Tumor Vascular Permeability Research Articles

Focal Adhesion Kinase (FAK) tyrosine 397E mutation restores the vascular leakage defect in endothelium‐specific FAK‐kinase dead mice

Focal adhesion kinase (FAK) inhibitors have been developed as potential anticancer agents and are undergoing clinical trials. In vitro activation of the FAK kinase domain triggers autophosphorylation of Y397, Src activation, and subsequent phosphorylation of other FAK tyrosine residues. However, how FAK Y397 mutations affect FAK kinase‐dead (KD) phenotypes in tumour angiogenesis in vivo is unknown. We developed three Pdgfb‐iCreert‐driven endothelial cell (EC)‐specific, tamoxifen‐inducible homozygous mutant mouse lines: FAK wild‐type (WT), FAK KD, and FAK double mutant (DM), i.e. KD with a putatively phosphomimetic Y397E mutation. These ECCre+;FAKWT / WT, ECCre+;FAKKD / KD and ECCre+;FAKDM / DM mice were injected subcutaneously with syngeneic B16F0 melanoma cells. Tumour growth and tumour blood vessel functions were unchanged between ECCre+;FAKWT / WT and ECCre−;FAKWT / WT control mice. In contrast, tumour growth and vessel density were decreased in ECCre+;FAKKD / KD and ECCre+;FAKDM / DM mice, as compared with Cre − littermates. Despite no change in the percentage of perfused vessels or pericyte coverage in either genotype, tumour hypoxia was elevated in ECCre+;FAKKD / KD and ECCre+;FAKDM / DM mice. Furthermore, although ECCre+;FAKKD / KD mice showed reduced blood vessel leakage, ECCre+;FAKDM / DM and ECCre−;FAKDM / DM mice showed no difference in leakage. Mechanistically, fibronectin‐stimulated Y397 autophosphorylation was reduced in Cre+;FAKKD / KD ECs as compared with Cre+;FAKWT / WT cells, with no change in phosphorylation of the known Src targets FAK‐Y577, FAK‐Y861, FAK‐Y925, paxillin‐Y118, p130Cas‐Y410. Cre+;FAKDM / DM ECs showed decreased Src target phosphorylation levels, suggesting that the Y397E substitution actually disrupted Src activation. Reduced VE‐cadherin‐pY658 levels in Cre+;FAKKD / KD ECs were rescued in Cre+FAKDM / DM ECs, corresponding with the rescue in vessel leakage in the ECCre+;FAKDM / DM mice. We show that EC‐specific FAK kinase activity is required for tumour growth, angiogenesis, and vascular permeability. The ECCre+;FAKDM/DM mice restored the KD‐dependent tumour vascular leakage observed in ECCre+;FAKKD/KD mice in vivo. This study opens new fields in in vivo FAK signalling. © 2017 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.

Xuebijing Injection (血必净注射液) increases early survival rate by alleviating pulmonary vasopermeability in rats subjected to severe burns

To investigate the effects of Xuebijing Injection (, XBJ) on survival rate and pulmonary vasopermeability in a rat model of severe scald injury. Rats were divided into two experiments: experiment 1 was monitored for 12 h post-injury for survival analysis after severe burns; in experiment 2, rats were killed for determination of pulmonary vascular permeability and pro-inflflammatory mediators. In both experiments, rats were subject to third-degree 50% total body surface area (TBSA) burns or sham injury followed by XBJ or normal saline (NS) treatment. In addition, rat pulmonary microvascular endothelium cells (PMECs) were pretreated with either XBJ or phosphate buffer saline (PBS), and then subjected to sham serum or scald serum stimulation for 2 or 6 h, followed by transwell examination for the permeability of PMECs. Meanwhile, pro-inflflammatory mediators in PMECs culture supernatant were also investigated. The average survival time in the scald+XBJ group was 582.1±21.2 min, which was signifificantly longer than that in the scald + NS group (345.8±25.4 min, P<0.01). Plasma levels of tumor necrosis factor-alpha (TNF-α), E-selectin, interleukin-6 (IL-6), vascular permeability and water content of lung tissues were signifificantly increased in animals after severe burns (P<0.01). However, administration of XBJ signifificantly decreased these levels in plasma and lung tissue. In in vitro cell experiments, XBJ markedly attenuated permeability in PMECs monolayer and reduced the levels of TNF-α, IL-6 and soluble E-selectin after stimulation with scald serum (P<0.01). XBJ increases early survival rate by alleviating pulmonary vasopermeability and inhibiting pro-inflflammatory mediators in rats subjected to lethal scald injury. XBJ may be a potent drug in treatment of severe burns.

