A Fas Ligand (FasL)-Fused Humanized Antibody Against Tumor-Associated Glycoprotein 72 Selectively Exhibits the Cytotoxic Effect Against Oral Cancer Cells with a Low FasL/Fas Ratio

Chien, Ming Hsien; Chang, Wei‐Min; Lee, Wei Jiunn; Chang, Yu Chan; Lai, Tsung‐Ching; Chan, Derek V.; Sharma, Rahul; Lin, Yuan; Hsiao, Michael

doi:10.1158/1535-7163.mct-16-0314

Cited by 18 publications

(19 citation statements)

References 46 publications

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“…NA cell line was established as a fibronectin-producing cell line (14). Furthermore, it has been reported that the cytotoxicity of anti-cancer reagents in OSCC has been evaluated in SQUU-A cell line and SQUU-B cell line (15), SAS cell line (16), NA cell line (17) using various conventional methods. That is why, we selected four these cell lines with each characteristic feature in the present study, which were also used in cytotoxic assay in previous study.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of IC50 levels immediately after treatment with anticancer reagents using a real‑time cell monitoring device

et al. 2019

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A real-time cell-monitoring analysis (RTCA) system was previously developed based on the change in impedance when cells attach and spread in a culture dish coated with a gold microelectrode array. However, the potential applications of this system have not yet been fully demonstrated. The purpose of this study was to test the utility of the RTCA system to determine the cytotoxicity of four anticancer agents in carcinoma cells. The results were compared with those of the conventional WST-8 assay at the endpoint to determine the potential of the RTCA system as a new real-time assay method to evaluate cytotoxicity. iCELLigence was used as the RTCA system in this study. Suspensions of oral squamous cell carcinoma (OSCC) cell lines were seeded (2x10 4 cells/well) onto the E-plate (the culture plate of the iCELLigence system). After 24 h of culture, anticancer agents were added to each well, and changes in electrical impedance (cell index, CI) were recorded for another 72 h of culture. Cell proliferation was detected in real-time by the RTCA device in an automated, high throughput manner. Then, the IC 50 profiles of the four anticancer agents were calculated based on the real-time cell index values. The results indicated that the RTCA system was useful in evaluating cytotoxic reactions immediately after the addition of the anticancer agents as it was able to record the data in real-time. Furthermore, the IC 50 levels measured by the real-time assay were lower than those measured by the endpoint assay. Thus, RTCA systems can be used to evaluate chemotherapeutic agents in cancer cells as well as their side effects in normal cells.

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Section: Introductionmentioning

confidence: 99%

Evaluation of IC50 levels immediately after treatment with anticancer reagents using a real‑time cell monitoring device

et al. 2019

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“…However, the specific activity of the hFasLECD sample was at least 20 times higher than an anti-mouse FasR agonistic monoclonal antibody, Jo2, in inducing apoptosis against FasR overexpressing mouse cells, and showed much less toxicity with regard to the liver failure in vivo [ 5 ]. To overcome the above mentioned problem, numerous studies for delivering the protein specifically toward the target cells have been made by exploiting the gene-fusion technology using the genes of single chain variable fragments of the cell-surface antigen recognizing antibodies and the extracellular domains of cytokines as the fusion components [ 6 – 10 ]. On the other hand, the administration of many cytotoxic drugs, including the ones in clinical uses, is known to significantly affect the number of cell-surface hFasR, which determines the susceptibility to apoptosis execution by hFasL [ 11 – 14 ].…”

Section: Introductionmentioning

confidence: 99%

Site-specific chemical conjugation of human Fas ligand extracellular domain using trans-cyclooctene – methyltetrazine reactions

