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Protein Synthesis Inhibition Activity of Mesothelin Targeting Immunotoxin LMB-100 Decreases Concentrations of Oncogenic Signaling Molecules and Secreted Growth Factors

LMB-100 is a mesothelin-targeted recombinant immunotoxin (iTox) that carries a modified Pseuodomonas exotoxin A (PE) payload. PE kills cells by inhibiting synthesis of new proteins. We found that treatment of pancreatic cancer cells with LMB-100 for 24–48 h did not change total protein level despite...

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Autores principales: El-Behaedi, Salma, Landsman, Rebekah, Rudloff, Michael, Kolyvas, Emily, Albalawy, Rakan, Zhang, Xianyu, Bera, Tapan, Collins, Keith, Kozlov, Serguei, Alewine, Christine
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6267581/
https://www.ncbi.nlm.nih.gov/pubmed/30384408
http://dx.doi.org/10.3390/toxins10110447
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author El-Behaedi, Salma
Landsman, Rebekah
Rudloff, Michael
Kolyvas, Emily
Albalawy, Rakan
Zhang, Xianyu
Bera, Tapan
Collins, Keith
Kozlov, Serguei
Alewine, Christine
author_facet El-Behaedi, Salma
Landsman, Rebekah
Rudloff, Michael
Kolyvas, Emily
Albalawy, Rakan
Zhang, Xianyu
Bera, Tapan
Collins, Keith
Kozlov, Serguei
Alewine, Christine
author_sort El-Behaedi, Salma
collection PubMed
description LMB-100 is a mesothelin-targeted recombinant immunotoxin (iTox) that carries a modified Pseuodomonas exotoxin A (PE) payload. PE kills cells by inhibiting synthesis of new proteins. We found that treatment of pancreatic cancer cells with LMB-100 for 24–48 h did not change total protein level despite inducing protein synthesis inhibition (PSI). Further, increased levels of ubiquitinated proteins were detected, indicating that cells may have limited ability to compensate for PSI by reducing protein degradation. Together, these data suggest that PE depletes concentrations of a minority of cellular proteins. We used reverse phase protein array and Luminex assay to characterize this subset. LMB-100 decreased the abundance of 24 of 32 cancer-related proteins (including Bcl-x, Her2, Her3 and MUC16) without compensatory increases in other analytes. Further, cancer cells failed to maintain extracellular concentrations of cancer cell secreted growth factors (CCSGFs), including Vascular Endothelial Growth Factor (VEGF) following treatment with cytostatic LMB-100 doses both in culture and in mouse tumors. Decreased VEGF concentration did not change tumor vasculature density, however, LMB-100 caused tissue-specific changes in concentrations of secreted factors made by non-cancer cells. In summary, our data indicate that PSI caused by cytostatic LMB-100 doses preferentially depletes short-lived proteins such as oncogenic signaling molecules and CCSGFs.
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spelling pubmed-62675812018-12-07 Protein Synthesis Inhibition Activity of Mesothelin Targeting Immunotoxin LMB-100 Decreases Concentrations of Oncogenic Signaling Molecules and Secreted Growth Factors El-Behaedi, Salma Landsman, Rebekah Rudloff, Michael Kolyvas, Emily Albalawy, Rakan Zhang, Xianyu Bera, Tapan Collins, Keith Kozlov, Serguei Alewine, Christine Toxins (Basel) Article LMB-100 is a mesothelin-targeted recombinant immunotoxin (iTox) that carries a modified Pseuodomonas exotoxin A (PE) payload. PE kills cells by inhibiting synthesis of new proteins. We found that treatment of pancreatic cancer cells with LMB-100 for 24–48 h did not change total protein level despite inducing protein synthesis inhibition (PSI). Further, increased levels of ubiquitinated proteins were detected, indicating that cells may have limited ability to compensate for PSI by reducing protein degradation. Together, these data suggest that PE depletes concentrations of a minority of cellular proteins. We used reverse phase protein array and Luminex assay to characterize this subset. LMB-100 decreased the abundance of 24 of 32 cancer-related proteins (including Bcl-x, Her2, Her3 and MUC16) without compensatory increases in other analytes. Further, cancer cells failed to maintain extracellular concentrations of cancer cell secreted growth factors (CCSGFs), including Vascular Endothelial Growth Factor (VEGF) following treatment with cytostatic LMB-100 doses both in culture and in mouse tumors. Decreased VEGF concentration did not change tumor vasculature density, however, LMB-100 caused tissue-specific changes in concentrations of secreted factors made by non-cancer cells. In summary, our data indicate that PSI caused by cytostatic LMB-100 doses preferentially depletes short-lived proteins such as oncogenic signaling molecules and CCSGFs. MDPI 2018-10-31 /pmc/articles/PMC6267581/ /pubmed/30384408 http://dx.doi.org/10.3390/toxins10110447 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
El-Behaedi, Salma
Landsman, Rebekah
Rudloff, Michael
Kolyvas, Emily
Albalawy, Rakan
Zhang, Xianyu
Bera, Tapan
Collins, Keith
Kozlov, Serguei
Alewine, Christine
Protein Synthesis Inhibition Activity of Mesothelin Targeting Immunotoxin LMB-100 Decreases Concentrations of Oncogenic Signaling Molecules and Secreted Growth Factors
title Protein Synthesis Inhibition Activity of Mesothelin Targeting Immunotoxin LMB-100 Decreases Concentrations of Oncogenic Signaling Molecules and Secreted Growth Factors
title_full Protein Synthesis Inhibition Activity of Mesothelin Targeting Immunotoxin LMB-100 Decreases Concentrations of Oncogenic Signaling Molecules and Secreted Growth Factors
title_fullStr Protein Synthesis Inhibition Activity of Mesothelin Targeting Immunotoxin LMB-100 Decreases Concentrations of Oncogenic Signaling Molecules and Secreted Growth Factors
title_full_unstemmed Protein Synthesis Inhibition Activity of Mesothelin Targeting Immunotoxin LMB-100 Decreases Concentrations of Oncogenic Signaling Molecules and Secreted Growth Factors
title_short Protein Synthesis Inhibition Activity of Mesothelin Targeting Immunotoxin LMB-100 Decreases Concentrations of Oncogenic Signaling Molecules and Secreted Growth Factors
title_sort protein synthesis inhibition activity of mesothelin targeting immunotoxin lmb-100 decreases concentrations of oncogenic signaling molecules and secreted growth factors
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6267581/
https://www.ncbi.nlm.nih.gov/pubmed/30384408
http://dx.doi.org/10.3390/toxins10110447
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