Permeability of Brain Tumor Vessels Induced by Uniform or Spatially Microfractionated Synchrotron Radiation Therapies

Tocompare the blood-brain barrier permeability changes induced by synchrotron microbeam radiation therapy (MRT, which relies on spatial fractionation of the incident x-ray beam into parallel micron-wide beams) with changes induced by a spatially uniform synchrotron x-ray radiation therapy. Male rats bearing malignant intracranial F98 gliomas were randomized into 3 groups: untreated, exposed to MRT (peak and valley dose: 241 and 10.5Gy, respectively), or exposed to broad beam irradiation (BB) delivered at comparable doses (ie, equivalent to MRT valley dose); both applied by 2 arrays, intersecting orthogonally the tumor region. Vessel permeability was monitored invivo by magnetic resonance imaging 1day before (T-1) and 1, 2, 7, and 14days after treatment start. To determine whether physiologic parameters influence vascular permeability, we evaluated vessel integrity in the tumor area with different values for cerebral blood flow, blood volume, edema, and tissue oxygenation. Microbeam radiation therapy does not modify the vascular permeability of normal brain tissue. Microbeam radiation therapy-induced increase of tumor vascular permeability was detectable from T2 with a maximum at T7 after exposure, whereas BB enhanced vessel permeability only at T7. At this stage MRT was more efficient at increasing tumor vessel permeability (BB vs untreated: +19.1%; P=.0467; MRT vs untreated: +44.8%; P<.0001), and its effects lasted until T14 (MRT vs BB, +22.6%; P=.0199). We also showed that MRT was more efficient at targeting highly oxygenated (high blood volume and flow) and more proliferative parts of the tumor than BB. Microbeam radiation therapy-induced increased tumor vascular permeability is: (1) significantly greater; (2) earlier and more prolonged than that induced by BB irradiation, especially in highly proliferative tumor areas; and (3) targets all tumor areas discriminated by physiologic characteristics, including those not damaged by homogeneous irradiation.

Quantification of Tumor Vascular Permeability and Blood Volume by Positron Emission Tomography.

Purpose: Evans Blue (EB) is an azo dye that binds quantitatively with serum albumin. With an albumin binding, NOTA conjugated truncated Evan's blue (NEB) dye derived PET tracer, we aimed to establish a strategy for evaluating vascular permeability in malignant tumors via non-invasive PET. Experimental design: Sixty-minute dynamic PET using [18F]FAl-NEB was performed in three xenograft tumor models including INS-1 rat insulinoma, UM-SCC-22B human head and neck carcinoma and U-87 MG human glioblastoma. Tumor vascular permeability was quantified by the difference of the slopes between tumor and blood time-activity curve (TACs, expressed as Ps). The method was further substantiated by EB extraction and colorimetric assay and correlates with that calculated from dynamic contrast enhanced magnetic resonance imaging (DCE-MRI). The changes in tumor vasculature at different time points were assessed with NEB PET in U-87 MG and UM-SCC-22B tumor models after treatment with bevacizumab or doxorubicin. Result: The Ps values calculated from tumor and blood TACs from multiple time-point static images are consistent with those from dynamic images. Moreover, the Ps showed a positive and significant correlation with extracted EB concentration and KPS-MRI generated from DCE-MRI, which further confirmed the soundness of this methodology. The antiangiogenic effect of bevacizumab could be revealed by NEB PET in U-87 MG tumors as early as 8 hrs after therapy, demonstrated by a substantial decrease of Ps. On the contrary, there was no significant change of Ps in bevacizumab treated UM-SCC-22B tumors, compared with control group. However, the significant changes of Pswere overestimated in doxorubicin treated UM-SCC-22B tumors. Conclusions: We successfully developed a relatively convenient and novel strategy to evaluate vascular permeability and blood volume using NEB PET. This method will be advantageous in evaluating vascular permeability, promoting drug delivery, and monitoring tumor response to therapeutics that affect tumor angiogenesis.

Highly sensitive detection of invasive lung cancer cells by novel antibody against amino-terminal domain of laminin γ2 chain.

The laminin γ2 chain, a subunit of laminin‐332 (α3β3γ2), is a molecular marker for invasive cancer cells, but its pathological roles in tumor progression remain to be clarified. It was recently found that the most N‐terminal, domain V (dV) of γ2 chain has activities to bind CD44 and stimulate tumor cell migration and vascular permeability. In the present study, we prepared a mAb recognizing γ2 dV. Immunoblotting with this antibody, for the first time, showed that proteolytic fragments containing dV in a range of 15–80 kDa were highly produced in various human cancer cell lines and lung cancer tissues. In immunohistochemistry of adenocarcinomas and squamous cell carcinomas of the lung, this antibody immunostained the cytoplasm of invasive tumor cells and adjacent stroma much more strongly than a widely used antibody recognizing the C‐terminal core part of the processed γ2 chain. This suggests that the dV fragments are highly accumulated in tumor cells and stroma compared to the processed γ2 protein. The strong tumor cell staining with the dV antibody correlated with the tumor malignancy grade. We also found that the laminin β3 and α3 chains were frequently overexpressed in tumor cells and tumor stroma, respectively. The cytoplasmic dV detection was especially prominent in tumor cells infiltrating stroma, but low in the cells surrounded by basement membranes, suggesting that the active tumor–stroma interaction is critical for the aberrant γ2 expression. The present study suggests important roles of laminin γ2 N‐terminal fragments in tumor progression.

