Function Of Tumor Vasculature Research Articles

Normalization of Tumor Vasculature by Oxygen Microbubbles with Ultrasound.

Tumor microenvironment influences the efficacy of anti-cancer therapies. The dysfunctional tumor vasculature limits the efficiency of oxygenation and drug delivery to reduce treatment outcome. A concept of tumor vascular normalization (VN), which inhibits angiogenesis to improve vessel maturity, blood perfusion, and oxygenation, has been demonstrated under the anti-angiogenic therapy. The efficiency of drug delivery and penetration is increased by enhancing perfusion and reducing interstitial fluid pressure during the time window of VN. However, anti-angiogenic agents only induce transient VN and then prune vessels to aggravate tumor hypoxia. To repair tumor vessels without altering vessel density, we proposed to induce tumor VN by local oxygen release via oxygen microbubbles with ultrasound. With tumor perfusion enhancement under ultrasound contrast imaging tracing, the time window of VN was defined as 2-8 days after a single oxygen microbubble treatment. The enhanced tumor oxygenation after oxygen microbubble treatment inhibited hypoxia inducible factor-1 alpha (HIF-1α)/vascular endothelial growth factor (VEGF) pathway to improve the morphology and function of tumor vasculature. The pericyte coverage and Hoechst penetration of tumor vessels increased without any changes to the vessel density. Finally, the intratumoral accumulation of anti-cancer drug doxorubicin could be increased 3-4 folds during tumor VN. These findings demonstrate that regulating tumor oxygenation by oxygen microbubbles could normalize dysfunctional vessels to enhance vascular maturity, blood perfusion, and drug penetration. Furthermore, ultrasound perfusion imaging provides a simple and non-invasive way to detect the VN time window, which increases the feasibility of VN in clinical cancer applications.

Tanshinone II A improves distribution and anti-tumor efficacy of pegylated liposomal doxorubicin via normalizing the structure and function of tumor vasculature in hepa1-6 hepatoma mice model

OBJECTIVETo investigate whether the Tan II A could improve the distribution and anti-tumor efficacy of Pegylated Liposomal Doxorubicin (PLD) via normalizing the structure and function of vasculature in Hepa1-6 hepatoma mice model. METHODSHepa1-6 hepatoma-bearing mice were treated with Tan II A for 14 d. Distribution and anti-tumor efficacy of PLD, and the structure and function of the tumor vasculature were evaluated using various techniques. RESULTSTan II A significantly reduced the micro-vessel density (MVD). After Tan II A treatment, the tumor vascular walls were better structured, as the increased coverage of the pericytes and the promoted contact of the basement membrane and endothelial cell. Functional tests showed that tumor hypoxia was improved and the exudation amount of Evans blue in the parenchyma of the tumor decreased. In addition, mice treated with Tan II A had greater PLD penetration distance intratumorally. Furthermore, combined therapy of Tan II A and PLD significantly inhibited tumor growth. CONCLUSIONThis study suggests that Tan II A helps normalizing the tumor vasculature and has therapeutic potential in increasing the distribution of chemotherapy drug in the tumor.

Abstract B14: The effect of sunitinib treatment in human melanoma xenografts: Associations with angiogenic profiles

Abstract The effect of antiangiogenic agents targeting the vascular endothelial growth factor A (VEGF-A) pathway has been reported to vary substantially in preclinical studies. Treatments with these agents have induced substantial reductions in vessel density in some tumor models, and minor or no reductions in vessel density in others. Additionally, the treatments have improved vascular function and oxygenation in some tumor models, and induced hypoxia in others. The purpose of this study was to investigate the effect of sunitinib treatment on the morphology and function of tumor vasculature in melanoma xenografts with different angiogenic profiles. A-07, U-25, D-12, and R-18 melanoma xenografts grown in dorsal window chambers were used as preclinical tumor models. Tumor-bearing mice were treated with sunitinib (40 mg/kg/day) or vehicle. Morphologic parameters of tumor vascular networks were assessed from high-resolution transillumination images, and tumor blood supply times were assessed from first-pass imaging movies recorded after a bolus of 155 kD tetramethylrhodamine isothiocyanate-labeled dextran was injected intravenously. Tumor hypoxia was assessed in immunohistochemical preparations by using pimonidazole as hypoxia marker, and the gene expression and the protein secretion rate of angiogenic factors were assessed by quantitative PCR and ELISA, respectively. The melanoma lines differed substantially in the expression of VEGF-A, VEGF-C, and platelet-derived growth factor A. Sunitinib treatment reduced vessel densities and induced hypoxia in all melanoma lines, and the magnitude of the effect was associated with the gene expression and protein secretion rate of VEGF-A. Sunitinib treatment also increased vessel segment lengths, reduced the number of small-diameter vessels, and inhibited growth-induced increases in the diameter of surviving vessels, but did not affect blood supply times. In conclusion, sunitinib treatment did not improve vascular function but reduced vessel density and induced hypoxia in human melanoma xenografts. The magnitude of the treatment-induced effect was associated with the angiogenic profiles of the melanoma lines. Citation Format: Jon-Vidar Gaustad, Trude G. Simonsen, Lise Mari K. Andersen, Einar K. Rofstad. The effect of sunitinib treatment in human melanoma xenografts: Associations with angiogenic profiles. [abstract]. In: Proceedings of the AACR Special Conference on Engineering and Physical Sciences in Oncology; 2016 Jun 25-28; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2017;77(2 Suppl):Abstract nr B14.

