Targeting RAC1 reactivates pyroptosis to reverse paclitaxel resistance in ovarian cancer by suppressing P21-activated kinase 4

Abstract

Pyroptosis may play an important role in the resistance of ovarian cancer (OC) to chemotherapy. However, the mechanism by which pyroptosis modulation can attenuate chemotherapy resistance has not been comprehensively studied in OC. Here, we demonstrated that RAS-associated C3 botulinum toxin substrate 1 (RAC1) is highly expressed in OC and is negatively correlated with patient outcomes. Through cell function tests and in vivo tumor formation tests, we found that RAC1 can promote tumor growth by mediating paclitaxel (PTX) resistance. RAC1 can mediate OC progression by inhibiting pyroptosis, as evidenced by high-throughput automated confocal imaging, the release of lactate dehydrogenase (LDH), the expression of the inflammatory cytokines IL-1β/IL-18 and the nucleotide oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome. Mechanically, RNA-seq, gene set enrichment analysis (GSEA), coimmunoprecipitation (Co-IP), mass spectrometry (MS), and ubiquitination tests further confirmed that RAC1 inhibits caspase-1/gasdermin D (GSDMD)-mediated canonical pyroptosis through the P21-activated kinase 4 (PAK4)/mitogen-activated protein kinase (MAPK) pathway, thereby promoting PTX resistance in OC cells. Finally, the whole molecular pathway was verified by the results of in vivo drug combination tests, clinical specimen detection and the prognosis. In summary, our results suggest that the combination of RAC1 inhibitors with PTX can reverse PTX resistance by inducing pyroptosis through the PAK4/MAPK pathway.

Keywords

ovarian cancer / paclitaxel resistance / pyroptosis / RAC1

References

[1] Matulonis UA, Sood AK, Fallowfield L, Howitt BE, Sehouli J, Karlan BY. Ovarian cancer. Nat Rev Dis Primers. 2016; 2: 16061.

[2] Kandalaft LE, Dangaj Laniti D, Coukos G. Immunobiology of high-grade serous ovarian cancer: lessons for clinical translation. Nat Rev Cancer. 2022; 22(11): 640-656.

[3] Eisenhauer EA. Real-world evidence in the treatment of ovarian cancer. Ann Oncol. 2017; 28(suppl_8): viii61-viii65.

[4] Richardson DL, Eskander RN, O’Malley DM. Advances in ovarian cancer care and unmet treatment needs for patients with platinum resistance: a narrative review. JAMA Oncol. 2023; 9(6): 851-859.

[5] O’Malley DM. New therapies for ovarian cancer. J Natl Compr Canc Netw. 2019; 17(5.5): 619-621.

[6] Parmar MK, Ledermann JA, Colombo N, et al. Paclitaxel plus platinum-based chemotherapy versus conventional platinum-based chemotherapy in women with relapsed ovarian cancer: the ICON4/AGO-OVAR-2.2 trial. Lancet. 2003; 361(9375): 2099-2106.

[7] Vermorken JB. The integration of paclitaxel and new platinum compounds in the treatment of advanced ovarian cancer. Int J Gynecol Cancer. 2001; 11(suppl 1): 21-30.

[8] Das T, Anand U, Pandey SK, et al. Therapeutic strategies to overcome taxane resistance in cancer. Drug Resist Updat. 2021; 55: 100754.

[9] Lee JP, Hahn HS, Hwang SJ, et al. Selective cyclooxygenase inhibitors increase paclitaxel sensitivity in taxane-resistant ovarian cancer by suppressing P-glycoprotein expression. J Gynecol Oncol. 2013; 24(3): 273-279.

[10] Zheng Z, Liu B, Wu X. RhoGDI2 up-regulates P-glycoprotein expression via Rac1 in gastric cancer cells. Cancer Cell Int. 2015; 15: 41.

[11] Cardama GA, Alonso DF, Gonzalez N, et al. Relevance of small GTPase Rac1 pathway in drug and radio-resistance mechanisms: opportunities in cancer therapeutics. Crit Rev Oncol Hematol. 2018; 124: 29-36.

[12] Yuan Y, Zhang H, Li D, et al. PAK4 in cancer development: emerging player and therapeutic opportunities. Cancer Lett. 2022; 545: 215813.

[13] Santiago-Gómez A, Kedward T, Simões BM, et al. PAK4 regulates stemness and progression in endocrine resistant ER-positive metastatic breast cancer. Cancer Lett. 2019; 458: 66-75.

[14] Dhandapani H, Siddiqui A, Karadkar S, Tayalia P. In vitro 3D spheroid model preserves tumor microenvironment of hot and cold breast cancer subtypes. Adv Healthc Mater. 2023; 12(21): e2300164.

[15] Abril-Rodriguez G, Torrejon DY, Liu W, et al. PAK4 inhibition improves PD-1 blockade immunotherapy. Nat Cancer. 2020; 1(1): 46-58.

[16] Wang JL, Hua SN, Bao HJ, Yuan J, Zhao Y, Chen S. Pyroptosis and inflammasomes in cancer and inflammation. MedComm. 2023; 4(5): e374.

[17] Jin X, Ma Y, Liu D, Huang Y. Role of pyroptosis in the pathogenesis and treatment of diseases. MedComm. 2023; 4(3): e249.

