Genetic and cellular sensitivity of Caenorhabditis elegans to the chemotherapeutic agent cisplatin

Abstract

Cisplatin and derivatives are commonly used as chemotherapeutic agents. Although the cytotoxic action of cisplatin on cancer cells is very efficient, clinical oncologists need to deal with two major difficulties, namely the onset of resistance to the drug and the cytotoxic effect in patients. Here, we used Caenorhabditis elegans to investigate factors influencing the response to cisplatin in multicellular organisms. In this hermaphroditic model organism, we observed that sperm failure is a major cause of cisplatin-induced infertility. RNA sequencing data indicate that cisplatin triggers a systemic stress response, in which DAF-16/FOXO and SKN-1/NRF2, two conserved transcription factors, are key regulators. We determined that inhibition of the DNA damage-induced apoptotic pathway does not confer cisplatin protection to the animal. However, mutants for the pro-apoptotic BH3-only gene ced-13 are sensitive to cisplatin, suggesting a protective role of the intrinsic apoptotic pathway. Finally, we demonstrated that our system can also be used to identify mutations providing resistance to cisplatin and therefore potential biomarkers of innate cisplatin-refractory patients. We show that mutants for the redox regulator trxr-1, ortholog of the mammalian thioredoxin reductase 1 TRXR1, display cisplatin resistance. By CRISPR/Cas9, we determined that such resistance relies on the presence of the single selenocysteine residue in TRXR-1.

Alternative redox forms of ASNA-1 separate insulin signaling from tail-anchored protein targeting and cisplatin resistance in C. elegans

“…Its mammalian homolog can be targeted by cisplatin to form cytotoxic SecTRAPS 59 . Work in C. elegans shows that TRXR-1 confers systemic sensitivity to cisplatin via the selenocysteine residue in larval stage animals displaying mitotic proliferation 60 .…”

The glutathione system and the related thiol network in Caenorhabditis elegans

“…Having only one selenoprotein has made C. elegans a valuable in vivo model to investigate dose-dependent beneficial and toxicological properties of selenium-based compounds [ 63 , [164] , [165] , [166] ]. The CRISPR/Cas9 system has been used to generate targeted mutations of the trxr-1 selenocysteine [ 167 ]. Two such trxr-1 variants; one with a point mutation where the selenocysteine residue was replaced by a cysteine and the other with a premature stop codon, showed resistance during development when exposed to the chemotherapeutic agent, cisplatin [ 167 ].…”

“…The CRISPR/Cas9 system has been used to generate targeted mutations of the trxr-1 selenocysteine [ 167 ]. Two such trxr-1 variants; one with a point mutation where the selenocysteine residue was replaced by a cysteine and the other with a premature stop codon, showed resistance during development when exposed to the chemotherapeutic agent, cisplatin [ 167 ]. The trxr-1 gene affords partial protection from degeneration of dopaminergic neurons against the neurotoxin 6-hydroxydopamine (6-OHDA) [ 142 ].…”

Insights into cisplatin-induced neurotoxicity and mitochondrial dysfunction in Caenorhabditis elegans

“…The C. elegans BH3-only protein CED-13 promotes cell survival in response to mtROS, instead of inducing apoptosis (Yee et al, 2014). Moreover, as we have previously described, CED-13 protects C. elegans from cisplatin-induced toxicity (García-Rodríguez et al, 2018) (Fig. 2B).…”

“…This nematode is a well-established system to investigate neuromodulatory pathways in vivo (Van Damme et al, 2021) and the effect of genotoxic drugs in a pluricellular context (Honnen, 2017; Kaletta and Hengartner, 2006). Particularly, we and others have recently demonstrated the value of C. elegans to explore genetic, cellular, and molecular factors involved in cisplatin response (Hemmingsson et al, 2010; García-Rodríguez et al, 2018; Wellenberg et al, 2021). Here, exploiting our previous methodologies, along with CRISPR-Cas9 technology and semi-automated platforms (Seahorse XFe96 Analyzer and Tierpsy Tracker), we further investigated the effects of cisplatin-based therapies.…”