Abstract A41: Use of patient-derived tumor xenografts (PDX) for discovery and development of an anti-Notch2/3 monoclonal antibody targeting cancer stem cells

Abstract Accumulating evidence has suggested that tumors may arise and grow as a result of the formation of a subset cell population termed cancer stem cells (CSC) or tumor initiating cells. Several research reports have indicated that CSCs are relatively resistant to conventional therapies. Thus, therapeutic strategies that specifically target cancer stem cells could have a major impact on cancer patient survival. Patient-derived tumor xenografts (PDX) have played a major role in the development of new cancer therapies. The advantage of PDX over standard cell-line xenograft models is that PDX retain much of the molecular, genetic, and histological heterogeneity of the original tumor and are minimally passed. In addition, self-renewal and lineage differentiation, the hallmarks of cancer stem cells, can be demonstrated through serial transplantation in immunodeficient mice. Our therapeutic approach in cancer stem cell drug discovery has been to target key developmental pathways that have been strongly implicated in cancer including the Notch pathway. We carried out tumorigenicity studies in three SCLC PDX models utilizing naïve cells positively enriched for either Notch2 or Notch3 expression. In two of the three models, Notch2+ and/or Notch3+ cell populations resulted in enhanced tumor formation. In a SCLC tumor that expressed low levels of Notch3, the Notch2+ population resulted in 70% tumor formation compared to CD44+ (10%) or CD133+ (45%) enriched populations. We have developed a monoclonal antibody, tarextumab (OMP-59R5), which selectively inhibits the function of both Notch2 and Notch3. Our preclinical data in PDX models demonstrate that tarextumab was efficacious in inhibiting the growth of various indications with minimal intestinal toxicity. Notably, the sensitivity of tarextumab in combination with gemcitabine or gemcitabine plus nab-paclitaxel in pancreatic tumors was associated with higher levels of Notch3 gene expression. Interference with Notch2/3 signaling by tarextumab delays tumor recurrence, decreases cancer stem cell frequency (as determined by in vivo LDA studies) and modulates the function of tumor vasculature. Our ALPINE Phase 1b clinical trial indicates that tarextumab is generally well-tolerated and shows signs of anti-tumor efficacy and modulation of Notch pathway signaling in the clinic. Furthermore, we observe a higher response rate and longer survival in patients with Notch3 high tumors receiving gemcitabine/nab-paclitaxel/tarextumab combination therapy. Analysis of pre- and post-treatment tumor biopsies showed an inhibition of Notch pathway and CSC gene signatures. Ongoing Phase 2 clinical trials evaluate first-line treatment with tarextumab in metastatic pancreatic cancer (ALPINE) and small cell lung cancer (PINNACLE). Collectively, these results demonstrate the utility of PDX models for discovery and development of anti-cancer stem cell therapeutics, identifying pharmacodynamic endpoints of drug actions, identifying predictive biomarkers for patient stratification, and translating these preclinical findings into clinical trials. Citation Format: Marcus M. Fischer, Wan-Ching Yen, Fumiko Axelrod, Christopher Bond, Jennifer Cain, Belinda Cancilla, Randall Henner, Rene Meisner, Aaron Sato, Jalpa Shaw, Tracy Tang, Breanna Wallace, Min Wang, Chun Zhang, Ann Kapoun, Lei Zhou, Jakob Dupont, John Lewicki, Austin Gurney, Tim Hoey. Use of patient-derived tumor xenografts (PDX) for discovery and development of an anti-Notch2/3 monoclonal antibody targeting cancer stem cells. [abstract]. In: Proceedings of the AACR Special Conference: Patient-Derived Cancer Models: Present and Future Applications from Basic Science to the Clinic; Feb 11-14, 2016; New Orleans, LA. Philadelphia (PA): AACR; Clin Cancer Res 2016;22(16_Suppl):Abstract nr A41.

