Molecular Bioluminescence Imaging as a Noninvasive Tool for Monitoring Tumor Growth and Therapeutic Response to MRI-Guided Laser Ablation in a Rat Model of Hepatocellular Carcinoma

Thompson, Scott M.; Callstrom, Matthew R.; Knudsen, Bruce E.; Anderson, J. L.; Sutor, Shari L.; Butters, Kim A.; Kuo, Chaincy; Grande, Joseph P.; Roberts, Lewis R.; Woodrum, David A.

doi:10.1097/rli.0b013e31827a4a3f

Cited by 22 publications

(24 citation statements)

References 44 publications

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“…N1S1 cells were stably transfected with a pGL4.51[ luc2 /CMV/Neo] luciferase reporter vector (Promega) using X‐tremeGENE HP DNA Transfection reagent (Roche) followed by positive selection in G418 (Life Technologies, Carlsbad, CA) and establishment of clonal populations using limiting dilution method as previously described .…”

Section: Methodsmentioning

confidence: 99%

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Heat stress induced cell death mechanisms in hepatocytes and hepatocellular carcinoma: In vitro and in vivo study

et al. 2014

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Background and Objective The aims of the present study were to investigate the thermal-dose dependent effect of heat stress on hepatocyte and HCC cell death mechanisms using clinically relevant experimental heat stress conditions in vitro and to investigate apoptotic cell death induced by laser thermal ablation in vivo. Study Design/Materials and Methods Institutional Animal Care and Use Committee approved all studies. Hepatocyte and HCC cell lines were heat stressed from 37°C to 60°C for 2 or 10 minutes and assessed for viability, cytotoxicity and caspase-3/7 activity at 6 and/or 24 hours post-treatment (N=3). Viability experiments were repeated with the RIPK1 inhibitor Necrostatin-1 to block necroptosis (N=3). Rats with orthotopic HCC tumors stably expressing luciferase (N1S1luc2) were randomized to US-guided laser ablation (3W-45s for an intentional partial ablation; N=6) or sham (N=6) and followed by post-ablation caspase-3/7 bioluminescence imaging at 6 and 24 hours and cleaved caspase-3 immunostaining. p<0.05 was considered statistically significant. Results Heat-stress induced apoptosis and necrosis in hepatocytes and HCC cells in a thermal dose and cell-type dependent manner. Inhibition of RIPIK1-mediated necroptosis induced a significant, differential increase in HCC cell viability under physiologic and hyperthermic heat stress (p<0.001). Intentional partial laser thermal ablation induced a significant increase in caspase-3/7 activity in the laser versus sham ablation groups at both 6 hours (10.1-fold, p<0.01) and 24 hours (16.7 fold, p<0.02). Immunohistochemistry confirmed increased cleaved caspase-3 staining at the tumor ablation margin 24 hours post-ablation. Conclusions Both regulated and non-regulated cell death mechanisms mediate heat stress-induced HCC cell killing and vary between hepatocytes and HCC subtypes. Apoptosis is a significant mechanism of cell death at the HCC tumor ablation margin.

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Section: Methodsmentioning

confidence: 99%

“…Studies were approved by the Institutional Animal Care and Use Committee. N1S1 orthotopic HCC model was developed as previously described using N1S1 luc2 cells stably expressing firefly luciferase ( N = 12) .…”

Section: Methodsmentioning

confidence: 99%

Heat stress induced cell death mechanisms in hepatocytes and hepatocellular carcinoma: In vitro and in vivo study

et al. 2014

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“…Transfection protocol.-HCC lines stably expressing firefly luciferase were generated by using methods previously described (Appendix E1 [online]) (25). A standard curve was generated for both the N1S1luc2 and AS30Dluc2 cell lines to estimate cell number in vitro ( Fig E1 [online]).…”

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“…This model was adapted from Huang et al (26). Proliferation of the non-heat-stressed HCC cells (N1S1luc2, AS30Dluc2) was monitored with bioluminescence imaging from 0 to 6 days postplating (25). To assess the role of PI3K/mTOR signaling in this model, non-heatstressed cells were pretreated for 1 hour with the small molecule inhibitor NVP-BEZ25 (1 mM) before culture with their respective feeder layers.…”

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confidence: 99%

“…The N1S1 orthotopic HCC model was developed in male Sprague-Dawley rats (Harlan/Envigo Laboratories, Indianapolis, Indiana) (weight range, 400-500 g) (27). Immediately after injection of N1S1 cells (3 3 10 6 cells) into the median hepatic lobe, rats were randomly assigned to undergo hepatic laser ablation in the HCC cell-bearing median hepatic lobe (local) or left lateral hepatic lobe (distant) with an energy dose of 3 W for 60 or 90 seconds (hereafter, 3W360s and 3W390s, respectively) or sham ablation with a Food and Drug Administration-approved 980-nm laser generator (Visualase, Houston, Tex) (six or more rats per experimental group) (24,25,27). The laser fiber was inserted but not activated in sham-ablated animals.…”

