Intratumoral Vessels Research Articles

Abstract PR04: Intravital imaging of cytolytic activity reveals how poor tumor vascularization and hypoxia limit tumor rejection

Abstract Background: Immunotherapies that harness cytotoxic T lymphocytes (CTLs) to eradicate solid tumors, melanoma in particular, have re-emerged as potent clinical options in recent years. Improving the dispersion and cytotoxicity of intratumoral CTLs may further benefit patients. One of the obstacles to successful treatment is thought to be poor vascularization of tumors, which leads to sub-optimal tumor infiltration. In contrast, the effects of disrupted vascularization and the hypoxia it introduces on CTL migration and function within tumors have not been investigated. Methods: To study whether tumor vascularity can shape anti-tumor CTL responses, we combined immunohistochemistry of human melanoma samples with dynamic two-photon imaging within B16 melanoma tumors expressing ovalbumin (OVA) in live mice. Results: In patients, CD8 T cells were twofold denser around peripheral vessels than around deep vessels, and fourfold denser than in avascular areas. This pattern was clearest in poorly-infiltrated tumors. Correspondingly, in mice, adoptively transferred OT-I CTLs, specific for OVA, preferentially infiltrated B16-OVA tumors through peripheral ICAM1+ vessels. CTLs remained initially at their entry areas, crawling vigorously only within 50µm of flowing blood vessels. These dense CTLs swarms efficiently cleared perivascular areas of tumor cells but left avascular areas intact. In contrast, Foxp3+ regulatory T cells could venture farther away from blood vessels. Target killing was antigen-specific, contact-dependent and perforin-mediated, and required multiple CTLs to engage one target cell to be effective. Several findings converge to suggest that perivascular CTL confinement is oxygen-dependent: occlusion of intratumoral blood vessels resulted in instantaneous and reversible arrest of CTLs. Immunohistology indicated that CTLs avoided hypoxic tumor areas. Hypoxia and mitochondrial uncoupling interfered with CTL migration in vitro and, finally, highly vascular tumors grown in a matrix containing the pro-angiogenic factor bFGF were more easily rejected after CTL injection, displayed delayed growth in untreated mice but were not affected in immunodeficient mice. Although such normalized tumors attracted normal numbers of CTLs, these cells migrated more freely within the tumors. Conclusions: We propose that CTLs depend on oxygen for intratumoral locomotion and cytolysis. Incomplete vascularization promotes patchy infiltration and tumor escape; tumor immunotherapy may thus benefit from an extensive and functional vasculature. Citation Format: Guy Shakhar, Yoav Manaster, Tali Feferman. Intravital imaging of cytolytic activity reveals how poor tumor vascularization and hypoxia limit tumor rejection. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Angiogenesis and Vascular Normalization: Bench to Bedside to Biomarkers; Mar 5-8, 2015; Orlando, FL. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl):Abstract nr PR04.

Blood vessels and cancer much more than just angiogenesis.

The availability of oxygen and nutrients supplied by the vasculature is crucial for both normal and neoplastic cells, however, the nature of the intratumour vessels and their relationship to the neoplastic cells has been debated for centuries.

Transdermal drug targeting and functional imaging of tumor blood vessels in the mouse auricle

Subcutaneously growing tumors are widely utilized to study tumor angiogenesis and the efficacy of antiangiogenic therapies in mice. To additionally assess functional and morphologic alterations of the vasculature in the periphery of a growing tumor, we exploited the easily accessible and hierarchically organized vasculature of the mouse auricle. By site-specific subcutaneous implantation of a defined preformed mouse B16/F0 melanoma aggregate, a solid tumor nodule developed within 14 d. Growth of the tumor nodule was accompanied by a 4-fold increase in its perfusion as well as a 2- to 4-fold elevated diameter and perfusion of peripheral blood vessels that had connected to the tumor capillary microvasculature. By transdermal application of the anticancer drug bortezomib, tumor growth was significantly diminished by about 50% without provoking side effects. Moreover, perfusion and tumor microvessel diameter as well as growth and perfusion of arterial or venous blood vessels supplying or draining the tumor microvasculature were decreased under these conditions by up to 80%. Collectively, we observed that the progressive tumor growth is accompanied by the enlargement of supplying and draining extratumoral blood vessels. This process was effectively suppressed by bortezomib, thereby restricting the perfusion capacity of both extra and intratumoral blood vessels.