[18] Xiao Y, Zhang T, Ma X, et al. Microenvironment-responsive prodrug-induced pyroptosis boosts cancer immunotherapy. Adv Sci (Weinh). 2021; 8(24): e2101840.

[19] Gielecińska A, Kciuk M, Yahya EB, Ainane T, Mujwar S, Kontek R. Apoptosis, necroptosis, and pyroptosis as alternative cell death pathways induced by chemotherapeutic agents? Biochim Biophys Acta Rev Cancer. 2023; 1878(6): 189024.

[20] Zhang CC, Li CG, Wang YF, et al. Chemotherapeutic paclitaxel and cisplatin differentially induce pyroptosis in A549 lung cancer cells via caspase-3/GSDME activation. Apoptosis. 2019; 24(3–4): 312-325.

[21] Wu J, Wu Y, Chen S, et al. PARP1-stabilised FOXQ1 promotes ovarian cancer progression by activating the LAMB3/WNT/β-catenin signalling pathway. Oncogene. 2024; 43(12): 866-883.

[22] Gonzalez-Villasana V, Fuentes-Mattei E, Ivan C, et al. Rac1/Pak1/p38/MMP-2 axis regulates angiogenesis in ovarian cancer. Clin Cancer Res. 2015; 21(9): 2127-2137.

[23] Fang D, Chen H, Zhu JY, et al. Epithelial-mesenchymal transition of ovarian cancer cells is sustained by Rac1 through simultaneous activation of MEK1/2 and Src signaling pathways. Oncogene. 2017; 36(11): 1546-1558.

[24] Mosca L, Ilari A, Fazi F, Assaraf YG, Colotti G. Taxanes in cancer treatment: activity, chemoresistance and its overcoming. Drug Resist Updat. 2021; 54: 100742.

[25] Su L, Chen Y, Huang C, et al. Targeting Src reactivates pyroptosis to reverse chemoresistance in lung and pancreatic cancer models. Sci Transl Med. 2023; 15(678): eabl7895.

[26] Brown AL, Foster KL, Lupo PJ, et al. DNA methylation of a novel PAK4 locus influences ototoxicity susceptibility following cisplatin and radiation therapy for pediatric embryonal tumors. Neuro Oncol. 2017; 19(10): 1372-1379.

[27] Yan C, Niu Y, Li F, Zhao W, Ma L. System analysis based on the pyroptosis-related genes identifies GSDMC as a novel therapy target for pancreatic adenocarcinoma. J Transl Med. 2022; 20(1): 455.

[28] Dasgupta A, Sierra L, Tsang SV, et al. Targeting PAK4 inhibits Ras-mediated signaling and multiple oncogenic pathways in high-risk rhabdomyosarcoma. Cancer Res. 2021; 81(1): 199-212.

[29] Gao W, Wang X, Zhou Y, Wang X, Yu Y. Autophagy, ferroptosis, pyroptosis, and necroptosis in tumor immunotherapy. Signal Transduct Target Ther. 2022; 7(1): 196.

[30] Chen L, Long X, Duan S, et al. CSRP2 suppresses colorectal cancer progression via p130Cas/Rac1 axis-meditated ERK, PAK, and HIPPO signaling pathways. Theranostics. 2020; 10(24): 11063-11079.

[31] Sun X, Liu S, Wang J, et al. Annexin A5 regulates hepatocarcinoma malignancy via CRKI/II-DOCK180-RAC1 integrin and MEK-ERK pathways. Cell Death Dis. 2018; 9(6): 637.

[32] Faria M, Domingues R, Bugalho MJ, Matos P, Silva AL. MAPK inhibition requires active RAC1 signaling to effectively improve iodide uptake by thyroid follicular cells. Cancers (Basel). 2021; 13(22): 5861.

[33] Zhang X, Li Q, Wang L, Liu ZJ, Zhong Y. Active protection: learning-activated Raf/MAPK activity protects labile memory from Rac1-independent forgetting. Neuron. 2018; 98(1): 142-155. e144.

[34] Huang S, Deng W, Wang P, et al. Fermitin family member 2 promotes melanoma progression by enhancing the binding of p-alpha-Pix to Rac1 to activate the MAPK pathway. Oncogene. 2021; 40(37): 5626-5638.

[35] Pandey D, Goyal P, Dwivedi S, Siess W. Unraveling a novel Rac1-mediated signaling pathway that regulates cofilin dephosphorylation and secretion in thrombin-stimulated platelets. Blood. 2009; 114(2): 415-424.

[36] Wang Y, Guo F. Group I PAKs in myelin formation and repair of the central nervous system: what, when, and how. Biol Rev Camb Philos Soc. 2022; 97(2): 615-639.

[37] Trerotola M, Jernigan DL, Liu Q, Siddiqui J, Fatatis A, Languino LR. Trop-2 promotes prostate cancer metastasis by modulating β(1) integrin functions. Cancer Res. 2013; 73(10): 3155-3167.

[38] Kudo K, Greer YE, Yoshida T, et al. Dual-inhibition of NAMPT and PAK4 induces anti-tumor effects in 3D-spheroids model of platinum-resistant ovarian cancer. Cancer Gene Ther. 2024; 31(5): 721-735.

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2024 The Author(s). MedComm published by Sichuan International Medical Exchange & Promotion Association (SCIMEA) and John Wiley & Sons Australia, Ltd.