NIMG-22. MRI CHANGES IN NEWLY DIAGNOSED GLIOBLASTOMA DURING CHEMORADIATION AND ADJUVANT TEMOZOLOMIDE

Characterize anatomic and functional MRI changes in patients with newly diagnosed glioblastoma (nGBM) to more accurately measure impact of treatment. The standard of care for nGBM is maximal safe resection, six weeks of concurrent chemoradiation (CRT), followed by adjuvant monthly temozolomide. FLAIR and T1 post-contrast sequences are frequently used to assess treatment response but poorly reflect underlying tumor behavior with respect to tumor cellularity, vascular permeability, and perfusion. Following IRB approval, patients with nGBM underwent MR imaging before treatment, during week 6 of CRT, four weeks post-CRT, and before cycle 2 of adjuvant temozolomide (“pre-cycle 2”). We measured FLAIR and contrast-enhancing (CE) volumes, apparent diffusion coefficient (ADC; reflecting tumor cellularity), Ktrans (reflecting vascular permeability), and relative cerebral blood volume (rCBV) and flow (rCBF; reflecting perfusion) on spin echo (SE) and gradient echo (GE) sequences (rCBVSE, rCBFSE, rCBVGE, rCBFGE, respectively). Fourteen patients were included. Percent changes during treatment were compared to pre-treatment values. Within the CE region of interest (ROI), the respective median percent changes during CRT, post-CRT, and pre-cycle 2 were: CE volume -0.75%, 19.98%, 5.93%; ADC 10.15%, 18.03%, 27.21%; Ktrans 32.05%, 22.48%, 7.06%; rCBVSE -18.82%, -33.21%, -32.86%; rCBFSE -11.42%, -28.80%, -26.99%; rCBVGE -17.05%, -23.92%, -30.39%; and rCBFGE -21.98%, -24.29%, -32.12%. Within the peri-tumoral FLAIR ROI, the respective median percent changes during CRT, post-CRT, and pre-cycle 2 were: FLAIR volume 20.36%, 80.29%, 69.36%; ADC -3.63%, 6.59%, 11.51%; Ktrans 5.88%, -7.40%, -31.45%; rCBVSE -11.62%, -34.60%, -20.18%; rCBFSE -0.31%, -22.42%, -25.23%; rCBVGE -0.80%, -13.05%, -22.32%; and rCBFGE 6.30%, -6.25%, -19.57%. Chemoradiation in nGBM results in increased edema and permeability early during treatment, likely related to tumoral and peri-tumoral inflammation; decreased tumor cellularity, reflected by increased ADC; and decreased perfusion. ADC, Ktrans, and perfusion parameters are thus helpful in better characterizing tumor microenvironment during treatment.

ACRIN 6684: Assessment of Tumor Hypoxia in Newly Diagnosed Glioblastoma Using 18F-FMISO PET and MRI.

Structural and functional alterations in tumor vasculature are thought to contribute to tumor hypoxia which is a primary driver of malignancy through its negative impact on the efficacy of radiation, immune surveillance, apoptosis, genomic stability, and accelerated angiogenesis. We performed a prospective, multicenter study to test the hypothesis that abnormal tumor vasculature and hypoxia, as measured with MRI and PET, will negatively impact survival in patients with newly diagnosed glioblastoma. Prior to the start of chemoradiation, patients with glioblastoma underwent MRI scans that included dynamic contrast enhanced and dynamic susceptibility contrast perfusion sequences to quantitate tumor cerebral blood volume/flow (CBV/CBF) and vascular permeability (ktrans) as well as 18F-Fluoromisonidazole (18F-FMISO) PET to quantitate tumor hypoxia. ROC analysis and Cox regression models were used to determine the association of imaging variables with progression-free and overall survival. Fifty patients were enrolled of which 42 had evaluable imaging data. Higher pretreatment 18F-FMISO SUVpeak (P = 0.048), mean ktrans (P = 0.024), and median ktrans (P = 0.045) were significantly associated with shorter overall survival. Higher pretreatment median ktrans (P = 0.021), normalized RCBV (P = 0.0096), and nCBF (P = 0.038) were significantly associated with shorter progression-free survival. SUVpeak [AUC = 0.75; 95% confidence interval (CI), 0.59-0.91], nRCBV (AUC = 0.72; 95% CI, 0.56-0.89), and nCBF (AUC = 0.72; 95% CI, 0.56-0.89) were predictive of survival at 1 year. Increased tumor perfusion, vascular volume, vascular permeability, and hypoxia are negative prognostic markers in newly diagnosed patients with gioblastoma, and these important physiologic markers can be measured safely and reliably using MRI and 18F-FMISO PET. Clin Cancer Res; 22(20); 5079-86. ©2016 AACR.

Piezo2 protein: A novel regulator of tumor angiogenesis and hyperpermeability.

Angiogenesis is important for invasive tumor growth and metastasis. Its inhibition is a promising tactic for limiting tumor progression. Here, we showed that Piezo2 knockdown led to decreased glioma angiogenesis and reduced vascular hyperpermeability. Piezo2 was highly expressed in tumor endothelial cells, and its knockdown suppressed vascular leakage and tumor angiogenesis. In a retinal vasculature development assay, corneal angiogenesis assay and a modified Miles assay, Piezo2 knockdown obviously decreased angiogenesis and vascular hyperpermeability. In vitro assays revealed that Piezo2 knockdown inhibited endothelial cell proliferation, migration, and tube formation. Moreover, In vitro co-culture system assay showed that Piezo2 knockdown in endothelial cells suppressed cell proliferation, migration, and invasion of glioma tumor cells. Piezo2 could regulate glioma angiogenesis via Ca2+/Wnt11/β-catenin signaling in endothelial cells. Taken together, these studies provide the evidence for Piezo2 as a critical regulator of tumor angiogenesis and vascular permeability.

Extended Time-lapse Intravital Imaging of Real-time Multicellular Dynamics in the Tumor Microenvironment.