Muraki

Hirota

2017

BMC Biotechnol

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BackgroundFas ligand plays a key role in the human immune system as a major cell death inducing protein. The extracellular domain of human Fas ligand (hFasLECD) triggers apoptosis of malignant cells, and therefore is expected to have substantial potentials in medical biotechnology. However, the current application of this protein to clinical medicine is hampered by a shortage of the benefits relative to the drawbacks including the side-effects in systemic administration. Effective procedures for the engineering of the protein by attaching useful additional functions are required to overcome the problem.ResultsA procedure for the site-specific chemical conjugation of hFasLECD with a fluorochrome and functional proteins was devised using an inverse-electron-demand Diels-Alder reaction between trans-cyclooctene group and methyltetrazine group. The conjugations in the present study were attained by using much less molar excess amounts of the compounds to be attached as compared with the conventional chemical modification reactions using maleimide derivatives in the previous study. The isolated conjugates of hFasLECD with sulfo-Cy3, avidin and rabbit IgG Fab’ domain presented the functional and the structural integrities of the attached molecules without impairing the specific binding activity toward human Fas receptor extracellular domain.ConclusionsThe present study provided a new fundamental strategy for the production of the engineered hFasLECDs with additional beneficial functions, which will lead to the developments of the improved diagnostic systems and the effective treatment methods of serious diseases by using this protein as a component of novel molecular tools.Electronic supplementary materialThe online version of this article (doi:10.1186/s12896-017-0381-2) contains supplementary material, which is available to authorized users.

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“…There is a substantial translational potential for our understanding of the roles and regulation of various forms of released Fas ligand. This includes development of novel approaches to treatment with, for example, FasL-fused humanized antibodies to sensitize target cells to cell death [ 108 , 109 , 110 ] as suggested in glaucoma treatment [ 111 , 112 ], novel forms of RNA therapy [ 113 ], prognosis of long-term allergic outcomes at birth [ 114 ] or even for schizophrenia treatment [ 115 ]. However, there remain a number of outstanding problems that need to be addressed with regard to soluble Fas ligand as a therapeutic target.…”

Section: Discussionmentioning

confidence: 99%

sFasL—The Key to a Riddle: Immune Responses in Aging Lung and Disease

Wallach-Dayan

Petukhov

Ahdut-HaCohen

et al. 2021

IJMS

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By dint of the aging population and further deepened with the Covid-19 pandemic, lung disease has turned out to be a major cause of worldwide morbidity and mortality. The condition is exacerbated when the immune system further attacks the healthy, rather than the diseased, tissue within the lung. Governed by unremittingly proliferating mesenchymal cells and increased collagen deposition, if inflammation persists, as frequently occurs in aging lungs, the tissue develops tumors and/or turns into scars (fibrosis), with limited regenerative capacity and organ failure. Fas ligand (FasL, a ligand of the Fas cell death receptor) is a key factor in the regulation of these processes. FasL is primarily found in two forms: full length (membrane, or mFasL) and cleaved (soluble, or sFasL). We and others found that T-cells expressing the mFasL retain autoimmune surveillance that controls mesenchymal, as well as tumor cell accumulation following an inflammatory response. However, mesenchymal cells from fibrotic lungs, tumor cells, or cells from immune-privileged sites, resist FasL+ T-cell-induced cell death. The mechanisms involved are a counterattack of immune cells by FasL, by releasing a soluble form of FasL that competes with the membrane version, and inhibits their cell death, promoting cell survival. This review focuses on understanding the previously unrecognized role of FasL, and in particular its soluble form, sFasL, in the serum of aged subjects, and its association with the evolution of lung disease, paving the way to new methods of diagnosis and treatment.

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A Fas Ligand (FasL)-Fused Humanized Antibody Against Tumor-Associated Glycoprotein 72 Selectively Exhibits the Cytotoxic Effect Against Oral Cancer Cells with a Low FasL/Fas Ratio

Cited by 18 publications

References 46 publications

Evaluation of IC50 levels immediately after treatment with anticancer reagents using a real‑time cell monitoring device

Evaluation of IC50 levels immediately after treatment with anticancer reagents using a real‑time cell monitoring device

Site-specific chemical conjugation of human Fas ligand extracellular domain using trans-cyclooctene – methyltetrazine reactions

sFasL—The Key to a Riddle: Immune Responses in Aging Lung and Disease

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