Solitary fibrous tumor of the pleura: Can computed tomography features help predict malignancy? A series of 56 patients with histopathological correlates

To identify computed tomography (CT) predictors of malignancy, from a retrospective study of preoperative CT scans of patients with solitary fibrous tumors (SFT) of the pleura. The CT scans of 56patients with histopathologically confirmed SFT (33women and 23men; mean age, 60years) who underwent surgery between December2004 and November2012 were retrospectively analyzed by three radiologists working in consensus, blinded to the final histological diagnosis. SFT was asymptomatic and incidentally discovered in 22patients (45.8%). Resection specimen analysis (R0 resection in all cases) revealed that 23tumors (41%) were malignant. The CT features, which significantly differed between malignant and benign SFTs were tumor size (P=0.002) with a discriminative threshold value of 10cm, tumor heterogeneity before (P=0.02) and after (P=0.03) intravenous administration of iodinated contrast material, presence of intratumoral hydric attenuation areas (P=0.01), pleural effusion (P=0.01), measurable intratumoral vessels (P=0.02), hypervascularization with visible intratumoral vessels and/or marked enhancement (P=0.001). Presence of intratumoral calcifications (P=0.2) and maximum post-contrast enhancement value (P=0.6) were not significantly different between the two groups. A size greater than or equal to 10cm, hypervascularization, attenuation heterogeneity and association with pleural effusion are individual variables that suggest malignant SFT on CT.

A87: Proliferative activity, lymphatic and blood vessel density in different clinical stages of melanoma

Cutaneous melanoma is one of the most aggressive human neoplasms that can quickly metastasize to regional lymph nodes. Currently, prognosis is determined by measuring tumor thickness but more reliable markers for metastatic spread are urgently needed. It is well known that tumors require a microvasculature development in order to grow and metastasize. Angiogenesis and lymphangiogenesis play an important role not only in the tumor growth, but also in the tumor metastasis. As malignancy is a disorder of cellular growth control, the assessment of cell proliferation rates in tumors has intuitive appeal as a prognostic marker. One such biomarker is Ki-67, a cell cycle dependent protein. Recent reports related to its role as a prognostic factor have been contradictory. The purpose of this study was to investigate lymphangiogenesis, angiogenesis and tumor tissue proliferative activity depending on the clinical stages of melanoma. The objects of the study were the samples of tumor tissue obtained during surgical treatment of cutaneous malignant melanoma. To analyze tumor angiogenesis, lymphangiogenesis and proliferation, we performed immunostains of the primary melanomas for the vascular marker CD34, for the lymphatic-specific markers LYVE-1, D2 -40 (Podoplanin) and marker of proliferation – Ki-67. Samples of tumor tissue from 40 patients with IA, IB, IIA, IIB, IIIC melanoma stages were fixed in 10% neutral formalin, processed by standard histological techniques and embedded in paraffin. All steps of the immunohistochemical reaction were performed by using BENCHMARK/XT slide stainer (Ventana). Determination of blood vessel volume density revealed its growth an average of 2 times in peritumoral areas. Similar data were obtained about the location lymphatic vessels. Greater content peritumoral blood and lymphatic vessels, then intratumoral could be detected in all stages of melanoma. In addition more significantly greater volume density both intratumoral and peritumoral blood vessels, than lymphatic vessels was found. A greater degree expression of endothelium lymphatic vessels markers Podoplanin, compared with the LYVE-1 was shown. It was noted an increasing volume density of the peritumoral blood and lymphatic vessels in primary tumors with expansion clinical stage melanoma. Three levels of proliferative activity of tumor tissue were determined (low, an average degree and high Ki-67 expression). Results of the analysis have shown, that the high proliferative activity corresponded to significant content of blood and lymphatic vessels. High level of these markers was observed in patients with regional lymph nodes metastases. In recent years there have been appeared publications suggesting that lymphatic vessel density (particularly in a peritumoral location) and lymphatic vessel invasion are predictors of sentinel node metastasis and poorer survival. It was noted, that “larger, carefully conducted and well-reported studies that confirm these preliminary findings are required before it would be appropriate to recommend the routine application of costly and time-consuming immunohistochemistry for lymphatic markers in the routine clinical assessment of primary cutaneous melanomas”. In our opinion, one must consider not only the lymphatic vessels density, but also blood vessels, and the tumor tissue proliferative activity. Conclusion This study has shown that the complex markers of lymphangiogenesis (Podoplanin), angiogenesis (CD34) and proliferation (Ki-67) may be predictors of high risk early melanoma metastasis.