In the tumor microenvironment, host stromal cells interact with tumor cells to promote tumor progression, angiogenesis, tumor cell dissemination and metastasis. Multicellular interactions in the tumor microenvironment can lead to transient events including directional tumor cell motility and vascular permeability. Quantification of tumor vascular permeability has frequently used end-point experiments to measure extravasation of vascular dyes. However, due to the transient nature of multicellular interactions and vascular permeability, the kinetics of these dynamic events cannot be discerned. By labeling cells and vasculature with injectable dyes or fluorescent proteins, high-resolution time-lapse intravital microscopy has allowed the direct, real-time visualization of transient events in the tumor microenvironment. Here we describe a method for using multiphoton microscopy to perform extended intravital imaging in live mice to directly visualize multicellular dynamics in the tumor microenvironment. This method details cellular labeling strategies, the surgical preparation of a mammary skin flap, the administration of injectable dyes or proteins by tail vein catheter and the acquisition of time-lapse images. The time-lapse sequences obtained from this method facilitate the visualization and quantitation of the kinetics of cellular events of motility and vascular permeability in the tumor microenvironment.

Extended Time-lapse Intravital Imaging of Real-time Multicellular Dynamics in the Tumor Microenvironment

In the tumor microenvironment, host stromal cells interact with tumor cells to promote tumor progression, angiogenesis, tumor cell dissemination and metastasis. Multicellular interactions in the tumor microenvironment can lead to transient events including directional tumor cell motility and vascular permeability. Quantification of tumor vascular permeability has frequently used end-point experiments to measure extravasation of vascular dyes. However, due to the transient nature of multicellular interactions and vascular permeability, the kinetics of these dynamic events cannot be discerned. By labeling cells and vasculature with injectable dyes or fluorescent proteins, high-resolution time-lapse intravital microscopy has allowed the direct, real-time visualization of transient events in the tumor microenvironment. Here we describe a method for using multiphoton microscopy to perform extended intravital imaging in live mice to directly visualize multicellular dynamics in the tumor microenvironment. This method details cellular labeling strategies, the surgical preparation of a mammary skin flap, the administration of injectable dyes or proteins by tail vein catheter and the acquisition of time-lapse images. The time-lapse sequences obtained from this method facilitate the visualization and quantitation of the kinetics of cellular events of motility and vascular permeability in the tumor microenvironment.

Implications of Increase in Vascular Permeability in Tumors by VEGF: A Commentary on the Pioneering Work of Harold Dvorak.

See related article by Senger et al., Cancer Res 1986;46:5629-32Visit the Cancer Research 75(th) Anniversary timeline.

A pilot study using dynamic contrast enhanced-MRI as a response biomarker of the radioprotective effect of memantine in patients receiving whole brain radiotherapy.

PurposeThis pilot prospective study sought to determine whether dynamic contrast enhanced MRI (DCE-MRI) could be used as a clinical imaging biomarker of tissue toxicity from whole brain radiotherapy (WBRT).Method14 patients who received WBRT were imaged using dynamic contrast enhanced DCE-MRI prior to and at 8-weeks, 16-weeks and 24-weeks after the initiation of WBRT. Twelve of the patients were also enrolled in the RTOG 0614 trial, which randomized patients to the use of placebo or memantine. After the unblinding of the treatments received by RTOG 0614 patients, DCE-MRI measures of tumor tissue and normal appearing white matter (NAWM) vascular permeability (Initial Area Under the Curve (AUC) Blood Adjusted) was analyzed. Cognitive, quality-of-life (QOL) assessment and blood samples were collected according to the patient's ability to tolerate the exams. Circulating endothelial cells (CEC) were measured using flow cytometry.ResultsFollowing WBRT, there was an increasing trend in the vascular permeability of tumors (p=0.09) and NAWM (p=0.06) with time. Memantine significantly (p=0.01) reduced NAWM AUC changes following radiotherapy. Patients on memantine retained (COWA p= 0.03) better cognitive functions than those on placebo. No association was observed between the level of CEC and DCE-MRI changes, time from radiotherapy or memantine use.ConclusionsDCE-MRI can detect vascular damage secondary to WBRT. Our data suggests that memantine reduces WBRT-induced brain vasculature damages.

Tumor Vascular Permeability Pattern Is Associated With Complete Response in Immunocompetent Patients With Newly Diagnosed Primary Central Nervous System Lymphoma: Retrospective Cohort Study.

A dynamic contrast-enhanced MR imaging (DCE-MRI) could provide the information about tumor drug delivery efficacy. We investigated the potential utility of the permeability pattern of DCE-MRI for predicting tumor response to high dose-methotrexate treatment and progression-free survival (PFS) in patients with primary CNS lymphoma (PCNSL). Clinical and conventional imaging parameters were assessed as potential predictors of tumor response in 48 immunocompetent PCNSL patients in a preliminary study. Fifty additional immunocompetent patients (27 men and 23 women; mean age, 60.6 years) with PCNSL underwent DCE-MRI before starting first-line treatment with high dose-methotrexate. The DCE-MRI pattern was categorized as diffuse or nondiffuse. After 4 courses of high dose methotrexate, patients underwent follow-up brain MR imaging to identify their complete response (CR). Predictors of CR and PFS were analyzed using clinical parameters, conventional MRI, and DCE-MRI. CR was noted in 20 (74.1%) of 27 patients with diffuse DCE-MRI pattern and in 4 (17.4%) of 23 patients with nondiffuse DCE-MRI pattern. The diffuse DCE-MRI pattern showed a significantly higher association with CR than the nondiffuse pattern (P < 0.001). Multivariate Cox proportional hazards model revealed that the DCE-MRI pattern (hazard ratio = 0.70; P = 0.045), age (hazard ratio = 1.47; P = 0.041), and adjuvant autologous stem-cell transplantation (hazard ratio = 6.97; P = 0.003) tended to be associated with a PFS. The pretreatment diffuse DCE-MRI pattern can be used as a potential imaging biomarker for predicting CR and a longer PFS in patients with newly diagnosed PCNSLs.

