Background & Aims

Oxaliplatin (OXL) is a third-generation platinum-based antineoplastic agent, utilized as the first-line treatment for colorectal cancer (CRC; the 3rd most common cancer), of paramount importance in oncology clinics. Its primary side effect is peripheral sensory neuropathy (PSN), a debilitating and painful neurotoxic condition that may result in treatment cessation or dose reduction, consequently diminishing the survival rates of CRC patients (PATCHMAN et al., 2015; TENG et al., 2022; GODINHO et al., 2020). To date there are no efficient therapy to manage OXL PSN (CAVALETTI et al., 2020). McLTP1, a protein isolated from Morinda citrifolia seeds, from previous studies, has been shown to have an antinociceptive, anti-inflammatory and antioxidant effect in preclinical studies by oral route (CAMPOS et al., 2016, 2017; DO CARMO, et al., 2022). Therefore, this study sought to investigate the effect of McLTP1 on oxaliplatin-induced PSN in mice and further in CT26 tumor bearing mice.

Methods

PSN was induced by OXL injections (2 mg/kg, i.v., 2x/weeks, in total 9 injections) for 28 days in Swiss mice (male, 25-30g). The effect of McLTP1 (1 – 4 mg/kg) was evaluated, up to the 28th day, through nociceptive tests: Von Frey, cold thermal allodynia, and other behavioral tests (Rota rod and Open field test). Involvement of TRPA1 or TRPM8 was evaluated, as well as the morphologic changes in sciatic nerve, c-Fos and ATF-3 positive area in DRG and spinal cord and oxidative stress markers (nitrotyrosine, MDA and GSH). Furthermore, McLTP1 effect was analyzed in CT26 (Mice colorectal cancer cell line) tumor bearing mice to investigate the possible interference in OXL antitumor effect. For this CT26 cells were cultured and injected subcutaneously (1×10ˆ6) in the right flank of BALB/c mice. When the tumor mass reached 50–100mm3, animals were injected with OXL at every 3 days. McLTP1 was injected diary. Tumor was measured every 3 days until 21th day and collected for histopathology.

Results

OXL reduced (p<0.05) the paw withdrawal threshold (von frey) and increased cold allodinia up to 39.6 and 77.%, respectively. McLTP1 (4 mg/kg) prevented (p < 0.05) these nociceptive responses up to 20.5 and 74.9%. TRPA1 receptor agonist did not interfere with McLTP1 inhibitory effect, while with TRPM8 receptor agonist reversed this effect by 65.1% (p<0.05). OXL reduced (p<0.05) GSH levels in the sciatic nerve (SN) and spinal cord (SC) by 82.5% and 27.8%, respectively, and McLTP1, prevented (p<0.05) this decreases by 37.8% and 98.9%. OXL increased (p<0.05) MDA levels in the SN and SC by 60.97% and 34.27%, respectively, and McLTP1, prevented (p<0.05) this increase by 36.33% and 42.3%. OXL reduced (p<0.05) total leucocyte number (57.96%) and McLTP1 prevented (p<0.05) this decrease 47%, mainly the lymphocytes (42.2%). OXL increased (p<0.05) the c-Fos expression in dorsal root ganglia (DRG; 82%) and SC (75%) and increased ATF3 expression in DRG (93%). McLTP decreased (p<0.05) both c-FOS and ATF3 in DRG by 80.1 and 95%, respectively and c-FOS in SC by 79.2%. OXL also increased (p<0.05) by 63% nitrotyrosine expression in DRG and McLPT1 decreased this effect by 46%. In CT26 tumor bearing mice OXL decreased tumor volume by 34% and McLPT1 did not interfere with this effect.

Conclusions

In conclusion, the data suggest that daily administration of McLTP1 (4 mg/kg) in mice subjected to peripheral neuropathy due to OXL: (1) has an antinociceptive effect which shows the possible participation of TRPM8 receptors, but not of TRPA1; (2) has a neuroprotective and modulatory action upon oxidative stress on DRG, SN and SC; (3) prevents lymphopenia caused by oxaliplatin. Additionally McLTP1 does not interfere with OXL antitumoral effect in colorectal CT26 tumor bearing mice, showing a promissor alternative in prevention of OXL associated peripheral sensitive neuropathy.

References

CAMPOS, D. C. O. et al. First isolation and antinociceptive activity of a lipid transfer proteinfrom noni (Morinda citrifolia) seeds. Internat. J. Biolog. Macromol. v. 86, p. 71–79, 2016.
CAMPOS, D. C. O. et al. Morinda citrifolia lipid transfer protein 1 exhibits anti-inflammatory activity by modulation of pro- and anti-inflammatory cytokines. Int J Biol Macromol, v. 103, p. 1121-1129, 2017.
CAVALETTI, G., MARMIROLI, P. Management of Oxaliplatin-Induced Peripheral Sensory Neuropathy. Cancers (Basel), v. 12, n. 6, p. 1370, 2020.
DO CARMO, L. D. et al. Therapeutic effects of a lipid transfer protein isolated from Morinda citrifolia L. (noni) seeds on irinotecan-induced intestinal mucositis in mice. Naunyn Schmiedebergs Arch Pharmacol., v. 395, n. 9, p. 1097-1107, 2022.
GODINHO, P. A. R. et al. FLOX (5-fluorouracil?+?leucovorin?+?oxaliplatin) chemotherapy for colorectal cancer leads to long-term orofacial neurotoxicity: a STROBE-guided longitudinal prospective study. Int J Clin Oncol., v. 25, n. 12, p. 2066-2074, 2020
PACHMAN D.R. et al. Clinical course of oxaliplatin-induced neuropathy: results from the randomized phase III trial N08CB (Alliance). J Clin Oncol 33(30):3416–3422, 2015.
TENG C, et al. Systematic review of long-term chemotherapy-induced peripheral neuropathy (CIPN) following adjuvant oxaliplatin for colorectal cancer. Support Care Cancer. Jan;30(1):33-47, 2022.

Presenting Author

Mariana Vale

Poster Authors

Mariana Vale

PhD

Federal University of Ceará

Lead Author

E-mail

Francisco Rafael Alves Santana Cesario

PhD

Federal University of Ceará

Lead Author

E-mail

Breno Leonardo Da Silva Mansur Abucater

Bachelor's Degree

Federal University of Ceará

Lead Author

E-mail

Jonas Costa de França

Bachelor's Degree

Federal University of Ceará

Lead Author

E-mail

Anamaria Falcão Pereira de Souza

PhD

Federal University of Ceará

Lead Author

E-mail

Cristiane Maria Pereira da Silva

Master's degree

Federal University of Ceará

Lead Author

E-mail

Hermógenes David de Oliveira

PhD

Federal University of Ceará

Lead Author

E-mail

Ellen Dayane Dantas Rodrigues

Undergraduate

Lead Author

Paula Tais Gomes Muniz

Undergraduate

Lead Author

Bianca de Souza Bezerra

Undergraduate

Lead Author

Victor Machado de Carvalho

Master's Student

Lead Author

Topics

  • Specific Pain Conditions/Pain in Specific Populations: Cancer Pain & Palliative Care