Domperidone Protects Cells from Intoxication with <i>Clostridioides difficile</i> Toxins by Inhibiting Hsp70-Assisted Membrane Translocation
<i>Clostridioides difficile</i> infections cause severe symptoms ranging from diarrhea to pseudomembranous colitis due to the secretion of AB-toxins, TcdA and TcdB. Both toxins are taken up into cells through receptor-mediated endocytosis, autoproteolytic processing and translocation of...
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MDPI AG
2023-06-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/15/6/384 |
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| author | Maria Braune-Yan Jinfang Jia Mary Wahba Johannes Schmid Panagiotis Papatheodorou Holger Barth Katharina Ernst |
| author_facet | Maria Braune-Yan Jinfang Jia Mary Wahba Johannes Schmid Panagiotis Papatheodorou Holger Barth Katharina Ernst |
| author_sort | Maria Braune-Yan |
| collection | DOAJ |
| description | <i>Clostridioides difficile</i> infections cause severe symptoms ranging from diarrhea to pseudomembranous colitis due to the secretion of AB-toxins, TcdA and TcdB. Both toxins are taken up into cells through receptor-mediated endocytosis, autoproteolytic processing and translocation of their enzyme domains from acidified endosomes into the cytosol. The enzyme domains glucosylate small GTPases such as Rac1, thereby inhibiting processes such as actin cytoskeleton regulation. Here, we demonstrate that specific pharmacological inhibition of Hsp70 activity protected cells from TcdB intoxication. In particular, the established inhibitor VER-155008 and the antiemetic drug domperidone, which was found to be an Hsp70 inhibitor, reduced the number of cells with TcdB-induced intoxication morphology in HeLa, Vero and intestinal CaCo-2 cells. These drugs also decreased the intracellular glucosylation of Rac1 by TcdB. Domperidone did not inhibit TcdB binding to cells or enzymatic activity but did prevent membrane translocation of TcdB’s glucosyltransferase domain into the cytosol. Domperidone also protected cells from intoxication with TcdA as well as CDT toxin produced by hypervirulent strains of <i>Clostridioides difficile</i>. Our results reveal Hsp70 requirement as a new aspect of the cellular uptake mechanism of TcdB and identified Hsp70 as a novel drug target for potential therapeutic strategies required to combat severe <i>Clostridioides difficile</i> infections. |
| first_indexed | 2024-03-11T01:52:24Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-11T01:52:24Z |
| publishDate | 2023-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-b95382d66fb94765915a82c4241d810c2023-11-18T12:55:38ZengMDPI AGToxins2072-66512023-06-0115638410.3390/toxins15060384Domperidone Protects Cells from Intoxication with <i>Clostridioides difficile</i> Toxins by Inhibiting Hsp70-Assisted Membrane TranslocationMaria Braune-Yan0Jinfang Jia1Mary Wahba2Johannes Schmid3Panagiotis Papatheodorou4Holger Barth5Katharina Ernst6Institute of Experimental and Clinical Pharmacology, Toxicology and Pharmacology of Natural Products, Ulm University Medical Center, 89081 Ulm, GermanyInstitute of Experimental and Clinical Pharmacology, Toxicology and Pharmacology of Natural Products, Ulm University Medical Center, 89081 Ulm, GermanyInstitute of Experimental and Clinical Pharmacology, Toxicology and Pharmacology of Natural Products, Ulm University Medical Center, 89081 Ulm, GermanyInstitute of Experimental and Clinical Pharmacology, Toxicology and Pharmacology of Natural Products, Ulm University Medical Center, 89081 Ulm, GermanyInstitute of Experimental and Clinical Pharmacology, Toxicology and Pharmacology of Natural Products, Ulm University Medical Center, 89081 Ulm, GermanyInstitute of Experimental and Clinical Pharmacology, Toxicology and Pharmacology of Natural Products, Ulm University Medical Center, 89081 Ulm, GermanyInstitute of Experimental and Clinical Pharmacology, Toxicology and Pharmacology of Natural Products, Ulm University Medical Center, 89081 Ulm, Germany<i>Clostridioides difficile</i> infections cause severe symptoms ranging from diarrhea to pseudomembranous colitis due to the secretion of AB-toxins, TcdA and TcdB. Both toxins are taken up into cells through receptor-mediated endocytosis, autoproteolytic processing and translocation of their enzyme domains from acidified endosomes into the cytosol. The enzyme domains glucosylate small GTPases such as Rac1, thereby inhibiting processes such as actin cytoskeleton regulation. Here, we demonstrate that specific pharmacological inhibition of Hsp70 activity protected cells from TcdB intoxication. In particular, the established inhibitor VER-155008 and the antiemetic drug domperidone, which was found to be an Hsp70 inhibitor, reduced the number of cells with TcdB-induced intoxication morphology in HeLa, Vero and intestinal CaCo-2 cells. These drugs also decreased the intracellular glucosylation of Rac1 by TcdB. Domperidone did not inhibit TcdB binding to cells or enzymatic activity but did prevent membrane translocation of TcdB’s glucosyltransferase domain into the cytosol. Domperidone also protected cells from intoxication with TcdA as well as CDT toxin produced by hypervirulent strains of <i>Clostridioides difficile</i>. Our results reveal Hsp70 requirement as a new aspect of the cellular uptake mechanism of TcdB and identified Hsp70 as a novel drug target for potential therapeutic strategies required to combat severe <i>Clostridioides difficile</i> infections.https://www.mdpi.com/2072-6651/15/6/384toxin uptakechaperonesHsp70domperidone<i>Clostridioides difficile</i> toxinstoxin inhibitors |
| spellingShingle | Maria Braune-Yan Jinfang Jia Mary Wahba Johannes Schmid Panagiotis Papatheodorou Holger Barth Katharina Ernst Domperidone Protects Cells from Intoxication with <i>Clostridioides difficile</i> Toxins by Inhibiting Hsp70-Assisted Membrane Translocation Toxins toxin uptake chaperones Hsp70 domperidone <i>Clostridioides difficile</i> toxins toxin inhibitors |
| title | Domperidone Protects Cells from Intoxication with <i>Clostridioides difficile</i> Toxins by Inhibiting Hsp70-Assisted Membrane Translocation |
| title_full | Domperidone Protects Cells from Intoxication with <i>Clostridioides difficile</i> Toxins by Inhibiting Hsp70-Assisted Membrane Translocation |
| title_fullStr | Domperidone Protects Cells from Intoxication with <i>Clostridioides difficile</i> Toxins by Inhibiting Hsp70-Assisted Membrane Translocation |
| title_full_unstemmed | Domperidone Protects Cells from Intoxication with <i>Clostridioides difficile</i> Toxins by Inhibiting Hsp70-Assisted Membrane Translocation |
| title_short | Domperidone Protects Cells from Intoxication with <i>Clostridioides difficile</i> Toxins by Inhibiting Hsp70-Assisted Membrane Translocation |
| title_sort | domperidone protects cells from intoxication with i clostridioides difficile i toxins by inhibiting hsp70 assisted membrane translocation |
| topic | toxin uptake chaperones Hsp70 domperidone <i>Clostridioides difficile</i> toxins toxin inhibitors |
| url | https://www.mdpi.com/2072-6651/15/6/384 |
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