Although PSCs are the dominant producers of VEGF and increase endothelial cell growth in vitro, in the peritumoral stroma, they contribute to the fibrotic/hypoxic milieu through abnormal extracellular matrix deposition and by amplifying endostatin production of cancer cells.
AU  - Erkan, M.*
AU  - Reiser-Erkan, C.*
AU  - Michalski, C.W.*
AU  - Deucker, S.*
AU  - Sauliunaite, D.*
AU  - Streit, S.*
AU  - Esposito, I.
AU  - Friess, H.*
AU  - Kleeff, J.*
C1  - 1021
C2  - 26904
SP  - 497-508
TI  - Cancer-stellate cell interactions perpetuate the hypoxia-fibrosis cycle in pancreatic ductal adenocarcinoma.
JO  - Neoplasia
VL  - 11
IS  - 5
PB  - Neoplasia Press
PY  - 2009
SN  - 1522-8002
ER  -