Combining the single-walled carbon nanotubes with low voltage electrical stimulation to improve accumulation of nanomedicines in tumor for effective cancer therapy

Effective delivery of biomolecules or functional nanoparticles into target sites has always been the primary objective for cancer therapy. We demonstrated that by combining single-walled carbon nanotubes (SWNTs) with low-voltage (LV) electrical stimulation, biomolecule delivery can be effectively enhanced through reversible electroporation (EP). Clear pore formation in the cell membrane is observed due to LV (50V) pulse electrical stimulation amplified by SWNTs. The cell morphology remains intact and high cell viability is retained. This modality of SWNT + LV pulses can effectively transfer both small molecules and macromolecules into cells through reversible EP. The results of animal studies also suggest that treatment with LV pulses alone cannot increase vascular permeability in tumors unless after the injection of SWNTs. The nanoparticles can cross the permeable vasculature, which enhances their accumulation in the tumor tissue. Therefore, in cancer treatment, both SWNT + LV pulse treatment followed by the injection of LIPO-DOX® and SWNT/DOX + LV pulse treatment can increase tumor inhibition and delay tumor growth. This novel treatment modality applied in a human cancer xenograft model can provide a safe and effective therapy using various nanomedicines in cancer treatment.

Tumor-Priming Smoothened Inhibitor Enhances Deposition and Efficacy of Cytotoxic Nanoparticles in a Pancreatic Cancer Model.

Most pancreatic adenocarcinoma patients present with unresectable disease and benefit little from chemotherapy. Poor tumor perfusion and vascular permeability limit drug deposition. Previous work showed that Smoothened inhibitors of hedgehog signaling (sHHI) promote neovascularization in spontaneous mouse models of pancreatic cancer (PaCA) and enhance tumor permeability to low-molecular weight compounds. Here, we tested the hypothesis that sHHI can enhance tumor deposition and efficacy of drug-containing nanoparticles consisting of 80 to 100 nm sterically-stabilized liposomes (SSL) containing doxorubicin (SSL-DXR). SCID mice bearing low-passage patient-derived PaCA xenografts (PDX) were pretreated p.o. for 10 days with 40 mg/kg/d NVP-LDE225 (erismodegib), followed by i.v. SSL-DXR. Microvessel density, permeability, perfusion, and morphology were compared with untreated controls, as was SSL deposition and therapeutic efficacy. The sHHI alone affected tumor growth minimally, but markedly increased extravasation of nanoparticles into adenocarcinoma cell-enriched regions of the tumor. Immunostaining showed that sHHI treatment decreased pericyte coverage (α-SMA(+)) of CD31(+) vascular endothelium structures, and increased the abundance of endothelium-poor (CD31(-)) basement membrane structures (collagen IV(+)), suggesting increased immature microvessels. SSL-DXR (15 mg/kg) administered after sHHI pretreatment arrested tumor volume progression and decreased tumor perfusion/permeability, suggesting an initial vascular pruning response. Compared with controls, one cycle of 10-day sHHI pretreatment followed by 6 mg/kg SSL-DXR doubled median tumor progression time. Three cycles of treatment with sHHI and SSL-DXR, with a 10-day between-cycle drug holiday, nearly tripled median tumor progression time. Based upon these data, short-term sHHI treatment sequenced with nanoparticulate drug carriers constitutes a potential strategy to enhance efficacy of pancreatic cancer therapy.

Multi-targeting Peptide-Functionalized Nanoparticles Recognized Vasculogenic Mimicry, Tumor Neovasculature, and Glioma Cells for Enhanced Anti-glioma Therapy.

Chemotherapy failure of glioma, the most aggressive and devastating cancer, might be ascribed to the physiologic barriers of the tumor mainly including heterogeneous tumor perfusion and vascular permeability, which result in a limited penetration of chemotherapeutics. Besides, the vasculogenic mimicry (VM) channels, which are highly resistant to anti-angiogenic therapy and serve as a complement of angiogenesis, were abound in glioma and always associated with tumor recurrence. In order to enhance the therapy effect of anti-glioma, we developed a PEG-PLA-based nanodrug delivery system (nanoparticles, NP) in this study and modified its surface with CK peptide, which was composed of a human sonic hedgehog (SHH) targeting peptide (CVNHPAFAC) and a KDR targeting peptide (K237) through a GYG linker, for facilitating efficient VM channels, tumor neovasculature, and glioma cells multi-targeting delivery of paclitaxel. In vitro cellular assay showed that CK-NP-PTX not only exhibited the strongest antiproliferation effect on U87MG cells and HUVEC cells but also resulted in the most efficient destruction of VM channels when compared with CVNHPAFAC-NP, K237-NP, and the unmodified ones. Besides, CK-NP accumulated more selectively at the glioma site as demonstrated by in vivo and ex vivo imaging. As expected, the glioma-bearing mice treated with CK-NP-PTX achieved the longest median survival time compared to those treated with CVNHPAFAC-NP-PTX and K237-NP-PTX. These findings indicated that the multi-targeting therapy mediated by CK peptide might provide a promising way for glioblastoma therapy.

