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. 2023 Sep 1;164(9):2060-2069.
doi: 10.1097/j.pain.0000000000002906. Epub 2023 Apr 19.

TRPM3 as a novel target to alleviate acute oxaliplatin-induced peripheral neuropathic pain

Vincenzo Davide Aloi  1   2   3 , Sílvia João Poseiro Coutinho Pinto  2   3 , Rita Van Bree  1 , Katrien Luyten  1 , Thomas Voets  2   3 , Joris Vriens  1   2
Affiliations

TRPM3 as a novel target to alleviate acute oxaliplatin-induced peripheral neuropathic pain

Vincenzo Davide Aloi et al. Pain. .

Abstract

Chemotherapy-induced peripheral neuropathic pain (CIPNP) is an adverse effect observed in up to 80% of patients of cancer on treatment with cytostatic drugs including paclitaxel and oxaliplatin. Chemotherapy-induced peripheral neuropathic pain can be so severe that it limits dose and choice of chemotherapy and has significant negative consequences on the quality of life of survivors. Current treatment options for CIPNP are limited and unsatisfactory. TRPM3 is a calcium-permeable ion channel functionally expressed in peripheral sensory neurons involved in the detection of thermal stimuli. Here, we focus on the possible involvement of TRPM3 in acute oxaliplatin-induced mechanical allodynia and cold hypersensitivity. In vitro calcium microfluorimetry and whole-cell patch-clamp experiments showed that TRPM3 is functionally upregulated in both heterologous and homologous expression systems after acute (24 hours) oxaliplatin treatment, whereas the direct application of oxaliplatin was without effect. In vivo behavioral studies using an acute oxaliplatin model for CIPNP showed the development of cold and mechano hypersensitivity in control mice, which was lacking in TRPM3 deficient mice. In addition, the levels of protein ERK, a marker for neuronal activity, were significantly reduced in dorsal root ganglion neurons derived from TRPM3 deficient mice compared with control after oxaliplatin administration. Moreover, intraperitoneal injection of a TRPM3 antagonist, isosakuranetin, effectively reduced the oxaliplatin-induced pain behavior in response to cold and mechanical stimulation in mice with an acute form of oxaliplatin-induced peripheral neuropathy. In summary, TRPM3 represents a potential new target for the treatment of neuropathic pain in patients undergoing chemotherapy.

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Conflict of interest statement

The authors have no conflicts of interest to declare.

Sponsorships or competing interests that may be relevant to content are disclosed at the end of this article.

