The diphtheria toxin/urokinase fusion protein (DTAT) is selectively toxic to CD87 expressing leukemic cells

Ramage, J.; Vallera, Daniel A.; Black, J; Aplan, Peter D.; Kees, Ursula R.; Frankel, Arthur E.

doi:10.1016/s0145-2126(02)00077-2

Cited by 35 publications

(29 citation statements)

References 44 publications

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“…Thus, our first AML fusion toxins (DT 388 GMCSF, DT 388 IL3, and DTAT) were potentially toxic to normal tissues bearing their respective receptors-GM-CSFR, interleukin 3 receptor (IL-3R), or uPAR. 4,12,16 Clinical testing of the first AML fusion toxin DT 388 GMCSF confirmed clinical efficacy but with associated damage to GM-CSFR-containing normal cells. This led to significant liver injury.…”

Section: Discussionmentioning

confidence: 97%

“…The uPA/uPAR system is overexpressed in a variety of tumors including leukemias. 10,11 AML blasts overexpress uPA and uPAR, 12 whereas most normal tissues have no or little uPA and uPAR expression, except when they are up-regulated during certain physiologic processes such as wound healing and extracellular matrix remodeling. 10,11 Consequently, we chose to enhance DT 388 GMCSF specificity to AML by changing its furin cleavage region ( 163 RVRRSV 170 ) to a uPA cleavage sequence ( 163 GSGRSA 170 ), yielding DTU2GMCSF.…”

Section: Introductionmentioning

confidence: 99%

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A urokinase-activated recombinant diphtheria toxin targeting the granulocyte-macrophage colony-stimulating factor receptor is selectively cytotoxic to human acute myeloid leukemia blasts

Abi-Habib

Liu

Bugge

et al. 2004

Blood

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Novel agents to treat acute myeloid leukemia (AML) are needed with increased efficacy and specificity. We have synthesized a dual-specificity fusion toxin DTU2GMCSF composed of the catalytic and translocation domains of diphtheria toxin (DT) fused to the granulocytemacrophage colony-stimulating factor (GM-CSF) in which the DT furin cleavage site 163 RVRRSV 170 is modified to a urokinase plasminogen activator (uPA) cleavage site 163 GSGRSA 170 , termed U2. DTU2GMCSF was highly toxic to the TF1-vRaf AML cell line (proliferation inhibition assay; IC 50 ‫؍‬ 3.14 pM), and this toxicity was greatly inhibited following pretreatment with anti-uPA and anti-GM-CSF antibodies. The activity of this toxin was then tested on a larger group of 13 human AML cell lines; 5 of the 13 cell lines were sensitive to DTU2GMCSF. An additional 5 of the 13 cell lines became sensitive when exogenous pro-uPA was added. Sensitivity to DTU2GMCSF strongly correlated with the expression levels of uPA receptors (uPARs) and GM-CSF receptors (GMCSFRs) as well as with total uPA levels. DTU2GMCSF was less toxic to normal cells expressing uPAR or GMCSFR alone, that is, human umbilical vein endothelial cells and peripheral macrophages, respectively. These results indicate that DTU2GMCSF may be a selective and potent agent for the treatment of patients with

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Section: Discussionmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

A urokinase-activated recombinant diphtheria toxin targeting the granulocyte-macrophage colony-stimulating factor receptor is selectively cytotoxic to human acute myeloid leukemia blasts

Abi-Habib

Liu

Bugge

et al. 2004

Blood

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“…It is therefore judicious to test for the expression these new therapeutic targets on blast cells from patients liable to enter clinical trials using or evaluating such drugs. 60 The list of useful markers to test at diagnosis can therefore be extended, according to current schedules applied in a given center, to include the well-known targets of rituximab (CD20), 61 or alemtuzumab (CD52), 62 but also CD45, 63 CD33, 64 CD116, 65 CD123, 55 CD44, 66 CD22 67 or CD87 68 as a non-exhaustive list.…”

