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Gene therapy of anaplastic thyroid carcinoma with a single-chain interleukin-12 fusion protein.

Abstract

Anaplastic thyroid carcinoma is the most aggressive type of thyroid malignancy with a mean survival time of less than 8 months. No effective therapeutic approach is currently available, making the development of novel treatments necessary. Interleukin (IL)-12 is a proinflammatory heterodimeric cytokine with strong antitumor activity. In the present study, we investigated the potential of IL-12 gene therapy for anaplastic thyroid carcinoma in BALB/c (nu/nu) nude mice. A single-chain IL-12 fusion protein construct was created to assure equal expression of its p35 and p40 subunits. Human anaplastic thyroid carcinoma cell line ARO was stably transfected with an IL-12 expression plasmid under the control of cytomegalovirus (CMV) promoter (scIL-12/CMVpDNA). High levels of functional IL-12 (26.78 +/- 4.11 ng/ml per 10(6) cells per 48 hr) were produced by scIL-12-transfected ARO cells (ARO/IL-12). Tumorigenicity in nude mice was completely lost in scIL-12-transfected ARO cells, as demonstrated by the lack of tumor formation after subcutaneous injection of 2 x 10(6) ARO/IL-12 cells, even though there was no difference in cell proliferation between ARO and ARO/IL-12 cells. Tumor growth was observed after challenge with ARO tumor cells, indicating that protective immunity had not developed against the parental cells. Furthermore, the growth rate of established subcutaneous ARO tumors was significantly reduced by either subcutaneous injection of 2 x 10(6) ARO/IL-12 cells weekly or intramuscular injection of 50 microg scIL-12/CMVpDNA twice weekly. The antineoplastic activity of ARO/IL-12 cells was, however, abrogated by intraperitoneal injection of anti-natural killer (NK) cell antibody. Moreover, significantly higher number of ARO/IL-12 cells and ARO cells were killed by splenocytes from nude mice previously treated with ARO/IL-12 compared to those treated with ARO cells (32% vs. 9% when ARO were used as target cells, 43% vs. 17% when ARO/IL12 were used as target cells; p < 0.01) in an in vitro cytotoxicity assay. Again, tumor cell killing was neutralized by the addition of anti-NK cell antibody in the assay. In conclusion, we have demonstrated successful gene therapy with a scIL-12 fusion protein against anaplastic thyroid carcinoma in an in vivo model. The immune response against ARO/IL-12 cells is mediated by NK cells. These results may set the stage for clinical application of IL-12 gene therapy for poorly differentiated thyroid carcinoma.

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  • Authors+Show Affiliations

    ,

    Department of Genetics, King Faisal Specialist Hospital and Research Center, Riyadh 11211, Saudi Arabia. yufei@kfshrc.edu.sa

    , , , , , ,

    Source

    Human gene therapy 14:18 2003 Dec 10 pg 1741-51

    MeSH

    Angiogenesis Inhibitors
    Animals
    Carcinoma
    Cell Line, Tumor
    Cytomegalovirus
    Female
    Genetic Therapy
    Genetic Vectors
    Interleukin-12
    Killer Cells, Natural
    Mice
    Mice, Inbred BALB C
    Mice, Nude
    Neoplasms, Experimental
    Plasmids
    Thyroid Neoplasms
    Transfection

    Pub Type(s)