Dynamic Measurement of Tumor Vascular Permeability and Perfusion using a Hybrid System for Simultaneous Magnetic Resonance and Fluorescence Imaging.

Assessing tumor vascular features including permeability and perfusion is essential for diagnostic and therapeutic purposes. The aim of this study was to compare fluorescence and magnetic resonance imaging (MRI)-based vascular readouts in subcutaneously implanted tumors in mice by simultaneous dynamic measurement of tracer uptake using a hybrid fluorescence molecular tomography (FMT)/MRI system. Vascular permeability was measured using a mixture of extravascular imaging agents, GdDOTA and the dye Cy5.5, and perfusion using a mixture of intravascular agents, Endorem and a fluorescent probe (Angiosense). Dynamic fluorescence reflectance imaging (dFRI) was integrated into the hybrid system for high temporal resolution. Excellent correspondence between uptake curves of Cy5.5/GdDOTA and Endorem/Angiosense has been found with correlation coefficients R > 0.98. The two modalities revealed good agreement regarding permeability coefficients and centers-of-gravity of the imaging agent distribution. The FMT/dFRI protocol presented is able to accurately map physiological processes and poses an attractive alternative to MRI for characterizing tumor neoangiogenesis.

Sonochemical internalization using 5-aminolevulinic acid enhances the cytotoxic effect of bleomycin

s / Photodiagnosis and Photodynamic Therapy 12 (2015) 325–375 343 Results: The few treated cases for the moment show fading lesions in an average of 4 sessions, including an immunocompromised patient had a very quick recovery. Conclusions:Fora country likeEcuador is very important search new cheaper and efficient alternatives for the treatment of health public problems. The results do not present major problems to raise a medical guideline, however this study will be continued and should be replicated in different demographic areas throughout the country collecting statistical data over types of patients, sessions conducted and recurrences. http://dx.doi.org/10.1016/j.pdpdt.2015.07.073 Examining the effect of liposomal lapatinib in combination with PDT for the treatment of malignant gliomas Carl Fisher1,2,3,4, Carolyn Niu1,2,3,4, Warren Foltz1,2,3,4, Girgis Obaid1,2,3,4, Tayyaba Hasan1,2,3,4, Lothar Lilge1,2,3,4 1 Department of Medical Biophysics, University of Toronto, Canada 2 Department of Radiation Oncology, University Health Network, Canada 3 Wellman Center for Photomedicine, Massachusetts General Hospital, USA 4 Division of Molecular Imaging, Princess Margaret Cancer Centre, Canada Malignant gliomas are invasive and difficult to treat tumours. Of this group of tumours, Glioblastoma Multiforme (GBM) is the most common and the most aggressive. Of proposed GBM treatments, ALA-PpIX mediated Photodynamic Therapy (PDT) has been explored and researched into. The goal of our group is to examine whether combining PDT with EGFR kinase inhibitors leads to greater outcomes compared to PDT or EGFR inhibitors alone. The study proposed utilized the human cell line U87 (EGFR positive) in combination with a liposomal based EGFR inhibitor (lapatinib) that was combined with PDT treatment. Pre-PDT animals were scannedwithaBruker7T small animalMRI systemtomeasure cerebral blood flow (CBF) and tumor vascular permeability and began treatment with the liposomal lapatinib conjugation. Post-PDT animal were scanned at regular intervals to measure CBF and tumour size between the two groups. In the initial pilot study, liposomal lapatinib combined with PDT lead to significant increases in survival compared to PDT alone and demonstrated differences in both CBF and tumour response to PDT. Ongoing studies are focused on increasing the number of animals and get quantitativemeasures of CBF and cerebral blood volume both preand post-PDT. http://dx.doi.org/10.1016/j.pdpdt.2015.07.074 Sonochemical internalization using 5-aminolevulinic acid enhances the cytotoxic effect of bleomycin Tomohiro Osaki1, Yoshihiro Uto2, Masahiro Ishizuka3, Toru Tanaka3, Nobuyasu Yamanaka4, Tsukasa Kurahashi4, Kazuo Azuma1, Yusuke Murahata1, Takeshi Tsuka1, Norihiko Ito1, Tomohiro Imagawa1, Yoshiharu Okamoto1 1 Department of Veterinary Clinical Medicine, School of Veterinary Medicine, Tottori University, Japan 2 Department of Life System, Institute of Technology and Science, Graduate School, Tokushima University, Japan 3 SBI Pharmaceuticals Co., Ltd., Japan 4 ITO Physiotherapy & Rehabilitation, Japan Photochemical internalization (PCI) is a technology to improve the intracellular delivery ofmacromolecules andbasedon the same principle as photodynamic therapy (PDT). PCI is based on the use of photosensitizers located in endocytic vesicles that upon activation by light induce rupture of endocytic vesicles and release the macromolecules into cytosol. Bleomycin (BLM) is used for chemotherapeutic agent for PCI. The limitation of PDT and PCI is the penetration of light in the tissue. It is difficult to treat deepseated tumor with PDT and PCI. Sonodynamic therapy (SDT) is also an antitumor strategy that kills tumor cells by ultrasound and photosensitizer. Ultrasound can penetrate deep tissue and excite photosensitizer. We hypothesized that ultrasound could excite photosensitizers used for PCI, and this newmethod could improve the intracellular delivery of macromolecules. We defined this new method as sonochemical internalization (SCI). In this study, we examined whether internalization of BLM by ultrasound and 5aminolevulinic acid (5-ALA) was able to destroy tumors. In vitro and in vivo study, a significant cytotoxic effect of SCI of BLM using 5-ALA was observed. It was considered that SCI might act as an ultrasound-directed drug-delivery system by triggered release of endocytosed macromolecules into cytosol as PCI. http://dx.doi.org/10.1016/j.pdpdt.2015.07.075 Acridine orange as radiosensitizer to improve cell damage caused by radiotherapy in breast cancer cells line Cintia Teles de Andrade, Jose Dirceu Vollet-Filho, Layla Pires, Vanderlei Salvador Bagnato, Cristina Kurachi Sao Carlos Institute of Physics, University of Sao