Section: Useful Additional Markersmentioning

confidence: 99%

Immunophenotyping of acute leukemia and lymphoproliferative disorders: a consensus proposal of the European LeukemiaNet Work Package 10

et al. 2011

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The European LeukemiaNet (ELN), workpackage 10 (WP10) was designed to deal with diagnosis matters using morphology and immunophenotyping. This group aimed at establishing a consensus on the required reagents for proper immunophenotyping of acute leukemia and lymphoproliferative disorders. Animated discussions within WP10, together with the application of the Delphi method of proposals circulation, quickly led to post-consensual immunophenotyping panels for disorders on the ELN website. In this report, we established a comprehensive description of these panels, both mandatory and complementary, for both types of clinical conditions. The reason for using each marker, sustained by relevant literature information, is provided in detail. With the constant development of immunophenotyping techniques in flow cytometry and related software, this work aims at providing useful guidelines to perform the most pertinent exploration at diagnosis and for follow-up, with the best cost benefit in diseases, the treatment of which has a strong impact on health systems.

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“…Deciphering the missing molecular details of diphtheria toxin's cellular entry is relevant for understanding the entry of other toxins [2] and for the development of biomedical applications of targeted drug delivery. Indeed, diphtheria toxin has already been utilized as a prospective anti-cancer agent for the targeted delivery of cytotoxic therapy to cancer cells [3][4][5][6][7][8][9][10][11][12][13][14][15][16]. Normally, the targeted delivery is achieved by deleting the cell receptor-binding R-domain and combining the remaining portion (containing T-and C-domains) with proteins that selectively bind to the surface of cancer cells.…”

Section: Introductionmentioning

confidence: 99%

Cellular Entry of Diphtheria Toxin Does Not Require Formation of the Open-Channel State by its Translocation Domain

Rodnin¹,

Vargas-Uribe²,

Ladokhin³

2018

Biophysical Journal

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Cellular entry of diphtheria toxin is a multistage process involving receptor targeting, endocytosis, and translocation of the catalytic domain across the endosomal membrane into the cytosol. The latter is ensured by the translocation (T) domain of the toxin, capable of undergoing conformational refolding and membrane insertion in response to the acidification of the endosomal environment. While numerous now classical studies have demonstrated the formation of an ion-conducting conformation-the Open-Channel State (OCS)-as the final step of the refolding pathway, it remains unclear whether this channel constitutes an in vivo translocation pathway or is a byproduct of the translocation. To address this question, we measure functional activity of known OCS-blocking mutants with H-to-Q replacements of C-terminal histidines of the T-domain. We also test the ability of these mutants to translocate their own N-terminus across lipid bilayers of model vesicles. The results of both experiments indicate that translocation activity does not correlate with previously published OCS activity. Finally, we determined the topology of TH5 helix in membrane-inserted T-domain using W281 fluorescence and its depth-dependent quenching by brominated lipids. Our results indicate that while TH5 becomes a transbilayer helix in a wild-type protein, it fails to insert in the case of the OCS-blocking mutant H322Q. We conclude that the formation of the OCS is not necessary for the functional translocation by the T-domain, at least in the histidine-replacement mutants, suggesting that the OCS is unlikely to constitute a translocation pathway for the cellular entry of diphtheria toxin in vivo.

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The diphtheria toxin/urokinase fusion protein (DTAT) is selectively toxic to CD87 expressing leukemic cells

Cited by 35 publications

References 44 publications

A urokinase-activated recombinant diphtheria toxin targeting the granulocyte-macrophage colony-stimulating factor receptor is selectively cytotoxic to human acute myeloid leukemia blasts

A urokinase-activated recombinant diphtheria toxin targeting the granulocyte-macrophage colony-stimulating factor receptor is selectively cytotoxic to human acute myeloid leukemia blasts

Immunophenotyping of acute leukemia and lymphoproliferative disorders: a consensus proposal of the European LeukemiaNet Work Package 10

Cellular Entry of Diphtheria Toxin Does Not Require Formation of the Open-Channel State by its Translocation Domain

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