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. 2012:2012:721873.
doi: 10.1155/2012/721873. Epub 2012 Feb 14.

Methodology for Anti-Gene Anti-IGF-I Therapy of Malignant Tumours

Jerzy Trojan  1 , Yuexin X Pan, Ming X Wei, Adama Ly, Alexander Shevelev, Maciej Bierwagen, Marie-Yvonne Ardourel, Ladislas A Trojan, Alvaro Alvarez, Christian Andres, Maria C Noguera, Ignacio Briceno, Beatriz H Aristizabal, Heliodor Kasprzak, Huynh T Duc, Donald D Anthony
Affiliations

Methodology for Anti-Gene Anti-IGF-I Therapy of Malignant Tumours

Jerzy Trojan et al. Chemother Res Pract. 2012.

Erratum in

  • Chemother Res Pract. 2012;2012:765703

Abstract

The aim of this study was to establish the criteria for methodology of cellular "anti-IGF-I" therapy of malignant tumours and particularly for glioblastoma multiforme. The treatment of primary glioblastoma patients using surgery, radiotherapy, and chemotherapy was followed by subcutaneous injection of autologous cancer cells transfected by IGF-I antisense/triple helix expression vectors. The prepared cell "vaccines" should it be in the case of glioblastomas or other tumours, have shown a change of phenotype, the absence of IGF-I protein, and expression of MHC-I and B7. The peripheral blood lymphocytes, PBL cells, removed after each of two successive vaccinations, have demonstrated for all the types of tumour tested an increasing level of CD8(+) and CD8(+)28(+) molecules and a switch from CD8(+)11b(+) to CD8(+)11. All cancer patients were supervised for up to 19 months, the period corresponding to minimum survival of glioblastoma patients. The obtained results have permitted to specify the common criteria for "anti-IGF-I" strategy: characteristics sine qua non of injected "vaccines" (cloned cells IGF-I(-) and MHC-I(+)) and of PBL cells (CD8(+) increased level).

