Hemangiosarcoma and its cancer stem cell subpopulation are effectively killed by a toxin targeted through epidermal growth factor and urokinase receptors

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Abstract

Targeted toxins have the potential to overcome intrinsic or acquired resistance of cancer cells to conventional cytotoxic agents. Here, we hypothesized that EGFuPA-toxin, a bispecific ligand-targeted toxin (BLT) consisting of a deimmunized Pseudomonas exotoxin (PE) conjugated to epidermal growth factor and urokinase, would efficiently target and kill cells derived from canine hemangiosarcoma (HSA), a highly chemotherapy resistant tumor, as well as cultured hemangiospheres, used as a surrogate for cancer stem cells (CSC). EGFuPA-toxin showed cytotoxicity in four HSA cell lines (Emma, Frog, DD-1 and SB) at a concentration of ≤100 nM, and the cytotoxicity was dependent on specific ligand-receptor interactions. Monospecific targeted toxins also killed these chemoresistant cells; in this case, a "threshold" level of EGFR expression appeared to be required to make cells sensitive to the monospecific EGF-toxin, but not to the monospecific uPA-toxin. The IC50 of CSCs was higher by approximately two orders of magnitude as compared to non-CSCs, but these cells were still sensitive to EGFuPA-toxin at nanomolar (i.e., pharmacologically relevant) concentrations, and when targeted by EGFuPA-toxin, resulted in death of the entire cell population. Taken together, our results support the use of these toxins to treat chemoresistant tumors such as sarcomas, including those that conform to the CSC model. Our results also support the use of companion animals with cancer for further translational development of these cytotoxic molecules. What's new? Sarcomas are aggressive, highly chemoresistant tumors with few treatment options. These data show for the first time that a bispecific ligand-targeted toxin composed of a Pseudomonas exotoxin conjugated to epidermal growth factor and urokinase (EGFuPA-toxin) induces cytotoxicity of highly chemoresistant sarcoma cells, including a subpopulation of cancer stem cells. The findings, which were obtained using cells derived from canine hemangiosarcoma, support the use of companion animal tumors for translational development of EGFuPA-toxin. While sarcomas are infrequent in humans, they occur spontaneously and frequently in domestic dogs, so the use of canine tumors could help accelerate further clinical development in humans.

Original languageEnglish (US)
Pages (from-to)1936-1944
Number of pages9
JournalInternational Journal of Cancer
Volume133
Issue number8
DOIs
StatePublished - Oct 15 2013

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Immunotoxins
Hemangiosarcoma
Neoplastic Stem Cells
Urokinase-Type Plasminogen Activator
Sarcoma
Neoplasms
Epidermal Growth Factor
Canidae
Exotoxins
Pets
Ligands
Pseudomonas
Cytotoxins
Human Development
ErbB Receptors
Anura
Inhibitory Concentration 50
Cell Death
Dogs
Drug Therapy

Keywords

  • cancer stem cell
  • epidermal growth factor
  • hemangiosarcoma
  • targeted therapies
  • urokinase

Cite this

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title = "Hemangiosarcoma and its cancer stem cell subpopulation are effectively killed by a toxin targeted through epidermal growth factor and urokinase receptors",
abstract = "Targeted toxins have the potential to overcome intrinsic or acquired resistance of cancer cells to conventional cytotoxic agents. Here, we hypothesized that EGFuPA-toxin, a bispecific ligand-targeted toxin (BLT) consisting of a deimmunized Pseudomonas exotoxin (PE) conjugated to epidermal growth factor and urokinase, would efficiently target and kill cells derived from canine hemangiosarcoma (HSA), a highly chemotherapy resistant tumor, as well as cultured hemangiospheres, used as a surrogate for cancer stem cells (CSC). EGFuPA-toxin showed cytotoxicity in four HSA cell lines (Emma, Frog, DD-1 and SB) at a concentration of ≤100 nM, and the cytotoxicity was dependent on specific ligand-receptor interactions. Monospecific targeted toxins also killed these chemoresistant cells; in this case, a {"}threshold{"} level of EGFR expression appeared to be required to make cells sensitive to the monospecific EGF-toxin, but not to the monospecific uPA-toxin. The IC50 of CSCs was higher by approximately two orders of magnitude as compared to non-CSCs, but these cells were still sensitive to EGFuPA-toxin at nanomolar (i.e., pharmacologically relevant) concentrations, and when targeted by EGFuPA-toxin, resulted in death of the entire cell population. Taken together, our results support the use of these toxins to treat chemoresistant tumors such as sarcomas, including those that conform to the CSC model. Our results also support the use of companion animals with cancer for further translational development of these cytotoxic molecules. What's new? Sarcomas are aggressive, highly chemoresistant tumors with few treatment options. These data show for the first time that a bispecific ligand-targeted toxin composed of a Pseudomonas exotoxin conjugated to epidermal growth factor and urokinase (EGFuPA-toxin) induces cytotoxicity of highly chemoresistant sarcoma cells, including a subpopulation of cancer stem cells. The findings, which were obtained using cells derived from canine hemangiosarcoma, support the use of companion animal tumors for translational development of EGFuPA-toxin. While sarcomas are infrequent in humans, they occur spontaneously and frequently in domestic dogs, so the use of canine tumors could help accelerate further clinical development in humans.",
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author = "Schappa, {Jill T.} and Frantz, {Aric M.} and Gorden, {Brandi H.} and Dickerson, {Erin B.} and Vallera, {Daniel A.} and Modiano, {Jaime F.}",
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T1 - Hemangiosarcoma and its cancer stem cell subpopulation are effectively killed by a toxin targeted through epidermal growth factor and urokinase receptors

AU - Schappa, Jill T.

AU - Frantz, Aric M.

AU - Gorden, Brandi H.

AU - Dickerson, Erin B.

AU - Vallera, Daniel A.

AU - Modiano, Jaime F.

PY - 2013/10/15

Y1 - 2013/10/15

N2 - Targeted toxins have the potential to overcome intrinsic or acquired resistance of cancer cells to conventional cytotoxic agents. Here, we hypothesized that EGFuPA-toxin, a bispecific ligand-targeted toxin (BLT) consisting of a deimmunized Pseudomonas exotoxin (PE) conjugated to epidermal growth factor and urokinase, would efficiently target and kill cells derived from canine hemangiosarcoma (HSA), a highly chemotherapy resistant tumor, as well as cultured hemangiospheres, used as a surrogate for cancer stem cells (CSC). EGFuPA-toxin showed cytotoxicity in four HSA cell lines (Emma, Frog, DD-1 and SB) at a concentration of ≤100 nM, and the cytotoxicity was dependent on specific ligand-receptor interactions. Monospecific targeted toxins also killed these chemoresistant cells; in this case, a "threshold" level of EGFR expression appeared to be required to make cells sensitive to the monospecific EGF-toxin, but not to the monospecific uPA-toxin. The IC50 of CSCs was higher by approximately two orders of magnitude as compared to non-CSCs, but these cells were still sensitive to EGFuPA-toxin at nanomolar (i.e., pharmacologically relevant) concentrations, and when targeted by EGFuPA-toxin, resulted in death of the entire cell population. Taken together, our results support the use of these toxins to treat chemoresistant tumors such as sarcomas, including those that conform to the CSC model. Our results also support the use of companion animals with cancer for further translational development of these cytotoxic molecules. What's new? Sarcomas are aggressive, highly chemoresistant tumors with few treatment options. These data show for the first time that a bispecific ligand-targeted toxin composed of a Pseudomonas exotoxin conjugated to epidermal growth factor and urokinase (EGFuPA-toxin) induces cytotoxicity of highly chemoresistant sarcoma cells, including a subpopulation of cancer stem cells. The findings, which were obtained using cells derived from canine hemangiosarcoma, support the use of companion animal tumors for translational development of EGFuPA-toxin. While sarcomas are infrequent in humans, they occur spontaneously and frequently in domestic dogs, so the use of canine tumors could help accelerate further clinical development in humans.

AB - Targeted toxins have the potential to overcome intrinsic or acquired resistance of cancer cells to conventional cytotoxic agents. Here, we hypothesized that EGFuPA-toxin, a bispecific ligand-targeted toxin (BLT) consisting of a deimmunized Pseudomonas exotoxin (PE) conjugated to epidermal growth factor and urokinase, would efficiently target and kill cells derived from canine hemangiosarcoma (HSA), a highly chemotherapy resistant tumor, as well as cultured hemangiospheres, used as a surrogate for cancer stem cells (CSC). EGFuPA-toxin showed cytotoxicity in four HSA cell lines (Emma, Frog, DD-1 and SB) at a concentration of ≤100 nM, and the cytotoxicity was dependent on specific ligand-receptor interactions. Monospecific targeted toxins also killed these chemoresistant cells; in this case, a "threshold" level of EGFR expression appeared to be required to make cells sensitive to the monospecific EGF-toxin, but not to the monospecific uPA-toxin. The IC50 of CSCs was higher by approximately two orders of magnitude as compared to non-CSCs, but these cells were still sensitive to EGFuPA-toxin at nanomolar (i.e., pharmacologically relevant) concentrations, and when targeted by EGFuPA-toxin, resulted in death of the entire cell population. Taken together, our results support the use of these toxins to treat chemoresistant tumors such as sarcomas, including those that conform to the CSC model. Our results also support the use of companion animals with cancer for further translational development of these cytotoxic molecules. What's new? Sarcomas are aggressive, highly chemoresistant tumors with few treatment options. These data show for the first time that a bispecific ligand-targeted toxin composed of a Pseudomonas exotoxin conjugated to epidermal growth factor and urokinase (EGFuPA-toxin) induces cytotoxicity of highly chemoresistant sarcoma cells, including a subpopulation of cancer stem cells. The findings, which were obtained using cells derived from canine hemangiosarcoma, support the use of companion animal tumors for translational development of EGFuPA-toxin. While sarcomas are infrequent in humans, they occur spontaneously and frequently in domestic dogs, so the use of canine tumors could help accelerate further clinical development in humans.

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KW - epidermal growth factor

KW - hemangiosarcoma

KW - targeted therapies

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