T19: The immune and vascular components of microenvironment in prognosis of the endometrioid endometrial adenocarcinoma

Background Endometrioid endometrial adenocarcimoma is the most frequent gynecologic malignancy in the western world. It is well known that inflammation plays a key role in the initiation and development of cancer as well as promotes both specific and innate immune response. There is no information on the status of the local anti-tumor cellular immunity in endometrioid adenocarcinoma of the corpus uteri microenvironment in the Russian and English language medical scientific literature. Various studies have shown that high intratumor microvessel density in endometrial cancer is associated with advanced stage, increased risk of recurrent disease and with poor prognosis. Vascular endothelial growth factor (VEGF) is a glycoprotein with potent mitogenic activity. It has been demonstrated to be a key component in the promotion of angiogenesis. Materials and methods There were singlet 24 cases of endometrioid adenocarcinoma with retrospectively known good (survival) and poor (death) outcomes. All cases were immunohistochemically stained for CD3, CD20, CD57, CD68, S100, CD34 and VEGF, expression of was evaluated by IHC-profiler. The number and the area of vessels was count in “hot spot” zones and calculated per 1 mm. For compare of the characteristics of signs was used comparison of two independent samples – U-Mann–Whitney test. The level of statistical significance was accepted by p Results T-lymphocytes cells were situated predominantly in desmoplastic stroma of the tumor. CD3 expression was significant higher in the group of good outcome (p = 0.00042), B-lymphocytes cells were predominantly in epithelial component of tumor. CD20 was expressed in endometriod adenocarcinoma with good and poor outcome without statistically significant difference (p = 0.976). NK-lymphocytes were predominantly in peritumoral stroma. CD57 expression was statistically significant higher in the group of good outcome (p = 0.000307). Tumor infiltrating macrophages can were frequent in tumor stroma than in tumor islets. CD68 expression cells number was higher in the group with good outcome (p = 0.000219). Dendric cells were predominantly in epithelial component of tumor. S100+ cells number did not have any difference (p = 0.105). Inside the tumor stroma, among pseudo-glands or solid beaches there was a positive reaction for anti-CD34 of numerous microvessels together with groups of endothelial cells (“hot spot”). The number of intratumoral vessels in patients with poor outcome was 141.9 ± 24.7 vessels/mm2, in good outcome patients – 93.8 ± 28.7 vessels/mm2. The number of the intratumoral vessels in this groups was with significant statistical different (p = 0.0014). The area of intratumoral vessels in good outcome cases was 2799.7 ± 535.7 μm2, poor outcome cases – 5455,2 ± 448,8 μm2. It was significant statistical different (p = 0.000014). VEGF was expressed in the cytoplasm of normal endometrial cells, neoplastic ones, endothelial cells and stroma. VEGF represents a mitogen for endothelial cells that induces the formation of lumen’s vessels and also increases vascular permeability and protein extravasation. Expression of VEGF by cancer cells had significant statistical difference in this groups of patients (p = 0.0411). Conclusion There was a significant reduction in T, NK-cells and macrophages from patients with poor outcome indicates the development of local immune deficiency that could certainly plays an important role in tumor progression endometrioid carcinoma. Our study demonstrates that the number and the area of vessels, as part of angiogenesis, represent an important prognostic factor in outcome of endometrioid endometrial adenocarcinoma. VEGF expression also is an angiogenic factor which plays a great role in the prognosis of this tumor.

Abstract B84: Live imaging reveals hypoxia to be a limiting factor in CTL-mediated tumor rejection

Abstract Background: One of the obstacles to successful immunotherapy by CTLs is thought to be poor vascularization of tumors that leads to sub-optimal tumor infiltration. In contrast, the effects of disrupted vascularization and the hypoxia it introduces on CTL migration and function within tumors have not been investigated. Methods: To study whether tumor vascularity can shape anti-tumor CTL responses, we combined immunohistochemistry of human melanoma samples with dynamic two-photon imaging within B16-OVA tumors in live mice. Results: In patients, CD8 T cells were twofold denser around peripheral vessels than around deep vessels, and fourfold denser than in avascular areas. This pattern was clearest in poorly-infiltrated tumors. Correspondingly, in mice, OT-I CTLs preferentially infiltrated B16-OVA tumors through peripheral ICAM1+ vessels. CTLs remained initially at their entry areas, crawling vigorously only within 50 µm of flowing blood vessels. Gradually, CTLs formed dense perivascular swarms, producing locally-high CTL-to-target ratios at their advancing front. These swarms advanced through the tumors, leading to patchy elimination of tumor cells. Target killing was antigen-specific, contact-dependent and perforin-mediated, and required several CTLs to engage one target cells to be effective. Several findings suggest that CTL confinement to perivascular areas is oxygen-dependent: occlusion of intratumoral blood vessels resulted in immediate arrest of CTLs. Immunohistology indicated that CTLs avoided hypoxic tumor areas. Hypoxia and mitochondrial uncoupling interfered with CTL migration in vitro and, finally, highly vascular tumors grown in a matrix containing pro-angiogenic factors were more easily rejected after CTL injection, displayed delayed growth in untreated mice but were not affected in immunodeficient mice. Conclusions: We suggest that CTLs depend on oxygen for intratumoral locomotion and cytolysis. Incomplete vascularization promotes patchy infiltration and tumor escape; tumor immunotherapy may thus benefit from normal and extensive tumor angiogenesis Citation Format: Guy Shakhar, Tali Feferman, Yoav Manaster, Anat Hutzler. Live imaging reveals hypoxia to be a limiting factor in CTL-mediated tumor rejection. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B84.

Abstract B06: Imaging regulatory mechanisms that limit intratumoral CTL function in a mouse melanoma model

Abstract Introduction: Immunotherapies that harness CTLs to eradicate solid tumors, melanoma in particular, have re-emerged as potent clinical options in recent years. Improving the dispersion and cytotoxicity of intratumoral CTLs may further benefit patients. To gain insight into the regulatory processes that limit CTL activity in melanoma we investigated the influence of PD-1 signaling and Tregs on CTL behavior. Methodology: To mimic adoptive cell therapy we transferred mice bearing B16.OVA melanoma tumors with OT-I CTLs. To study Treg-mediated regulation, we used FOXP3-GFP-LuciDTR-4 host mice, in which 95% of the cells expressing Foxp3 could be depleted using diphtheria toxin. To study PD-1 mediated regulation we injected anti PD-1 to WT host mice. Tumor rejection was monitored for 2-5 weeks. Two-photon imaging of live mice was used to follow the behavior of CTLs and Tregs within tumors. Time-lapse confocal microscopy was used to follow the influence of PD-1 signaling on the behavior of CTLs and their interaction with B16-Ova cells. Results: Tumor rejection was closely regulated by Tregs as tumors were densely infiltrated by these cells and their depletion dramatically accelerated tumor rejection. Interactions between CTLs and Tregs seemed indirect, as they did not form conjugates. Following their transfer, antigen-specific OT-I CTLs, as well as polyclonal CTLs, entered murine tumors through selected ICAM-decorated intratumoral vessels. As CTLs accumulated, Treg numbers in the tumors dropped and CTLs could be observed conjugating tumor cells and killing them. Treatment with anti PD-1 increased CTL velocities and accelerated killing of B16.OVA cells in vitro, and was mirrored in vivo by enhanced tumor rejection. Whereas CTL migration and cytolysis was limited to oxygenated tumor areas within 50 µm of flowing blood vessels, Tregs exhibited higher velocities, were not affected by the proximity of blood vessels, and penetrated deeper into avascular areas. Conclusions: Our findings suggest that regulatory mechanisms influence CTL efficiency partly by affecting their migration. CTLs, more than Tregs, depend on oxygen for intratumoral locomotion and cytolysis. This disparity allows tumor cells to escape CTL lysis in deep hypoxic tumor niches where high Treg to CTL ratios favor their survival. Thus, differential motility and dependence on oxygen among CTLs and Tregs can dictate the efficiency of CTL-based therapies of melanoma. Citation Format: Tali Feferman, Natalia G. Zabolinsky, Yoav Manaster, Anat Hutzler, Guy Shakhar. Imaging regulatory mechanisms that limit intratumoral CTL function in a mouse melanoma model. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B06.

Anatomic features of choroidal naevi: Swept‐source optical coherence tomography vs Enhanced depth imaging tomography. Preliminary results in 31 patients

PurposeTo assess the anatomic retinal and choroidal features of choroidal naevi using swept‐source optical coherence tomography (SS‐OCT) and enhanced‐depth optical coherence tomography (EDI‐OCT).DESIGNObservational case series.MethodsPatients with choroidal lesions underwent clinical examination, B‐scan ultrasound and imaging with SS‐OCT and EDI‐OCT. Location, dimensions, clinical and OCT retinal and choroidal features were recorded. Descriptive statistics were used.ResultsCase series included 31 patients. 27/31 naevi imaged were melanotic and 4/31 amelanotic with a an overall median thickness of 0.7 mm. Naevus configuration was plateau in 17/31 cases, dome in 10/31 cases and mixed in 4/31 cases. RPE and photoreceptor layer disruption were noted in 14/31 cases and 13/31 had no retinal changes. Subretinal fluid was noted in 6/31 cases. Bruch`s membrane was found intact in 26/31 cases on both modalities. Intrinsic hyperreflectivity was noted in 29/31 cases on EDI‐OCT and 30/31 cases on SS‐OCT with less optical shadowing. The posterior margin of the naevus was visualized in 11/31 cases with SS‐OCT and in 6/31 cases with EDI‐OCT. Intratumour vessels were visualized in 28/31 cases with SS‐OCT and 23/31 cases with EDI‐OCT. For both modalities choriocapillaris appeared compressed and abnormal in 20/31 cases.ConclusionsThese preliminary results indicate that imaging of choroidal naevi with SS‐OCT enables better visualization of intratumour vessels and the posterior margin.