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Heat Stress and Hepatic Laser Thermal Ablation Induce Hepatocellular Carcinoma Growth: Role of PI3K/mTOR/AKT Signaling

et al. 2018

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Purpose To determine if heat stress and hepatic laser thermal ablation induce hepatocellular carcinoma (HCC) growth and to identify growth factors induced by heat stress. Materials and Methods Non-heat-stressed HCC cells were cocultured with HCC cells or hepatocytes that were heat stressed at 37°C (physiologic), 45°C (moderate), or 50°C (severe) for 10 minutes and proliferation monitored with bioluminescence imaging for up to 6 days after heat stress (three experiments). Rats bearing orthotopic N1S1 HCC were randomly assigned to undergo immediate sham or laser thermal (3 W for 60 or 90 seconds; hereafter, 3W×60s and 3W×90s, respectively) ablation of the median (local) or left (distant) hepatic lobe, and tumor growth was monitored with magnetic resonance imaging for up to 18 days after ablation (six or more rats per group). Experiments were repeated with rats randomly assigned to receive either the adjuvant phosphoinositide 3-kinase (PI3K)/mammalian target of rapamycin (mTOR) inhibitor (NVP-BEZ235) or the vehicle control. Heat-stressed HCC cells and hepatocytes were analyzed by using microarray or quantitative real-time polymerase chain reaction analysis for growth factor expression (three or more experiments). Groups were compared by using one- or two-way analysis of variance, and post hoc pairwise comparison was performed with the Dunnett test. Results There were more non-heat-stressed HCC cells when cells were cocultured with cells subjected to moderate but not physiologic or severe heat stress (P < .001 for both). Local intrahepatic N1S1 tumors were larger at day 18 in the 3W×60s (mean, 3102 mm ± 463 [standard error]; P = .004) and 3W×90s (mean, 3538 mm ± 667; P < .001) groups than in the sham group (mean, 1363 mm ± 361) but not in distant intrahepatic tumors (P = .31). Adjuvant BEZ235 resulted in smaller N1S1 tumors in the BEZ235 and laser thermal ablation group than in the vehicle control and laser thermal ablation group (mean, 1731 mm ± 1457 vs 3844 mm ± 2400, P < .001). Moderate heat stress induced expression of growth factors in HCC cells and hepatocytes, including heparin-binding growth factor, fibroblast growth factor 21, and nerve growth factor (range, 2.9-66.9-fold; P < .05). Conclusion Moderate heat stress and laser thermal ablation induce hepatocellular carcinoma growth, which is prevented with adjuvant PI3K/mTOR/protein kinase B inhibition.

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Firefly Luciferin‐Inspired Biocompatible Chemistry for Protein Labeling and In Vivo Imaging

Wang

Luo

et al. 2017

Chemistry A European J

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Biocompatible reactions have emerged as versatile tools to build various molecular imaging probes that hold great promise for the detection of biological processes in vitro and/or in vivo. In this Minireview, we describe the recent advances in the development of a firefly luciferin-inspired biocompatible reaction between cyanobenzothiazole (CBT) and cysteine (Cys), and highlight its versatility to label proteins and build multimodality molecular imaging probes. The review starts from the general introduction of biocompatible reactions, which is followed by briefly describing the development of the firefly luciferin-inspired biocompatible chemistry. We then discuss its applications for the specific protein labeling and for the development of multimodality imaging probes (fluorescence, bioluminescence, MRI, PET, photoacoustic, etc.) that enable high sensitivity and spatial resolution imaging of redox environment, furin and caspase-3/7 activity in living cells and mice. Finally, we offer the conclusions and our perspective on the various and potential applications of this reaction. We hope that this review will contribute to the research of biocompatible reactions for their versatile applications in protein labeling and molecular imaging.

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Molecular Bioluminescence Imaging as a Noninvasive Tool for Monitoring Tumor Growth and Therapeutic Response to MRI-Guided Laser Ablation in a Rat Model of Hepatocellular Carcinoma

Cited by 22 publications

References 44 publications

Heat stress induced cell death mechanisms in hepatocytes and hepatocellular carcinoma: In vitro and in vivo study

Heat stress induced cell death mechanisms in hepatocytes and hepatocellular carcinoma: In vitro and in vivo study

Heat Stress and Hepatic Laser Thermal Ablation Induce Hepatocellular Carcinoma Growth: Role of PI3K/mTOR/AKT Signaling

Firefly Luciferin‐Inspired Biocompatible Chemistry for Protein Labeling and In Vivo Imaging

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