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Figures

Figure 1
Figure 1
Mechanism of anti-gene anti-IGF-I (antisense/triple helix) therapy of malignant tumours. The case of glioblastoma therapy: hypothetically, the same mechanism should exist in the treatment of other tumours expressing IGF-I. The mechanism of antisense therapy is a combination of an augmentation of the immune antitumour response and of an inhibition of signal transduction pathway that is involved in the transformed phenotype of the tumour. Tumour cells are transfected in vitro with a vector encoding IGF-I cDNA in antisense orientation, or with a vector inducing a formation of triple helix IGF-I structure. The transfected tumor glial cells, in absence of IGF-I, become immunogenic-expressing MHC-I and B7 molecules, and apoptotic as follows. The expression of MHC-I is due to the presence of TAP1; the expression of B7 is related directly with signal transduction pathway: TK/IRS/PI3K/PKC; the phenomenon of apoptosis is also related with signal transduction pathway: TK/IRS/PI3K/AKT/Bcl2 [, –18]. After in vivo injection, together with the antigen presenting cells, APC, they activate the T CD8 (CD8CD28) lymphocytes inducing immune antitumour response against the malignant glioma (expressing MHC-I) [, , –22].
Figure 2
Figure 2
Expression of IGF-I in primary human glioma cell line. RT PCR technique. M—marker. 1—presence of IGF-I in parental non transfected cells (200 bp band of amplified DNA using IGF-I primer; see Methods). 2—absence of IGF-I expression in cells transfected with antisense anti-IGF-I vector. 3—absence of IGF-I expression in cells transfected with triple helix anti-IGF-I vector.
Figure 3
Figure 3
Expression of MHC-I in an established primary human glioma cell line. Flow cytometry analysis (FACScan Becton Dickinson). Panel left: parental nontransfected cells. Panel right: cells transfected with both antisense and triple helix anti-IGF-I vectors.
Figure 4
Figure 4
Examples of in vitro culture of primary human cancer cells derived from surgical biopsies. (a) and (c) Primary ovary cystadenocarcinoma-derived cells. (b) and (d) Primary prostate adenocarcinoma-derived cells. Parental ovary cancer cells (a) and prostate cancer cells (b) are attached efficiently in the fourth day of culture (arrows). “Antisense/triple helix” anti-IGF-I ovary cancer cells (c) and prostate cancer cells (d), both twenty days after transfection, form the established lines characterized often by the clusters of round apoptotic cells, becoming progressively small (c, arrow; d, arrow up). They are accompanied by nonapoptotic and more voluminous cells (d, arrow down) presenting generally elongated shape (c and d). The transfected cells are always different from nontransfected parental cells (a, b), as it was demonstrated previously in cases of human glioma and hepatoma cell lines established from primary tumours of glioblastoma and hepatocarcinoma [30, 31] X400.
Figure 5
Figure 5
Expression of IGF-I in human cancer cell lines. RT PCR technique. M—marker. Lines 1 to 6—presence of IGF-I in parental nontransfected cancer cell lines derived from 1—glioblastoma, 2—hepatocarcinoma, 3—colon adenocarcinoma, 4—ovary cyst adenocarcinoma, 5—uterus endometrial adenocarcinoma, and 6—prostate adenocarcinoma (200 bp band of amplified DNA using IGF-I primer; see Section 2. Lines 7 to 12—absence of IGF-I expression in cancer cells transfected with both antisense and triple helix anti-IGF-I vectors: 7—glioblastoma, 8—hepatocarcinoma, 9—colon adenocarcinoma, 10—ovary cyst adenocarcinoma, 11—uterus endometrial adenocarcinoma, and 12—prostate adenocarcinoma.
Figure 6
Figure 6
Flow cytometric “FACS” peripheral blood lymphocyte CD marker patterns following cellular gene therapy in human cancers. (a) glioblastoma multiforme; (b) hepatocarcinoma; (c) colon adenocarcinoma; (d) ovarian carcinoma; (e) uterine adenocarcinoma; (f) prostate adenocarcinoma. CD molecules were labelled in peripheral blood lymphocytes (PBLs) obtained from prevaccinated and “vaccinated” cancer patients. Each of the first column corresponds to data obtained before vaccinations; each second and third column corresponds to data obtained after one and two successive cellular vaccinations (IGF-I antisense/triple helix cells). Two cases of each of the designated cancers were examined (bar graphs represent the median value of the two cases). Data are expressed as percent of positive cells when compared to the isotype control. Difference in percentage of CD8+ CD11b− and CD8+ CD28+ subpopulations before and after vaccination was strongly significant with a range of P from 0.001 to 0.02 according to the Student's t-test and weakly significant concerning the decreasing CD8+ CD11b+ subpopulation from the relevant patients. The P value for CD8+, CD8+28+, and CD8+11b− (below 0.01) is illustrated in the bar graph for statistical significance. (The original FACS data concerning PBL cells are in the archives of Collegium Medicum of Nicolas Copernic University, Bromberg, Poland; FACS data (n = 504) corresponding to the labeling of CD3, CD4, CD5, CD8, CD8+11b+, CD8+11b−, CD8+28+, CD19, CD3−(16 + 56) + (NK), CD25, CD44, and CD45.)
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References

    1. Stupp R, Hegi ME, Neyns B, et al. Phase I/IIa study of cilengitide and temozolomide with concomitant radiotherapy followed by cilengitide and temozolomide maintenance therapy in patients with newly diagnosed glioblastoma. Journal of Clinical Oncology. 2010;28(16):2712–2718. - PubMed
    1. Wen PY, Yung WKA, Lamborn KR, et al. Phase I/II study of imatinib mesylate for recurrent malignant gliomas: North American Brain Tumor Consortium Study 99-08. Clinical Cancer Research. 2006;12(16):4899–4907. - PubMed
    1. Pan Q, Chanthery Y, Liang WC, et al. Blocking neuropilin-1 function has an additive effect with anti-VEGF to inhibit tumor growth. Cancer Cell. 2007;11(1):53–67. - PubMed
    1. Trojan J, Cloix JF, Ardourel MY, Chatel M, Anthony DD. Insulin-like growth factor type I biology and targeting in malignant gliomas. Neuroscience. 2007;145(3):795–811. - PubMed
    1. Kjaergaard J, Wang LX, Kuriyama H, Shu S, Plautz GE. Active immunotherapy for advanced intracranial murine tumors by using dendritic cell-tumor cell fusion vaccines. Journal of Neurosurgery. 2005;103(1):156–164. - PubMed

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