    Journal Article

    Language

    eng

    PubMed ID

    14670125

    Citation

    Shi, Yufei, et al. "Gene Therapy of Anaplastic Thyroid Carcinoma With a Single-chain Interleukin-12 Fusion Protein." Human Gene Therapy, vol. 14, no. 18, 2003, pp. 1741-51.
    Shi Y, Parhar RS, Zou M, et al. Gene therapy of anaplastic thyroid carcinoma with a single-chain interleukin-12 fusion protein. Hum Gene Ther. 2003;14(18):1741-51.
    Shi, Y., Parhar, R. S., Zou, M., Baitei, E., Kessie, G., Farid, N. R., ... Al-Mohanna, F. A. (2003). Gene therapy of anaplastic thyroid carcinoma with a single-chain interleukin-12 fusion protein. Human Gene Therapy, 14(18), pp. 1741-51.
    Shi Y, et al. Gene Therapy of Anaplastic Thyroid Carcinoma With a Single-chain Interleukin-12 Fusion Protein. Hum Gene Ther. 2003 Dec 10;14(18):1741-51. PubMed PMID: 14670125.
    * Article titles in AMA citation format should be in sentence-case
    TY - JOUR T1 - Gene therapy of anaplastic thyroid carcinoma with a single-chain interleukin-12 fusion protein. AU - Shi,Yufei, AU - Parhar,Ranjit S, AU - Zou,Minjing, AU - Baitei,Essa, AU - Kessie,George, AU - Farid,Nadir R, AU - Alzahrani,Ali, AU - Al-Mohanna,Futwan A, PY - 2003/12/13/pubmed PY - 2004/7/14/medline PY - 2003/12/13/entrez SP - 1741 EP - 51 JF - Human gene therapy JO - Hum. Gene Ther. VL - 14 IS - 18 N2 - Anaplastic thyroid carcinoma is the most aggressive type of thyroid malignancy with a mean survival time of less than 8 months. No effective therapeutic approach is currently available, making the development of novel treatments necessary. Interleukin (IL)-12 is a proinflammatory heterodimeric cytokine with strong antitumor activity. In the present study, we investigated the potential of IL-12 gene therapy for anaplastic thyroid carcinoma in BALB/c (nu/nu) nude mice. A single-chain IL-12 fusion protein construct was created to assure equal expression of its p35 and p40 subunits. Human anaplastic thyroid carcinoma cell line ARO was stably transfected with an IL-12 expression plasmid under the control of cytomegalovirus (CMV) promoter (scIL-12/CMVpDNA). High levels of functional IL-12 (26.78 +/- 4.11 ng/ml per 10(6) cells per 48 hr) were produced by scIL-12-transfected ARO cells (ARO/IL-12). Tumorigenicity in nude mice was completely lost in scIL-12-transfected ARO cells, as demonstrated by the lack of tumor formation after subcutaneous injection of 2 x 10(6) ARO/IL-12 cells, even though there was no difference in cell proliferation between ARO and ARO/IL-12 cells. Tumor growth was observed after challenge with ARO tumor cells, indicating that protective immunity had not developed against the parental cells. Furthermore, the growth rate of established subcutaneous ARO tumors was significantly reduced by either subcutaneous injection of 2 x 10(6) ARO/IL-12 cells weekly or intramuscular injection of 50 microg scIL-12/CMVpDNA twice weekly. The antineoplastic activity of ARO/IL-12 cells was, however, abrogated by intraperitoneal injection of anti-natural killer (NK) cell antibody. Moreover, significantly higher number of ARO/IL-12 cells and ARO cells were killed by splenocytes from nude mice previously treated with ARO/IL-12 compared to those treated with ARO cells (32% vs. 9% when ARO were used as target cells, 43% vs. 17% when ARO/IL12 were used as target cells; p < 0.01) in an in vitro cytotoxicity assay. Again, tumor cell killing was neutralized by the addition of anti-NK cell antibody in the assay. In conclusion, we have demonstrated successful gene therapy with a scIL-12 fusion protein against anaplastic thyroid carcinoma in an in vivo model. The immune response against ARO/IL-12 cells is mediated by NK cells. These results may set the stage for clinical application of IL-12 gene therapy for poorly differentiated thyroid carcinoma. SN - 1043-0342 UR - https://www.unboundmedicine.com/medline/citation/14670125/Gene_therapy_of_anaplastic_thyroid_carcinoma_with_a_single_chain_interleukin_12_fusion_protein_ L2 - https://www.liebertpub.com/doi/full/10.1089/104303403322611755?url_ver=Z39.88-2003&amp;rfr_id=ori:rid:crossref.org&amp;rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -