Modeling chemotherapy induced neurotoxicity with human induced pluripotent stem cell (iPSC) -derived sensory neurons

Schinke, Christian; Vallone, Valeria Fernández; Ivanov, Andranik; Peng, Yangfan; Körtvelyessy, Péter; Nolte, Luca; Huehnchen, Petra; Beule, Dieter; Stachelscheid, Harald; Boehmerle, Wolfgang; Endres, Matthias

doi:10.1016/j.nbd.2021.105391

Cited by 41 publications

(39 citation statements)

References 79 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In vitro test methods offer many advantages for the study of specific mechanistic and pharmacological aspects of peripheral neurotoxicity due to their relative "simplicity", and as environmental factors can be very tightly controlled. Indeed, iPSC-based in vitro test methods have been repeatedly used to measure the effects of chemotherapeutics on cellular viability or morphology [3,4,27,[70][71][72]. However, there is still a dearth of studies that employ human iPSCderived nociceptor cultures to assess alterations of signaling endpoints.…”

Section: Discussionmentioning

confidence: 99%

Generation of human nociceptor-enriched sensory neurons for the study of pain-related dysfunctions

Holzer

Karreman

Suciu

et al. 2022

Preprint

View full text Add to dashboard Cite

In vitro models of the peripheral nervous system would benefit from further refinements to better support studies on neuropathies. In particular, the assessment of pain-related signals is still difficult in human cell cultures. Here, we harnessed induced pluripotent stem cells (iPSCs) to generate peripheral sensory neurons enriched in nociceptors. The objective was to generate a culture system with signaling endpoints suitable for pharmacological and toxicological studies. Neurons generated by conventional differentiation protocols expressed moderate levels of P2X3 purinergic receptors and only low levels of TRPV1 capsaicin receptors, when maturation time was kept to the upper practically-useful limit of 6 weeks. As alternative approach, we generated cells with an inducible NGN1 transgene. Ectopic expression of this transcription factor during a defined time window of differentiation resulted in highly-enriched nociceptor cultures, as determined by functional (P2X3 and TRPV1 receptors) and immunocytochemical phenotyping, complemented by extensive transcriptome profiling. Single cell recordings of Ca2+-indicator fluorescence from >9,000 cells were used to establish the "fraction of reactive cells" in a stimulated population as experimental endpoint, that appeared robust, transparent and quantifiable. To provide an example of application to biomedical studies, functional consequences of prolonged exposure to the chemotherapeutic drug oxaliplatin were examined at non-cytotoxic concentrations. We found (i) neuronal (allodynia-like) hypersensitivity to otherwise non-activating mechanical stimulation that could be blocked by modulators of voltage-gated sodium channels; (ii) hyper-responsiveness to TRPV1 receptor stimulation. These findings and several other measured functional alterations indicate that the model is suitable for pharmacological and toxicological studies related to peripheral neuropathies.

show abstract

Section: Discussionmentioning

confidence: 99%

Generation of human nociceptor-enriched sensory neurons for the study of pain-related dysfunctions

Holzer

Karreman

Suciu

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“… 46 As a third example, highly differentiated sensory neurons over a 40-day period were then tested with a series of chemotherapeutic agents for toxicological evaluations. 47 Only in a limited number of cases were studies undertaken that assessed the capacity for cell proliferation and differentiation as seen with MT for IPSCs and NSCs, respectively. There was also a more apparent effort made to discern underlying molecular mechanism pathways in the collective individual stem cell-hormesis areas where such efforts have been substantial.…”

Section: Discussionmentioning

confidence: 99%

“…Of interest, in this regard have been efforts to use IPSC-derived sensory neurons as a disease model of chemotherapy-induced neurotoxicity. Schinke et al 47 have addressed this issue with a focus on whether neurotoxin chemotherapy in clinically relevant steady-state concentrations affects cell viability of IPSC-derived sensory neurons. In their experimental protocol, 40 days were required for neurogenic differentiation to achieve morphologically pure neuronal cultures with ganglia and extensive axons.…”

Section: Assessment Of Hormesis In Induced Pluripotent Stem Cells (Ip...mentioning

confidence: 99%

See 1 more Smart Citation

Induced Pluripotent Stem Cells and Hormesis

Calabrese

2022

Dose-Response

View full text Add to dashboard Cite

This paper represents the first assessment of agent-induced hormetic dose responses in induced pluripotent stem cells and their derived cells. The hormetic dose responses were induced by a broad range of chemicals, including pharmaceuticals (eg, metformin), dietary supplements/extracts from medicinal plants (eg, curcumin), and endogenous agents (eg, melatonin). The paper assesses the mechanistic foundations of these induced hormetic dose responses, their therapeutic implications and comparison with hormetic responses in multiple adult and embryonic stem cells.

show abstract

“…Some test methods are based on human peripheral neurons derived from induced pluripotent stem cells (iPSCs). They have been mostly used to investigate drug effects on neurite morphology or cell viability [28][29][30][31]. Such endpoints correlate with events during full-blown BIPN, such as loss of intra-epidermal nerve fibers and alterations in cytoskeletal structure and impairment of axonal transport [3][4][5][6][7].…”

Section: Introductionmentioning

confidence: 99%

Specific attenuation of purinergic signaling during bortezomib-induced peripheral neuropathy

Holzer

Suciu

Karreman

et al. 2022

Preprint

View full text Add to dashboard Cite

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 detection of sub-cytotoxic endpoints, such as altered responses of pain-related P2X receptors. A 2-step differentiation protocol, involving 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+-signalling) to the P2X3 ligand α,β-methylene ATP. The clinical proteasome inhibitor bortezomib abolished the P2X3 signal at ≥ 5 nM, while 50-200 nM were 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+], but signalling through transient receptor potential-V1 (TRPV1) receptors or depolarization-triggered Ca2+-influx remained unaffected. We interpret the specific attenuation of purinergic signalling as functional cell stress response. A reorganization of tubulin to dense structures around the cell somata confirmed a mild, non-cytotoxic stress triggered by low concentrations of bortezomib. The proteasome inhibitors carfilzomib, delanzomib, epoxomycin and MG-132 showed similar stress responses. Thus, the model presented here may be used for profiling of new proteasome inhibitors as to their side effect (neuropathy) potential, or for pharmacological studies on the attenuation of their neurotoxicity. P2X3 signalling proved useful as endpoint to assess potential neurotoxicants in peripheral neurons.

show abstract

Modeling chemotherapy induced neurotoxicity with human induced pluripotent stem cell (iPSC) -derived sensory neurons

Cited by 41 publications

References 79 publications

Generation of human nociceptor-enriched sensory neurons for the study of pain-related dysfunctions

Generation of human nociceptor-enriched sensory neurons for the study of pain-related dysfunctions

Induced Pluripotent Stem Cells and Hormesis

Specific attenuation of purinergic signaling during bortezomib-induced peripheral neuropathy

Contact Info

Product

Resources

About