Abstract A03: Vascular function affected fundamentally different by different antiangiogenic agents

Abstract The purpose of this preclinical study was to investigate the effect of two antiangiogenic agents, properdistatin and sunitinib, on the morphology and function of tumor vasculature. Properdistatin is a novel peptide derived from the thrombospondin-1 domain of the plasma protein properdin. Properdistatin treatment has been shown to inhibit angiogenesis and tumor growth in breast cancer xenografts. Sunitinib is a tyrosine kinase inhibitor that interacts with several receptors including vascular endothelial growth factor receptors 1-3, and platelet-derived growth factor receptors α-β. Sunitinib treatment has been shown to inhibit angiogenesis and tumor growth in a number of tumor models, and has been approved clinically for several indications. A-07 human melanoma xenografts grown in dorsal window chambers or as intradermal flank tumors were used as preclinical tumor model. In window chamber tumors, morphologic parameters of tumor vascular networks were assessed from high-resolution transillumination images, and blood supply time and plasma velocities were assessed from first-pass imaging movies recorded after a bolus of 155kDa tetramethylrhodamine isothiocyanate-labeled dextran had been administered intravenously. In flank tumors, parametric images of ADC and Ktrans were produced from diffusion weighted magnetic resonance imaging (DW-MRI) and dynamic contrast-enhanced MRI (DCE-MRI). Tumor hypoxia and necrosis were assessed from immunohistochemical preparations of the imaged tissue, and tumor interstitial fluid pressure (IFP) was assessed with probe measurement. Properdistatin treatment reduced the density of small-diameter vessels, reduced blood supply time, and increased plasma velocities, whereas sunitinib treatment did not alter blood supply time, reduced overall vessel density, induced tumor hypoxia and necrosis, and did not alter tumor IFP. Sunitinib-treated flank tumors showed reduced Ktrans values corresponding to a decrease in vessel density, and increased ADC values corresponding to induction of tumor necrosis. In conclusion, properdistatin treatment improved vascular function, whereas sunitinib treatment did not improve vascular function but induced tumor hypoxia and necrosis in A-07 tumors. This data demonstrates that different antiangiogenic agents may affect vascular function fundamentally different in the same tumor model. DW-MRI and DCE-MRI were sensitive to the sunitinib-induced changes in the tumor microenvironment, demonstrating that these MR-techniques may be used to identify tumors where antiangiogenic treatment does not improve vascular function. Citation Format: Jon-Vidar Gaustad, Trude G. Simonsen, Tord Hompland, Lise Mari K. Andersen, Einar K. Rofstad. Vascular function affected fundamentally different by different antiangiogenic agents. [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 A03.

Thrombospondin-1 domain-containing peptide properdistatin improves vascular function in human melanoma xenografts

Properdistatin is a novel peptide derived from the thrombospondin-1 domain of the plasma protein properdin. The purpose of this study was to investigate the effect of properdistatin treatment on the morphology and function of tumor vasculature. A-07 human melanoma xenografts grown in dorsal window chambers were used as preclinical model. Tumors were treated with 80mg/kg/day properdistatin or vehicle for 4days. Morphologic parameters of tumor vascular networks were assessed from high-resolution transillumination images, and tumor blood supply time and plasma velocities were assessed from first-pass imaging movies recorded after a bolus of 155kDa tetramethylrhodamine isothiocyanate-labeled dextran had been administered intravenously. Properdistatin-treated tumors showed reduced density of small-diameter vessels, reduced blood supply time, and increased plasma velocities. In conclusion, properdistatin treatment inhibited angiogenesis and improved vascular function in A-07 tumors.

Polarization of tumor-associated macrophage is associated with tumor vascular normalization by endostatin.

Vascular normalization is an emerging concept in cancer treatment, but its precise mechanisms are not completely understood. The polarization of tumor-associated macrophages (TAMs) is important in tumor angiogenesis and metastasis. However, little is known about the effect of anti-angiogenic agents on the polarization of tumor-associated macrophages. Therefore, we explore the changes of TAMs polarization in the development of tumor vascular normalization induced by endostatin. A murine xenograft model of lung cancer was treated with endostatin for 10 days. The morphology and function of tumor vasculature was examined using various techniques. Flow cytometry was carried out to assess the TAMs, and immunofluorescence was used to examine Tie-2-expressing monocytes (TEMs) in tumors. Levels of the histidine-rich glycoprotein (HRG) in tumors were measured by immunohistochemistry and Western blot. Tumor vessels became more normal and mature on day six in the endostatin-treated mice. During vascular normalization, the number of M2-like TAMs and TEMs in the tumors was significantly reduced, whereas the number of M1-like TAMs showed an increase on day six after endostatin treatment, although the latter was not statistically significant. The HRG in the tumors accumulated at an early stage after endostatin administration. The polarization of TAMs is associated with tumor vascular normalization induced by endostatin. These observations may be useful in the exploration of new strategies for anti-angiogenic treatment.