Examining the effect of liposomal lapatinib in combination with PDT for the treatment of malignant gliomas

s / Photodiagnosis and Photodynamic Therapy 12 (2015) 325–375 343 Results: The few treated cases for the moment show fading lesions in an average of 4 sessions, including an immunocompromised patient had a very quick recovery. Conclusions:Fora country likeEcuador is very important search new cheaper and efficient alternatives for the treatment of health public problems. The results do not present major problems to raise a medical guideline, however this study will be continued and should be replicated in different demographic areas throughout the country collecting statistical data over types of patients, sessions conducted and recurrences. http://dx.doi.org/10.1016/j.pdpdt.2015.07.073 Examining the effect of liposomal lapatinib in combination with PDT for the treatment of malignant gliomas Carl Fisher1,2,3,4, Carolyn Niu1,2,3,4, Warren Foltz1,2,3,4, Girgis Obaid1,2,3,4, Tayyaba Hasan1,2,3,4, Lothar Lilge1,2,3,4 1 Department of Medical Biophysics, University of Toronto, Canada 2 Department of Radiation Oncology, University Health Network, Canada 3 Wellman Center for Photomedicine, Massachusetts General Hospital, USA 4 Division of Molecular Imaging, Princess Margaret Cancer Centre, Canada Malignant gliomas are invasive and difficult to treat tumours. Of this group of tumours, Glioblastoma Multiforme (GBM) is the most common and the most aggressive. Of proposed GBM treatments, ALA-PpIX mediated Photodynamic Therapy (PDT) has been explored and researched into. The goal of our group is to examine whether combining PDT with EGFR kinase inhibitors leads to greater outcomes compared to PDT or EGFR inhibitors alone. The study proposed utilized the human cell line U87 (EGFR positive) in combination with a liposomal based EGFR inhibitor (lapatinib) that was combined with PDT treatment. Pre-PDT animals were scannedwithaBruker7T small animalMRI systemtomeasure cerebral blood flow (CBF) and tumor vascular permeability and began treatment with the liposomal lapatinib conjugation. Post-PDT animal were scanned at regular intervals to measure CBF and tumour size between the two groups. In the initial pilot study, liposomal lapatinib combined with PDT lead to significant increases in survival compared to PDT alone and demonstrated differences in both CBF and tumour response to PDT. Ongoing studies are focused on increasing the number of animals and get quantitativemeasures of CBF and cerebral blood volume both preand post-PDT. http://dx.doi.org/10.1016/j.pdpdt.2015.07.074 Sonochemical internalization using 5-aminolevulinic acid enhances the cytotoxic effect of bleomycin Tomohiro Osaki1, Yoshihiro Uto2, Masahiro Ishizuka3, Toru Tanaka3, Nobuyasu Yamanaka4, Tsukasa Kurahashi4, Kazuo Azuma1, Yusuke Murahata1, Takeshi Tsuka1, Norihiko Ito1, Tomohiro Imagawa1, Yoshiharu Okamoto1 1 Department of Veterinary Clinical Medicine, School of Veterinary Medicine, Tottori University, Japan 2 Department of Life System, Institute of Technology and Science, Graduate School, Tokushima University, Japan 3 SBI Pharmaceuticals Co., Ltd., Japan 4 ITO Physiotherapy & Rehabilitation, Japan Photochemical internalization (PCI) is a technology to improve the intracellular delivery ofmacromolecules andbasedon the same principle as photodynamic therapy (PDT). PCI is based on the use of photosensitizers located in endocytic vesicles that upon activation by light induce rupture of endocytic vesicles and release the macromolecules into cytosol. Bleomycin (BLM) is used for chemotherapeutic agent for PCI. The limitation of PDT and PCI is the penetration of light in the tissue. It is difficult to treat deepseated tumor with PDT and PCI. Sonodynamic therapy (SDT) is also an antitumor strategy that kills tumor cells by ultrasound and photosensitizer. Ultrasound can penetrate deep tissue and excite photosensitizer. We hypothesized that ultrasound could excite photosensitizers used for PCI, and this newmethod could improve the intracellular delivery of macromolecules. We defined this new method as sonochemical internalization (SCI). In this study, we examined whether internalization of BLM by ultrasound and 5aminolevulinic acid (5-ALA) was able to destroy tumors. In vitro and in vivo study, a significant cytotoxic effect of SCI of BLM using 5-ALA was observed. It was considered that SCI might act as an ultrasound-directed drug-delivery system by triggered release of endocytosed macromolecules into cytosol as PCI. http://dx.doi.org/10.1016/j.pdpdt.2015.07.075 Acridine orange as radiosensitizer to improve cell damage caused by radiotherapy in breast cancer cells line Cintia Teles de Andrade, Jose Dirceu Vollet-Filho, Layla Pires, Vanderlei Salvador Bagnato, Cristina Kurachi Sao Carlos Institute of Physics, University of Sao

