Specific Attenuation of Purinergic Signaling during Bortezomib-Induced Peripheral Neuropathy In Vitro

Holzer, Anna-Katharina; Suciu, Ilinca; Karreman, Christiaan; Goj, Thomas; Leist, Marcel

Specific Attenuation of Purinergic Signaling during Bortezomib-Induced Peripheral Neuropathy In Vitro

Dateien

Holzer_2-1d8cr5x5k5vr32.pdfGröße: 2.3 MBDownloads: 81

Datum

2022

Autor:innen

Holzer, Anna-Katharina

Suciu, Ilinca

Karreman, Christiaan

Goj, Thomas

Leist, Marcel

Publikationstyp

Zeitschriftenartikel

Publikationsstatus

Published

Erschienen in

International Journal of Molecular Sciences. MDPI. 2022, 23(7), 3734. ISSN 1661-6596. eISSN 1422-0067. Available under: doi: 10.3390/ijms23073734

Zusammenfassung

Human peripheral neuropathies are poorly understood, and the availability of experimental models limits further research. The PeriTox test uses immature dorsal root ganglia (DRG)-like neurons, derived from induced pluripotent stem cells (iPSC), to assess cell death and neurite damage. Here, we explored the suitability of matured peripheral neuron cultures for the detection of sub-cytotoxic endpoints, such as altered responses of pain-related P2X receptors. A two-step differentiation protocol, involving the transient expression of ectopic neurogenin-1 (NGN1) allowed for the generation of homogeneous cultures of sensory neurons. After >38 days of differentiation, they showed a robust response (Ca²⁺-signaling) to the P2X3 ligand α,β-methylene ATP. The clinical proteasome inhibitor bortezomib abolished the P2X3 signal at ≥5 nM, while 50–200 nM was required in the PeriTox test to identify neurite damage and cell death. A 24 h treatment with low nM concentrations of bortezomib led to moderate increases in resting cell intracellular Ca²⁺ concentration but signaling through transient receptor potential V1 (TRPV1) receptors or depolarization-triggered Ca²⁺ influx remained unaffected. We interpreted the specific attenuation of purinergic signaling as a functional cell stress response. A reorganization of tubulin to form dense structures around the cell somata confirmed a mild, non-cytotoxic stress triggered by low concentrations of bortezomib. The proteasome inhibitors carfilzomib, delanzomib, epoxomicin, and MG-132 showed similar stress responses. Thus, the model presented here may be used for the profiling of new proteasome inhibitors in regard to their side effect (neuropathy) potential, or for pharmacological studies on the attenuation of their neurotoxicity. P2X3 signaling proved useful as endpoint to assess potential neurotoxicants in peripheral neurons.

Fachgebiet (DDC)

570 Biowissenschaften, Biologie

Schlagwörter

nociceptors; purinergic receptor P2X3; peripheral nervous system diseases; bortezomib; proteasome inhibitors

Zitieren

ISO 690

HOLZER, Anna-Katharina, Ilinca SUCIU, Christiaan KARREMAN, Thomas GOJ, Marcel LEIST, 2022. Specific Attenuation of Purinergic Signaling during Bortezomib-Induced Peripheral Neuropathy In Vitro. In: International Journal of Molecular Sciences. MDPI. 2022, 23(7), 3734. ISSN 1661-6596. eISSN 1422-0067. Available under: doi: 10.3390/ijms23073734

BibTex

@article{Holzer2022-04Speci-57242,
  year={2022},
  doi={10.3390/ijms23073734},
  title={Specific Attenuation of Purinergic Signaling during Bortezomib-Induced Peripheral Neuropathy In Vitro},
  number={7},
  volume={23},
  issn={1661-6596},
  journal={International Journal of Molecular Sciences},
  author={Holzer, Anna-Katharina and Suciu, Ilinca and Karreman, Christiaan and Goj, Thomas and Leist, Marcel},
  note={Article Number: 3734}
}