Sunitinib treatment does not improve blood supply but induces hypoxia in human melanoma xenografts

BackgroundAntiangiogenic agents that disrupt the vascular endothelial growth factor pathway have been demonstrated to normalize tumor vasculature and improve tumor oxygenation in some studies and to induce hypoxia in others. The aim of this preclinical study was to investigate the effect of sunitinib treatment on the morphology and function of tumor vasculature and on tumor oxygenation.MethodsA-07-GFP and R-18-GFP human melanoma xenografts grown in dorsal window chambers were used as preclinical tumor models. Morphologic parameters of tumor vascular networks were assessed from high-resolution transillumination images, and tumor blood supply time was assessed from first-pass imaging movies recorded after a bolus of 155 kDa tetramethylrhodamine isothiocyanate-labeled dextran had been administered intravenously. Tumor hypoxia was assessed from immunohistochemical preparations of the imaged tissue by use of pimonidazole as a hypoxia marker.ResultsSunitinib treatment reduced vessel densities, increased vessel segment lengths, did not affect blood supply times, and increased hypoxic area fractions.ConclusionSunitinib treatment did not improve vascular function but induced hypoxia in A-07-GFP and R-18-GFP tumors.

Low-dose endostatin normalizes the structure and function of tumor vasculature and improves the delivery and anti-tumor efficacy of cytotoxic drugs in a lung cancer xenograft murine model.

To some extent endostatin normalizes tumor vasculature. However, the optimum time window and optimum drug dose for tumor vascular normalization need to be explored. Here we investigate the effect of low-dose endostatin on the structure and function of tumor vasculature and the delivery and anti-tumor efficacy of cytotoxic drugs. A lung cancer xenograft murine model was treated with low-dose endostatin for 10 days. The structure and function of the tumor vasculature were evaluated using various techniques. Paclitaxel was added in different schedules. Endostatin caused a significant reduction in microvessel density. Tumor vascular walls after endostatin treatment were better structured. Tumor blood perfusion was increased on day six after endostatin administration. On days three, six, and 10, Evans blue extravasation into the parenchyma of tumors was decreased. On days three and six, endostatin-treated mice had greater paclitaxel delivery. The time window of vascular normalization was approximately three to six days. On days one to three, and days four to six, combined therapy with paclitaxel significantly inhibited tumor growth. Low-dose endostatin aids normalization of tumor vasculature. This resulted in improved delivery of cytotoxic drugs to the tumor, which closely correlates with synergistic efficacy when combined with paclitaxel during the normalization window.

Irinophore C, a Novel Nanoformulation of Irinotecan, Alters Tumor Vascular Function and Enhances the Distribution of 5-Fluorouracil and Doxorubicin

To examine the antitumor effects of Irinophore C, a nanopharmaceutical formulation of irinotecan, on the tissue morphology and function of tumor vasculature in HT-29 human colorectal tumors. Fluorescence microscopy was used to map and quantify changes in tissue density, tumor vasculature, hypoxia, and the distribution of Hoechst 33342, a perfusion marker, and the anticancer drug, doxorubicin. Noninvasive magnetic resonance imaging was used to quantify Ktrans, the volume transfer constant of a solute between the blood vessels and extracellular tissue compartment of the tumor, as a measure of vascular function. Following treatment with Irinophore C, 19F magnetic resonance spectroscopy was used to monitor the delivery of 5-fluorouracil (5-FU) to the tumor tissue, whereas scintigraphy was used to quantify the presence of bound [14C]5-FU. Irinophore C decreased cell density (P = 8.42 x 10(-5)), the overall number of endothelial cells in the entire section (P = 0.014), tumor hypoxia (P = 5.32 x 10(-9)), and K(trans) (P = 0.050). However, treatment increased the ratio of endothelial cells to cell density (P = 0.00024) and the accumulation of Hoechst 33342 (P = 0.022), doxorubicin (P = 0.243 x 10(-5)), and 5-FU (P = 0.0002) in the tumor. Vascular endothelial growth factor and interleukin-8, two proangiogenic factors, were down-regulated, whereas the antiangiogenic factor TIMP-1 was up-regulated in Irinophore C-treated tumors. Irinophore C treatment improves the vascular function of the tumor, thereby reducing tumor hypoxia and increasing the delivery and accumulation of a second drug. Reducing hypoxia would enhance radiotherapy, whereas improving delivery of a second drug to the tumor should result in higher cell kill.