Antitumor activity in mice of an immunotoxin made with anti-transferrin receptor and a recombinant form of Pseudomonas exotoxin.

Batra, Janendra K.; Jinno, Yoshihiro; Chaudhary, Vijay; Kondo, Toshihiko; FitzGerald, David J.; Pastan, Ira

doi:10.1073/pnas.86.21.8545

Cited by 67 publications

(54 citation statements)

References 17 publications

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“…The use of Tf-adriamycin in a preliminary clinical study in acute leukemia patients has been reported [17]. A promising further application in vivo has been reported using the immunotoxin antiTfR-LysPE40, where intraperitoneal administration caused regression of subcutaneous A431 tumors [54].…”

Section: Targeting To Various Cell Types Via the Transferrin Receptormentioning

confidence: 99%

“…Moreover, the Tf-conjugate, unlike native DT, was also highly toxic to murine cells. Pastan and co-workers [54] constructed the targetcell-specific immunotoxin antiTfR-LysPE40 by conjugation of antiTfR antibodies with recombinant 40-kDa fragments of Pseudomonas exotoxin that lack its cell-binding domain.…”

Section: Tf and Tfr Conjugates For Improved Uptake Into Cellsmentioning

confidence: 99%

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Delivery of drugs, proteins and genes into cells using transferrin as a ligand for receptor-mediated endocytosis

Wagner

Curiel

Cotten

1994

Advanced Drug Delivery Reviews

259

128

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Section: Targeting To Various Cell Types Via the Transferrin Receptormentioning

confidence: 99%

Section: Tf and Tfr Conjugates For Improved Uptake Into Cellsmentioning

confidence: 99%

Delivery of drugs, proteins and genes into cells using transferrin as a ligand for receptor-mediated endocytosis

Wagner

Curiel

Cotten

1994

Advanced Drug Delivery Reviews

259

128

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“…[17][18][19][20]48,49 Given this background, we tested our hypothesis that a similar targeting strategy might be successful when directed to rapidly proliferating but nontransformed smooth muscle cells. To this end, we determined whether TGFa-PE40, a chimeric toxin previously found to be extremely potent in killing certain cancer cells,20 would be cytotoxic to rapidly proliferating smooth muscle cells.…”

Section: Discussionmentioning

confidence: 99%

“…'1-14 This characteristic of certain transformed cells stimulated the development of cytotoxic agents specifically targeted to the cancer cells that overexpress these receptors. [15][16][17][18][19][20] We speculated that because activated smooth muscle cells also express higher numbers of certain cell surface receptors than do nonactivated cells, their excessive growth might be controlled by an analogous therapeutic approach.…”

mentioning

confidence: 99%

Cytotoxic effects of a recombinant chimeric toxin on rapidly proliferating vascular smooth muscle cells.

et al. 1991

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BACKGROUND Restenosis after percutaneous transluminal coronary angioplasty is associated with activation of medial smooth muscle cells (SMCs); they proliferate, migrate to the subintima, and narrow the vessel lumen. Cancer cells often express more cell surface receptors than do normal cells. This has allowed tumor cells to be specifically targeted using cytotoxic agents. We have examined whether a similar concept can be applied to rapidly proliferating but nontransformed SMCs. Pseudomonas exotoxin (PE; MW, 66 kDa) is a potent toxin that kills cells by inhibiting protein synthesis; its toxicity is diminished when its cell recognition domain is deleted to produce a 40-kDa protein (PE40). METHODS AND RESULTS A complementary DNA encoding transforming growth factor alpha (TGF alpha) was ligated to that encoding PE40 and the chimeric toxin TGF alpha-PE40, which is cytotoxic to cancer cells displaying epidermal growth factor (EGF) receptors, was expressed in Escherichia coli. The ability of this toxin to kill proliferating SMCs was tested. When cells were seeded at low density (2,500 cells/cm2) and grown in medium supplemented with 10% fetal bovine serum, they were found to be rapidly proliferating; these cells were very sensitive to the cytotoxic effects of TGF alpha-PE40 (ID50, 4.0 +/- 0.17 ng/ml). In contrast, cytotoxicity was 30-fold less (ID50, 125 +/- 23 ng/ml; p less than 0.0004) when cells were in a quiescent state (grown in medium supplemented with 0.5% fetal bovine serum). CONCLUSIONS Competition studies using excess EGF indicated that the cytotoxic effects of TGF alpha-PE40 are specifically mediated by the EGF receptor. EGF receptor binding analysis demonstrated that rapidly proliferating SMCs display 10-fold more EGF receptors than do quiescent SMCs in vitro. Thus, a chimeric toxin targeted toward the EGF receptor can selectively kill rapidly proliferating SMCs. Whether this toxin or other chimeric toxins directed against other cell surface receptors will effectively inhibit SMCs proliferating in vivo or be useful in preventing restenosis remains to be determined.