A Novel Model of Metastatic Conjunctival Melanoma in Immune-Competent Mice.

Conjunctival melanoma (CM) is an ocular surface tumor that can lead to fatal metastases. Patients developing, tumor-associated lymphangiogenesis have a significantly increased risk of metastatic disease, because tumor spread primarily occurs via lymphatic vessels to the draining lymph node. Here, we describe a novel immune-competent mouse model of CM that displays tumor-associated lymphangiogenesis with development of metastatic tumors. C57BL/6N mice received C57BL/6N-derived dermal melanoma cells (hepatocyte growth factor [HGF] cyclin dependent kinase-4 [Cdk4]+) or B16F10 via subconjunctival injection. A clinical score quantified primary tumor growth and metastases were identified by macroscopic examination of the draining lymph nodes, lung, and spleen. Confirmation of tumors and metastases was achieved by immunohistochemical staining for markers of pigmented cells (tyrosinase related protein-2 [TRP2]) and S-100, and of cell proliferation (Ki67). The intra- and peritumoral CD31+ blood and lymphatic vessel endothelium hyaluronan receptor-1 (LYVE-1)+ lymphatic vessels were quantified immunohistochemically. All mice rapidly developed aggressive TRP2+, S100+, and Ki67+ CM. Metastatic tumors were found in the lymph node (9%) and lung (6%) of HGF-Cdk4(R24C)-treated mice and in the spleen (8%) and lung (17%) of B16F10-treated mice. The amount of peri- and intratumoral blood vessels was significantly increased compared with lymphatic vessels. This CM model in immune-competent animals offers new possibilities to study the pathobiology of tumor growth, invasion, and mechanisms of metastatic tumor spread, and provides a robust model to explore new immune-based and antilymphangiogenic treatment modalities of this malignancy.

Multidetector Computed Tomography Features in Differentiating Exophytic Renal Angiomyolipoma from Retroperitoneal Liposarcoma: A Strobe-Compliant Observational Study.

This study aims to evaluate the multidetector computed tomography (CT) imaging features in differentiating exophytic renal angiomyolipoma (AML) from retroperitoneal liposarcoma.We retrospectively enrolled 42 patients with confirmed exophytic renal AML (31 patients) or retroperitoneal liposarcoma (11 patients) during 8 years period to assess: renal parenchymal defect at site of tumor contact, supply from branches of renal artery, tumoral vessel extending through the renal parenchyma, dilated intratumoral vessels, hemorrhage, non–fat-containing intratumoral nodules with postcontrast enhancement, calcification, renal sinus enlargement, anterior displacement of kidneys, and other associated AML.Renal parenchymal defect, renal arterial blood supply, tumoral vessel through the renal parenchyma, dilated intratumoral vessels, intratumoral/perirenal hemorrhage, renal sinus enlargement, and associated AML were seen only or mainly in exophytic renal AML (all P value < 0.05); however, non–fat-attenuating enhancing intratumoral nodules, intratumoral calcification, and anterior displacement of the kidney were more common in liposarcoma (all P value < 0.05).AMLs reveal renal parenchymal defect at the site of tumor contact, supply from renal artery, tumoral vessel extending through the renal parenchyma, dilated intratumoral vessels, intratumoral and/or perirenal hemorrhage, renal sinus enlargement, and associated AML. Non–fat-attenuating enhancing intratumoral nodules, intratumoral calcifications, and anterior displacement of kidney were more commonly seen in liposarcoma.

Pancreatic neuroendocrine tumors containing areas of iso- or hypoattenuation in dynamic contrast-enhanced computed tomography: Spectrum of imaging findings and pathological grading