Figures

Figure 1.
Figure 1.
Modulation of TRPM3 function in HEK293T cells stably expressing murine TRPM3 after oxaliplatin pretreatment. (A) Time course of intracellular calcium concentrations ([Ca2+]i) (mean ± SEM) at 37°C on application of the TRPM3 inhibitors primidone (100 µM) in HEK-mTRPM3 cells after pretreatment with oxaliplatin (100 µM) (n = 299) and vehicle (n = 293) and nontransfected (NT) cells (n = 97) (N = 3 independent experiments). (B) Basal intracellular calcium concentrations before (full bars) and after the application of primidone (open bars). Data are represented as mean ± SEM. Statistically significant changes in the basal channel activity were assessed using a Kruskal–Wallis ANOVA with the Dunn post hoc test, where *P = <0.05 and ***P = <0.001. (C and E) Time course of intracellular calcium concentrations ([Ca2+]i) (mean ± SEM) for HEK-mTRPM3 cells in response to PS (40 µM) and heat (37°C) after oxaliplatin (100 µM) and vehicle treatment. (D and F) Quantification of calcium responses for experiments as in panel (C and E), respectively. Where ***P < 0.001 (Mann–Whitney U test). ANOVA, analysis of variance; NT, nontransfected; PS, pregnenolone sulfate; Vhc, vehicle; OXA, oxaliplatin.
Figure 2.
Figure 2.
Modulation of TRPM3 function in HEK293-mTRPM3. (A) Time course at ± 150 mV of a whole-cell patch-clamp recording showing the effect of pregnenolone sulphate (PS; 40 µM) on HEK-mTRPM3 expressing cells after vehicle preincubation (n = 16). (B) Current (I)−voltage (V) relationship corresponding to the time points indicated in A. (C) Time course at ± 150 mV of a whole-cell patch-clamp recording showing the effect of PS (40 µM) on HEK-mTRPM3 expressing cells after 24 hours of oxaliplatin (100 µM) preincubation (n = 16). (D) I-V relationship corresponding to the time points indicated in C. (E) Mean current increase at ± 150 mV in HEK-TRPM3 cells on PS application after 24 hours of vehicle or oxaliplatin preincubation. Where *P < 0.05 (Mann–Whitney U test). PS, pregnenolone sulfate.
Figure 3.
Figure 3.
Modulation of TRPM3 function in DRG sensory neurons after oxaliplatin pretreatment. Time course of the intracellular Ca2+ concentration ([Ca2+]i) (mean ± SEM) in DRG isolated from wild-type animals TRPM3+/+ in response to PS (40 µM) and high K+ (50 mM) after oxaliplatin (100 µM; red) and vehicle treatment in black (A) and after 24 hours after oxaliplatin (6 mg/kg; red color) or vehicle i.p injection (black color) (E). (B, C, F, G) Quantification of calcium responses for experiments as in panel A and E, respectively, *** P < 0.001 (Mann–Whitney U test). Percentage of sensory neurons derived from TRPM3+/+ responding stimulation by PS (40 µM) after oxaliplatin and vehicle preincubation (D) and after 24 hours after oxaliplatin (6 mg/kg) or vehicle i.p injection (H). Where *P < 0.05 (χ2 test); **P < 0.01 (χ2 test). DRG, dorsal root ganglia; PS, pregnenolone sulfate.
Figure 4.
Figure 4.
Nocifensive responses to PS after oxaliplatin and vehicle injection. (A) Total duration of nocifensive behavior (paw licks and lifts during a period of 10 minutes) in response to intraplantar injection of pregnenolone sulfate (PS, 5 nmol/paw) in TRPM3+/+ and TRPM3−/− mice (n = 8 animals/genotype). Data are represented as mean ± SEM. Statistically significant changes in the duration of PS-evoked pain responses were assessed using the 2-way ANOVA repeated measurement statistical test with Sidak-holm post hoc test. Where **P < 0.01; ***P < 0.001. ANOVA, analysis of variance; WT, TRPM3+/+; KO, TRPM3−/−.
Figure 5.
Figure 5.
TRPM3 genetic ablation reduces oxaliplatin-induced mechanical allodynia. (A and B) A single intraperitoneal injection of oxaliplatin (6 mg/kg) induces in TRPM3+/+ mice (black color) a time-dependent reduction in mechanical nociceptive threshold and increase in cold sensitivity, respectively. Mechanosensitivity (A) and cold sensitivity (B) were measured at baseline (Day −1), 24 hours after the injection of vehicle (Day 0) and after 6 days (Day 6). On day 7 (Day 7), animals treated with oxaliplatin and mechano (A) or cold sensitivity (B) were measured after 24 hours (Day 8) and after 6 days (Day 13) after injection. The development of mechanical and cold allodynia observed in TRPM3+/+ animals (black) after oxaliplatin treatment was decreased in TRPM3−/− mice (red color). Data are presented as mean ± SEM (n = 8 mice per group). Statistically significant changes were assessed by using the 2-way ANOVA repeated measurement statistical test with the Sidak-Holm post hoc test. Where *P < 0.05; **P < 0.01; ***P < 0.001. ANOVA, analysis of variance; Vhc, vehicle; OXA, oxaliplatin; WT, wild-type; KO, TRPM3−/−.
Figure 6.
Figure 6.
pERK expression levels in DRG of wild-type and TRPM3−/− mice after dosing of oxaliplatin. (A) Sections of pERK immunostaining in DRG neurons of TRPM3-WT and TRPM3-KO mice at 24 hours after oxaliplatin or vehicle injection. Scale bars, 50 μm. (B) The percentage of pERK-positive cells in the DRG sections was counted (n = 6 sections, from 3 independent DRG preparations). Data are expressed as the mean ± SEM. Statistically significant changes were assessed by using the 2-way ANOVA statistical test with the Sidak-Holm post hoc test. Where ***P <0.001. ANOVA, analysis of variance; DRG, dorsal root ganglia; pERK, protein ERK; WT, wild-type; KO, TRPM3−/−.
Figure 7.
Figure 7.
Pharmacological inhibition of TRPM3 reduces neuropathic pain induced by oxaliplatin. The effect is tested in the electronic Von Frey assay and acetone spray test at day 6 after oxaliplatin treatment. (A and B) Effect of isosakuranetin (2 mg/kg) and tramadol (5 mg/kg) as positive control and vehicle used in the intervention protocol on nociceptive mechanical threshold and escape behavior score in mice treated with oxaliplatin. Results are shown as mean ± SEM of n = 8 mice per condition. Statistical analysis: 2-way ANOVA repeated measurements with the Sidak-holm post hoc test. Where §§ Vhc vs tramadol (P <0.01); ## Vhc vs isosakuranetin (P <0.01); # Vhc vs isosakuranetin (P <0.05). ANOVA, analysis of variance.
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References

    1. Banach M, Juranek JK, Zygulska AL. Chemotherapy-induced neuropathies-a growing problem for patients and health care providers. Brain Behav 2017;7:e00558. - PMC - PubMed
    1. Brzezinski K. Chemotherapy-induced polyneuropathy. Part I. Pathophysiology. Contemp Oncol (Pozn) 2012;16:72–8. - PMC - PubMed
    1. Chukyo A, Chiba T, Kambe T, Yamamoto K, Kawakami K, Taguchi K, Abe K. Oxaliplatin-induced changes in expression of transient receptor potential channels in the dorsal root ganglion as a neuropathic mechanism for cold hypersensitivity. Neuropeptides 2018;67:95–101. - PubMed
    1. Descoeur J, Pereira V, Pizzoccaro A, Francois A, Ling B, Maffre V, Couette B, Busserolles J, Courteix C, Noel J, Lazdunski M, Eschalier A, Authier N, Bourinet E. Oxaliplatin-induced cold hypersensitivity is due to remodelling of ion channel expression in nociceptors. EMBO Mol Med 2011;3:266–78. - PMC - PubMed
    1. Fukuda Y, Li Y, Segal RA. A mechanistic understanding of axon degeneration in chemotherapy-induced peripheral neuropathy. Front Neurosci 2017;11:481. - PMC - PubMed

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