RDF

<rdf:RDF
    xmlns:dcterms="http://purl.org/dc/terms/"
    xmlns:dc="http://purl.org/dc/elements/1.1/"
    xmlns:rdf="http://www.w3.org/1999/02/22-rdf-syntax-ns#"
    xmlns:bibo="http://purl.org/ontology/bibo/"
    xmlns:dspace="http://digital-repositories.org/ontologies/dspace/0.1.0#"
    xmlns:foaf="http://xmlns.com/foaf/0.1/"
    xmlns:void="http://rdfs.org/ns/void#"
    xmlns:xsd="http://www.w3.org/2001/XMLSchema#" > 
  <rdf:Description rdf:about="https://kops.uni-konstanz.de/server/rdf/resource/123456789/57242">
    <dcterms:abstract xml:lang="eng">Human peripheral neuropathies are poorly understood, and the availability of experimental models limits further research. The PeriTox test uses immature dorsal root ganglia (DRG)-like neurons, derived from induced pluripotent stem cells (iPSC), to assess cell death and neurite damage. Here, we explored the suitability of matured peripheral neuron cultures for the detection of sub-cytotoxic endpoints, such as altered responses of pain-related P2X receptors. A two-step differentiation protocol, involving the transient expression of ectopic neurogenin-1 (NGN1) allowed for the generation of homogeneous cultures of sensory neurons. After &gt;38 days of differentiation, they showed a robust response (Ca&lt;sup&gt;2+&lt;/sup&gt;-signaling) to the P2X3 ligand α,β-methylene ATP. The clinical proteasome inhibitor bortezomib abolished the P2X3 signal at ≥5 nM, while 50–200 nM was required in the PeriTox test to identify neurite damage and cell death. A 24 h treatment with low nM concentrations of bortezomib led to moderate increases in resting cell intracellular Ca&lt;sup&gt;2+&lt;/sup&gt; concentration but signaling through transient receptor potential V1 (TRPV1) receptors or depolarization-triggered Ca&lt;sup&gt;2+&lt;/sup&gt; influx remained unaffected. We interpreted the specific attenuation of purinergic signaling as a functional cell stress response. A reorganization of tubulin to form dense structures around the cell somata confirmed a mild, non-cytotoxic stress triggered by low concentrations of bortezomib. The proteasome inhibitors carfilzomib, delanzomib, epoxomicin, and MG-132 showed similar stress responses. Thus, the model presented here may be used for the profiling of new proteasome inhibitors in regard to their side effect (neuropathy) potential, or for pharmacological studies on the attenuation of their neurotoxicity. P2X3 signaling proved useful as endpoint to assess potential neurotoxicants in peripheral neurons.</dcterms:abstract>
    <bibo:uri rdf:resource="https://kops.uni-konstanz.de/handle/123456789/57242"/>
    <dc:contributor>Holzer, Anna-Katharina</dc:contributor>
    <dc:creator>Leist, Marcel</dc:creator>
    <dc:contributor>Goj, Thomas</dc:contributor>
    <dc:creator>Karreman, Christiaan</dc:creator>
    <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/28"/>
    <dspace:hasBitstream rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/57242/1/Holzer_2-1d8cr5x5k5vr32.pdf"/>
    <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2022-04-08T13:18:11Z</dcterms:available>
    <dc:contributor>Karreman, Christiaan</dc:contributor>
    <dc:contributor>Suciu, Ilinca</dc:contributor>
    <dc:creator>Goj, Thomas</dc:creator>
    <dcterms:issued>2022-04</dcterms:issued>
    <dcterms:rights rdf:resource="http://creativecommons.org/licenses/by/4.0/"/>
    <dc:language>eng</dc:language>
    <foaf:homepage rdf:resource="http://localhost:8080/"/>
    <dcterms:hasPart rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/57242/1/Holzer_2-1d8cr5x5k5vr32.pdf"/>
    <dc:creator>Holzer, Anna-Katharina</dc:creator>
    <dcterms:title>Specific Attenuation of Purinergic Signaling during Bortezomib-Induced Peripheral Neuropathy In Vitro</dcterms:title>
    <dc:contributor>Leist, Marcel</dc:contributor>
    <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2022-04-08T13:18:11Z</dc:date>
    <void:sparqlEndpoint rdf:resource="http://localhost/fuseki/dspace/sparql"/>
    <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/28"/>
    <dc:creator>Suciu, Ilinca</dc:creator>
    <dc:rights>Attribution 4.0 International</dc:rights>
  </rdf:Description>
</rdf:RDF>

Universitätsbibliographie

Ja

Begutachtet

Ja