PurposeTo evaluate dynamic contrast-enhanced computed tomography (CT) features of pancreatic neuroendocrine tumors (PNETs) containing areas of iso- or hypoattenuation and the relationship with pathological grading. Materials and MethodsBetween June 2006 and March 2014, 61 PNETs in 58 consecutive patients (29 male, 29 female; median-age 55 years), which were surgically diagnosed, underwent preoperative dynamic contrast-enhanced CT. PNETs were classified based on contrast enhancement patterns in the pancreatic phase: iso/hypo-PNETs were defined as tumors containing areas of iso- or hypoattenuation except for cystic components, and hyper-PNETs were tumors showing hyperattenuation over the whole area. CT findings and contrast-enhancement patterns of the tumors were evaluated retrospectively by two radiologists and compared with the pathological grading. ResultsIso/hypo-PNETs comprised 26 tumors, and hyper-PNETs comprised 35 tumors. Not only hyper-PNETs but also most iso/hypo-PNETs showed peak enhancement in the pancreatic phase and a washout from the portal venous phase to the delayed phase. Iso/hypo-PNETs showed larger tumor size than the hyper-PNETs (mean, 3.7cm vs. 1.6cm; P<0.001), and were significantly correlated with unclear tumor margins (n=4 vs. n=0; P=0.029), the existence of cystic components (n=10 vs. n=3; P=0.006), intratumoral blood vessels in the early arterial phase (n=13 vs. n=3; P<0.001), and a smooth rim enhancement in the delayed phase (n=12 vs. n=6; P=0.019). Iso/hypo-PNETs also showed significantly higher pathological grading (WHO 2010 classification; iso/hypo, G1=14, G2=11, G3=1; hyper, G1=34, G2=1; P<0.001). ConclusionPNETs containing iso/hypo-areas showed a rapid enhancement pattern as well as hyper-PNETs, various radiological features and higher malignant potential.

Primary Pulmonary Synovial Sarcoma in a Tertiary Referral Center: Clinical Characteristics, CT, and 18F-FDG PET Findings, With Pathologic Correlations.

The purpose of this study was to describe the patient characteristics, computed tomography (CT) and F-fluorodeoxyglucose positron emission tomography (FDG PET) findings, and clinical outcomes of primary pulmonary synovial sarcoma (PPSS), together with their pathologic correlations. The medical records of 14 patients with pathologically proven PPSS in a tertiary hospital from January 1997 to December 2014 were retrospectively reviewed. The CT findings were evaluated. The maximum standardized uptake value (maxSUV) of the tumors was obtained, and clinical outcomes with respect to tumor recurrence and mortality were assessed by Kaplan-Meier analysis. The median tumor size was 10.2 cm and the most common anatomic location was the lung followed by the pleura/chest wall and mediastinum. Most of the tumors appeared as single lesions and had circumscribed margins. All the cases showed heterogeneous enhancement with necrotic or cystic portions, and intratumoral vessels were frequently seen. Half of the tumors had intratumoral calcifications, and tumor rupture, pleural/chest wall extension, and pleural effusion occurred frequently. However, lymph node enlargement was rare. The median maxSUV of the tumors was 4.35. Patient outcomes with respect to tumor recurrence (n = 8, 57.1%) and death (n = 3, 21.4%) were poor despite their young age, and the mean follow-up period was 28.5 months.In conclusion, PPSS usually occurs in young adults, generally in the lung, presents as a large, circumscribed mass, and tumor rupture or extension of the pleura/chest wall may occur. The tumors often contain calcifications and vessels; they may exhibit triple attenuation on enhanced CT images, and clinical outcomes are poor.

EGFR Regulates the Development and Microarchitecture of Intratumoral Angiogenic Vasculature Capable of Sustaining Cancer Cell Intravasation

Many malignant characteristics of cancer cells are regulated through pathways induced by the tyrosine kinase activity of the epidermal growth factor receptor (EGFR). Herein, we show that besides directly affecting the biology of cancer cells per se, EGFR also regulates the primary tumor microenvironment. Specifically, our findings demonstrate that both the expression and signaling activity of EGFR are required for the induction of a distinct intratumoral vasculature capable of sustaining tumor cell intravasation, a critical rate-limiting step in the metastatic cascade. An intravasation-sustaining mode of intratumoral angiogenic vessels depends on high levels of tumor cell EGFR and the interplay between EGFR-regulated production of interleukin 8 by tumor cells, interleukin-8–induced influx of tumor-infiltrating neutrophils delivering their unique matrix metalloproteinase-9, and neutrophil matrix metalloproteinase-9–dependent release of the vascular permeability and endothelial growth factor, VEGF. Our data indicate that through VEGF-mediated disruption of endothelial layer integrity and increase of intratumoral vasculature permeability, EGFR activity significantly facilitates active intravasation of cancer cells. Therefore, this study unraveled an important but overlooked function of EGFR in cancer, namely, its ability to create an intravasation-sustaining microenvironment within the developing primary tumor by orchestrating several interrelated processes required for the initial steps of cancer metastasis through vascular routes. Our findings also suggest that EGFR-targeted therapies might be more effective when implemented in cancer patients with early-staged primary tumors containing a VEGF-dependent angiogenic vasculature. Accordingly, early EGFR inhibition combined with various anti-VEGF approaches could synergistically suppress tumor cell intravasation through inhibiting the highly permeable angiogenic vasculature induced by EGFR-overexpressing aggressive cancer cells.