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“…528) was generously provided by J. Mendelson (Masui et al 1984}. A conjugate (anti-EGFR-PE) of this antibody with Pseudomonas exotoxin (as a hapten) was the generous gift of I. Pastan and J. Batra (Batra et al 1989). The labeling sequence was as follows: anti-EGFR-PE at 1 ~g/ml, rabbit anti-Pseudomonas exotoxin (anti-PE) (the generous gift of D. FitzGerald and I. Pastan) at 2 ~g/ml, and goat anti-rabbit immunoglobulin G conjugated with either horseradish peroxidase (5 ~g/ml; Jackson ImmunoResearch) or 1 nm of colloidal gold (Janssen Auroprobe; 1 : 100 dilution); labeling was done at room temperature for 60 min.…”

Section: Histology and Immunocytochemistrymentioning

confidence: 99%

Inactivation of a sperm motility gene by insertion of an epidermal growth factor receptor transgene whose product is overexpressed and compartmentalized during spermatogenesis.

Merlino¹,

Stahle²,

Jhappan³

et al. 1991

Genes Dev.

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Transgenic mice were generated with a human epidermal growth factor (EGF) receptor cDNA driven by the chicken 13-actin gene promoter. One line (AE24) that exhibited a unique expression pattern in which dramatically elevated levels of EGF receptor RNA were found only in the testis was established, suggesting that the 13-actin promoter was being influenced by an adjacent testis-specific enhancer. EGF receptor RNA was detected in primary spermatocytes, whereas the synthesis of receptor protein was restricted to elongate spermatids, indicating that transgene expression was under translational control. At spermiation, the EGF receptor was sequestered in residual bodies and excluded from mature sperm by a compartmentalization mechanism. About half of AE24 homozygous males were sterile because of sperm paralysis, whereas heterozygous males and females of either genotype were completely fertile. Electron microscopic analysis of sperm flagella from sterile AE24 homozygotes revealed an aberrant axonemal structure in which outer doublet microtubules were missing from the middle piece, resembling changes observed in the sperm of some infertile humans. Flagellar axonemal disassembly was observed in the vas deferens and epididymis but not in the testis, suggesting that outer doublets were assembled in a grossly normal manner but possessed a latent instability. These results demonstrate that in the AE24 mouse line the EGF receptor transgene was integrated into and inactivated an endogenous autosomal gene, causing sperm flagellar axonemal disruption and male sterility.

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Antitumor activity in mice of an immunotoxin made with anti-transferrin receptor and a recombinant form of Pseudomonas exotoxin.

Cited by 67 publications

References 17 publications

Delivery of drugs, proteins and genes into cells using transferrin as a ligand for receptor-mediated endocytosis

Delivery of drugs, proteins and genes into cells using transferrin as a ligand for receptor-mediated endocytosis

Cytotoxic effects of a recombinant chimeric toxin on rapidly proliferating vascular smooth muscle cells.

Inactivation of a sperm motility gene by insertion of an epidermal growth factor receptor transgene whose product is overexpressed and compartmentalized during spermatogenesis.

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