Penetration and Binding of Monoclonal Antibody in Human Osteosarcoma Multicell Spheroids: Comparison of confocal laser scanning microscopy and autoradiography

Hjelstuen, M.; Rasch-Halvorsen, Kathrine; Brekken, Christian; Bruland, Øyvind S.; Davies, Catharina de Lange

doi:10.3109/02841869609101641

Cited by 16 publications

(14 citation statements)

References 20 publications

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“…By incubating spheroids with an anticancer drug, it then becomes possible to examine the kinetics of drug penetration. Unlike a previous method12, the protocol described here is relatively simple and straightforward to learn, rapid in producing samples, and economically feasible.…”

Section: Introductionmentioning

confidence: 99%

Rapid Generation of In Vitro Multicellular Spheroids for the Study of Monoclonal Antibody Therapy

Phung¹,

Barbone²,

Broaddus³

et al. 2011

J. Cancer

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Tumor microenvironments present significant barriers to penetration by antibodies and immunoconjugates and are difficult to study in vitro. Cells cultured as monolayers typically exhibit less resistance to therapy than those grown in vivo. Therefore, it is important to develop an alternative research model that better represents in vivo tumors. We have developed a protocol to produce multicellular spheroids, a simple and more relevant model of in vivo tumors that allows for further investigations of the microenvironmental effects on drug penetration and tumor cell killing. The protocol is used to produce in vitro three-dimensional tumor spheroids from established human cancer cell lines and primary cancer cells isolated from patients without the use of any extracellular components. To study the ability of tumor-targeting immunoconjugates to penetrate these tumor spheroids in vitro, we have used an immunotoxin targeting mesothelin, a surface protein expressed in malignant mesotheliomas. This method for producing consistent, reproducible 3D spheroids may allow for improved testing of novel monoclonal antibodies and other agents for their ability to penetrate solid tumors for cancer therapy.

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Section: Introductionmentioning

confidence: 99%

Rapid Generation of In Vitro Multicellular Spheroids for the Study of Monoclonal Antibody Therapy

Phung¹,

Barbone²,

Broaddus³

et al. 2011

J. Cancer

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“…2 ). A similar behavior described by the Gompertz function characterizes tumor growth in vivo [13]. The deceleration in spheroid growth can be attributed to the change in the ratio between different cell populations as its size rises and to the increasingly hindered supply of oxygen and nutrients deep into the spheroid: the increase in the proportion of non-dividing (resting) cells and/or death of resting cells accompanied by enlargement of the necrotic core [3, 4].…”

Section: Resultsmentioning

confidence: 93%

Spheroids of HER2-Positive Breast Adenocarcinoma for Studying Anticancer Immunotoxins In Vitro

Balalaeva¹,

Соколова²,

Puzhikhina³

et al. 2017

Acta Naturae

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Tumor response to therapeutic treatment is largely determined by its heterogeneity and the presence of intercellular junctions, hindering the penetration of large molecules deep into the three-dimensional structure of the tumor. In that context, 3D in vitro tumor models such as cancer cell spheroids are becoming increasingly popular. We obtained spheroids of human breast adenocarcinoma SKBR-3 overexpressing the HER2 cancer marker. The toxicity of HER2-targeted immunotoxin 4D5scFv-PE40 against spheroids was shown to be several orders of magnitude lower compared to a monolayer cell culture. The significant difference in the severity of the immunotoxin effect can be explained by the fact that it ineffectively penetrates the spheroid and predominantly influences the cells of the outer layers. The resulting tumor spheroid model can be used in development of drugs for targeted therapy as well as to study ways to improve the efficiency of anticancer agents by targeting cell–cell contacts.

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“…There is disagreement concerning the penetration of MAbs into tumour spheroids. Both specific and non-specific MAbs have been reported to penetrate into the interior of spheroids within 6 to 11 h (McFadden and Kwok, 1988;Hjelstuen et al, 1996). In contrast, it has been reported that intact MAbs targeted to cell membrane antigens bind only to the outer 1-3 cell layers of the spheroid, whereas MAb fragments penetrate 8-10 cell layers (Mairs et al, 1991).…”

Section: Discussionmentioning

confidence: 99%

Cytotoxicity of α-particle-emitting astatine-211-labelled antibody in tumour spheroids: no effect of hyperthermia

Hauck

Larsen²,

Welsh³

et al. 1998

Br J Cancer

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Summary The high linear energy transfer, a-particle-emitting radionuclide astatine-211 (21'At) is of interest for certain therapeutic applications; however, because of the 55-to 70-jim path length of its a-particles, achieving homogeneous tracer distribution is critical. Hyperthermia may enhance the therapeutic efficacy of a-particle endoradiotherapy if it can improve tracer distribution. In this study, we have investigated whether hyperthermia increased the cytotoxicity of an 21'At-labelled monoclonal antibody (MAb) in tumour spheroids with a radius (approximately 100 gm) greater than the range of 21'At a-particles. Hyperthermia for 1 h at 420C was used because this treatment itself resulted in no regrowth delay. Radiolabelled chimeric MAb 81C6 reactive with the extracellular matrix antigen tenascin was added to spheroids grown from the D-247 MG human glioma cell line at activity concentrations ranging from 0.125 to 250 kBq ml-'. A significant regrowth delay was observed at 125 and 250 kBq ml-' in both hyperthermia-treated and untreated spheroids. For groups receiving hyperthermia, no increase in cytotoxicity was seen compared with normothermic controls at any activity concentration. These results and those from autoradiographs indicate that hyperthermia at 420C for 1 h had no significant effect on the uptake or distribution of this antitenascin MAb in D-247 MG spheroids.Keywords: monoclonal antibody; hyperthermia; spheroid; astatine-21 1; a-emitterThe use of a-particle-emitting radionuclides for endoradiotherapy has several potential advantages compared with 1-emitters. The high linear energy transfer (LET) of a-particles makes them highly cytotoxic, rendering even hypoxic cells vulnerable. In addition, a-particles have relatively short effective path lengths in tissue, decreasing the radiation delivered to normal tissues located in close proximity to the tumour. For example, the mean path length of the a-particles emitted by 7.2-h half-life 21'At is 55-70 gm compared with 800 gm for the 5-particles emitted by '3'I (Gaze et al, 1992;. In contrast, the short a-particle range is potentially problematic because of the necessity for achieving homogeneous distribution of the radionuclide in the tumour for effective therapy. The limited range of a-particles has led investigators to focus therapeutic applications in which the tumour is present in thin sheets, such as neoplastic meningitis and peritoneal metastasis. The efficacy of 21'At-labelled monoclonal antibodies (MAbs) in a rat model of neoplastic meningitis has been encouraging .Micrometastatic disease could offer another potential application of a-emitters if sufficiently homogeneous tracer distribution could be achieved. The determination of the most appropriate radionuclide for the treatment of small volume disease has been the focus of both experimental and theoretical studies (Humm and Cobb, 1990;Langmuir et al, 1990; 1992a higher absorbed dose fraction within the tumour compared with long-range 3-emitters such as 90Y, particularly for lesions of less than...

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Penetration and Binding of Monoclonal Antibody in Human Osteosarcoma Multicell Spheroids: Comparison of confocal laser scanning microscopy and autoradiography

Cited by 16 publications

References 20 publications

Rapid Generation of In Vitro Multicellular Spheroids for the Study of Monoclonal Antibody Therapy

Rapid Generation of In Vitro Multicellular Spheroids for the Study of Monoclonal Antibody Therapy

Spheroids of HER2-Positive Breast Adenocarcinoma for Studying Anticancer Immunotoxins In Vitro

Cytotoxicity of α-particle-emitting astatine-211-labelled antibody in tumour spheroids: no effect of hyperthermia

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