Intratumoral lymphatic vessel density as a predictor of progression‐free and overall survival in locally advanced laryngeal/hypopharyngeal cancer

Lymphatic vessel density (LVD) has been shown to be an important predictor of survival in head and neck cancers. We report the predictive value of LVD for progression-free survival (PFS) and overall survival (OS) in laryngeal/hypopharyngeal cancer. Fifty-five untreated patients with T3/T4 laryngeal and T4 hypopharyngeal cancer underwent laryngectomy between 1999 and 2010. Surgical specimens were immunostained with D2-40, a specific lymphatic marker. LVDs were determined in tumor vessel "hot spots." Recursive partitioning analysis identified LVD thresholds for both peritumoral (LVDpt) and intratumoral (LVDit) vessels for association with PFS and OS. Patients with mean LVDit of <11 vessels/mm(2) had 2-year PFS and OS rates of 58% and 65%, respectively, compared to 13% and 13% for those with LVDit ≥11 vessels/mm(2) (p = .06 and .04, respectively). Intratumoral lymphatic vessel density is predictive of PFS and OS in locally advanced laryngeal/hypopharyngeal cancer. © 2015 Wiley Periodicals, Inc. Head Neck 38: E417-E420, 2016.

CT imaging of ovarian yolk sac tumor with emphasis on differential diagnosis.

Ovarian yolk sac tumors (YSTs) are rare neoplasms. No radiological study has been done to compare the imaging findings between this type of tumor and other ovarian tumors. Here we analyzed the CT findings of 11 pathologically proven ovarian YSTs and compared their imaging findings with 18 other types of ovarian tumors in the same age range. Patient age, tumor size, tumor shape, ascites and metastasis of two groups did not differ significantly (P > 0.05). A mixed solid-cystic nature, intratumoral hemorrhage, marked enhancement and dilated intratumoral vessel of two groups differed significantly (P < 0.05). The area under the ROC curve of four significant CT features was 0.679, 0.707, 0.705, and 1.000, respectively. Multivariate logistic regression analysis identified two independent signs of YST: intratumoral hemorrhage and marked enhancement. Our results show that certain suggestive CT signs that may be valuable for improving the accuracy of imaging diagnosis of YST and may be helpful in distinguishing YST from other ovarian tumors.

Stress-mediated progression of solid tumors: effect of mechanical stress on tissue oxygenation, cancer cell proliferation, and drug delivery.

Oxygen supply plays a central role in cancer cell proliferation. While vascular density increases at the early stages of carcinogenesis, mechanical solid stresses developed during growth compress tumor blood vessels and, thus, drastically reduce not only the supply of oxygen, but also the delivery of drugs at inner tumor regions. Among other effects, hypoxia and reduced drug delivery compromise the efficacy of radiation and chemo/nanotherapy, respectively. In the present study, we developed a mathematical model of tumor growth to investigate the interconnections among tumor oxygenation that supports cancer cell proliferation, the heterogeneous accumulation of mechanical stresses owing to tumor growth, the non-uniform compression of intratumoral blood vessels due to the mechanical stresses, and the insufficient delivery of oxygen and therapeutic agents because of vessel compression. We found that the high vascular density and increased cancer cell proliferation often observed in the periphery compared to the interior of a tumor can be attributed to heterogeneous solid stress accumulation. Highly vascularized peripheral regions are also associated with greater oxygenation compared with the compressed, less vascularized inner regions. We also modeled the delivery of drugs of two distinct sizes, namely chemotherapy and nanomedicine. Model predictions suggest that drug delivery is affected negatively by vessel compression independently of the size of the therapeutic agent. Finally, we demonstrated the applicability of our model to actual geometries, employing a breast tumor model derived from MR images.

CT Characterization of Duodenal Gastrointestinal Stromal Tumors.