Treatment of condyloma acuminatum using methyl-aminolaevulinic acid for introduce a medical guideline on health public system

s / Photodiagnosis and Photodynamic Therapy 12 (2015) 325–375 343 Results: The few treated cases for the moment show fading lesions in an average of 4 sessions, including an immunocompromised patient had a very quick recovery. Conclusions:Fora country likeEcuador is very important search new cheaper and efficient alternatives for the treatment of health public problems. The results do not present major problems to raise a medical guideline, however this study will be continued and should be replicated in different demographic areas throughout the country collecting statistical data over types of patients, sessions conducted and recurrences. http://dx.doi.org/10.1016/j.pdpdt.2015.07.073 Examining the effect of liposomal lapatinib in combination with PDT for the treatment of malignant gliomas Carl Fisher1,2,3,4, Carolyn Niu1,2,3,4, Warren Foltz1,2,3,4, Girgis Obaid1,2,3,4, Tayyaba Hasan1,2,3,4, Lothar Lilge1,2,3,4 1 Department of Medical Biophysics, University of Toronto, Canada 2 Department of Radiation Oncology, University Health Network, Canada 3 Wellman Center for Photomedicine, Massachusetts General Hospital, USA 4 Division of Molecular Imaging, Princess Margaret Cancer Centre, Canada Malignant gliomas are invasive and difficult to treat tumours. Of this group of tumours, Glioblastoma Multiforme (GBM) is the most common and the most aggressive. Of proposed GBM treatments, ALA-PpIX mediated Photodynamic Therapy (PDT) has been explored and researched into. The goal of our group is to examine whether combining PDT with EGFR kinase inhibitors leads to greater outcomes compared to PDT or EGFR inhibitors alone. The study proposed utilized the human cell line U87 (EGFR positive) in combination with a liposomal based EGFR inhibitor (lapatinib) that was combined with PDT treatment. Pre-PDT animals were scannedwithaBruker7T small animalMRI systemtomeasure cerebral blood flow (CBF) and tumor vascular permeability and began treatment with the liposomal lapatinib conjugation. Post-PDT animal were scanned at regular intervals to measure CBF and tumour size between the two groups. In the initial pilot study, liposomal lapatinib combined with PDT lead to significant increases in survival compared to PDT alone and demonstrated differences in both CBF and tumour response to PDT. Ongoing studies are focused on increasing the number of animals and get quantitativemeasures of CBF and cerebral blood volume both preand post-PDT. http://dx.doi.org/10.1016/j.pdpdt.2015.07.074 Sonochemical internalization using 5-aminolevulinic acid enhances the cytotoxic effect of bleomycin Tomohiro Osaki1, Yoshihiro Uto2, Masahiro Ishizuka3, Toru Tanaka3, Nobuyasu Yamanaka4, Tsukasa Kurahashi4, Kazuo Azuma1, Yusuke Murahata1, Takeshi Tsuka1, Norihiko Ito1, Tomohiro Imagawa1, Yoshiharu Okamoto1 1 Department of Veterinary Clinical Medicine, School of Veterinary Medicine, Tottori University, Japan 2 Department of Life System, Institute of Technology and Science, Graduate School, Tokushima University, Japan 3 SBI Pharmaceuticals Co., Ltd., Japan 4 ITO Physiotherapy & Rehabilitation, Japan Photochemical internalization (PCI) is a technology to improve the intracellular delivery ofmacromolecules andbasedon the same principle as photodynamic therapy (PDT). PCI is based on the use of photosensitizers located in endocytic vesicles that upon activation by light induce rupture of endocytic vesicles and release the macromolecules into cytosol. Bleomycin (BLM) is used for chemotherapeutic agent for PCI. The limitation of PDT and PCI is the penetration of light in the tissue. It is difficult to treat deepseated tumor with PDT and PCI. Sonodynamic therapy (SDT) is also an antitumor strategy that kills tumor cells by ultrasound and photosensitizer. Ultrasound can penetrate deep tissue and excite photosensitizer. We hypothesized that ultrasound could excite photosensitizers used for PCI, and this newmethod could improve the intracellular delivery of macromolecules. We defined this new method as sonochemical internalization (SCI). In this study, we examined whether internalization of BLM by ultrasound and 5aminolevulinic acid (5-ALA) was able to destroy tumors. In vitro and in vivo study, a significant cytotoxic effect of SCI of BLM using 5-ALA was observed. It was considered that SCI might act as an ultrasound-directed drug-delivery system by triggered release of endocytosed macromolecules into cytosol as PCI. http://dx.doi.org/10.1016/j.pdpdt.2015.07.075 Acridine orange as radiosensitizer to improve cell damage caused by radiotherapy in breast cancer cells line Cintia Teles de Andrade, Jose Dirceu Vollet-Filho, Layla Pires, Vanderlei Salvador Bagnato, Cristina Kurachi Sao Carlos Institute of Physics, University of Sao