Diagnosis and staging of duodenal gastrointestinal stromal tumors (GISTs) by noninvasive imaging is critical for effective treatment, but the imaging features of duodenal GISTs remain largely undefined because of their rarity. The purpose of this article was to characterize duodenal GISTs using CT. Thirty-four patients with duodenal GISTs were analyzed by clinical symptom evaluation, pathologic examination, and CT in this retrospective study. Unenhanced and contrast-enhanced examinations were performed in all patients. Imaging characteristics, including the lesion location, size, growth pattern, ulceration, internal components, arterial blood supply, intratumoral arterioportal shunting, intratumoral vessels, rim enhancement, and enhancement patterns were reviewed. The duodenal GISTs were solitary masses with well-defined margins. The average diameter was 7.1 cm. The second portion was the most common site (20/34). Ulceration was a common feature (15/34). Calcification was uncommon (3/34), and mixed growth pattern was more common (26/34). Rim enhancement (24/34) and mixed enhancement pattern (15/34) were common. Arterial blood supply, intratumoral vasculature, and draining veins were all detected and were obvious on the arterial phase. The portal venous trunk and superior mesenteric vein were the main veins into which early arterioportal shunting drained. Primary duodenal GISTs are generally large, well-defined, heterogeneously enhancing, and hypervascular masses with a prominent mixed growth pattern on CT images. Our findings suggest that CT can help depict the origin of the tumoral arteries and draining veins on the arterial phase and may be a key defining diagnostic feature for duodenal GISTs.

Significance of Vascular Endothelial Growth Factor (VEGF)-C and VEGF-D in the Progression of Cutaneous Melanoma.

Induction of tumor lymphangiogenesis by vascular endothelial growth factor (VEGF)-C and VEGF-D promotes metastasis in many human cancers. The aim of this study was to examine the role of VEGF-C and VEGF-D in lymphangiogenesis and lymph node metastasis in patients with cutaneous melanoma. Fifty-four melanoma specimens (18 with lymph node metastasis, 36 nonmetastatic) were investigated by immunostaining for VEGF-C, VEGF-D, and for lymphatic endothelial marker D2-40. VEGF-C and VEGF-D expression was assessed as a percentage and intensity of stained tumor cells, tumor-associated macrophages and fibroblasts. The quantification of lymphangiogenesis was conducted by computer-assisted morphometric analysis. The expressions of both VEGF-C and VEGF-D in tumor cells were significantly higher in lymph node metastatic melanomas compared with nonmetastatic melanomas (P = .015 VEGF-C; P = .005 VEGF-D). There was no statistically significant difference between metastatic and nonmetastatic melanomas regarding the expression of VEGF-C and VEGF-D in macrophages and fibroblasts. Metastatic melanomas showed a significantly higher intratumoral and peritumoral lymphatic vessel density (LVD) compared with nonmetastatic melanomas (P = .000 intratumoral, P = .000 peritumoral). Melanomas with VEGF-C positive tumor cells showed a significantly higher intratumoral and peritumoral LVD compared with VEGF-C negative tumor cells group of melanomas (P = .006 intratumoral, P = .010 peritumoral). VEGF-C expression in macrophages, fibroblasts, as well as VEGF-D expression in tumor cells, macrophages, and fibroblasts, showed no correlation with the intratumoral and peritumoral LVD. Our findings show the significance of VEGF-C in tumor cells in the induction of intratumoral and peritumoral lymphangiogenesis. This study suggests that both VEGF-C and VEGF-D in tumor cells promote lymph node metastasis, and that the immunohistochemical analysis of expression can be a useful tool for predicting clinical behavior of cutaneous melanoma.

IKKβ Regulates VEGF Expression and Is a Potential Therapeutic Target for Ovarian Cancer as an Antiangiogenic Treatment.

The prolongation of progression-free survival (PFS) in patients with advanced ovarian cancer by antiangiogenic therapy has been shown in several clinical trials. However, although an anti-VEGF antibody (bevacizumab) is the only option currently available, its efficacy is limited and it is not cost effective for use in all patients. Therefore, the development of a novel antiangiogenic drug, especially composed of small-molecule compounds, could be a powerful armament for ovarian cancer treatment. As NF-κB signaling has the potential to regulate VEGF expression, we determined to identify whether VEGF expression is associated with NF-κB activation and to investigate the possibility of a novel IKKβ inhibitor, IMD-0354 (IMMD Inc.), as an antiangiogenic drug. Tissue microarrays from 94 ovarian cancer tissues were constructed and immunohistochemical analyses performed. We revealed that IKK phosphorylation is an independent prognostic factor (PFS: 26.1 vs. 49.8 months, P = 0.011), and is positively correlated with high VEGF expression. In in vitro analyses, IMD-0354 robustly inhibited adhesive and invasive activities of ovarian cancer cells without impairing cell viabilities. IMD-0354 significantly suppressed VEGF production from cancer cells, which led to the inhibition of angiogenesis. In a xenograft model, the treatment of IMD-0354 significantly inhibited peritoneal dissemination with a marked reduction of intratumoral blood vessel formation followed by the inhibition of VEGF expression from cancer cells. IMD-0354 is a stable small-molecule drug and has already been administered safely to humans in other trials. Antiangiogenic therapy targeting IKKβ is a potential future option to treat ovarian cancer.