Effect of PLGA Nanoparticle-Mediated Delivery of miRNA 503 on The Apoptosis of Ovarian Endometriosis Cells

Document Type : Original Article

Authors

1 Endometriosis Research Center, Iran University of Medical Sciences, Tehran, Iran

2 Pars Advanced and Minimally Invasive Medical Manners Research Center, Pars Hospital, Iran University of Medical Sciences, Tehran, Iran

3 Endometriosis Research Center, Iran University of Medical Sciences, Tehran, Iran.

4 Neurosciences Research Center, Iran University of Medical Sciences, Tehran, Iran

5 Antimicrobial Resistance Research Center, Institute of Immunology and Infectious Diseases, Iran University of Medical Sciences, Tehran, Iran

6 Air Pollution Research Center, Iran University of Medical Sciences, Tehran, Iran

Abstract

Objective:
One of the challenges in gene therapy is the transfer of the gene to the target cell. MicroRNAs (miRNAs)
regulate gene expression after transcription by binding directly to the messenger and play a vital role in cell behaviors
and the pathogenesis of some diseases. This study was aimed at developing poly (lactic-co-glycolic acid) (PLGA)-
based nanoparticles (NPs) for gene delivery to endometriotic cyst stromal cells (ECSCs).

Materials and Methods:
In this experimental study, endometriosis cells were isolated from women with severe
endometriosis (DIE) and digested by the enzymatic method (40 μg/ml DNAase I and 300 μg/ml collagenase type 3).
PLGA-based NPs were synthesized and characterized. The size of sole PLGA NPs and PLGA/miRNA were 60 ± 4 nm
and 70 ± 5.1 nm respectively. Poly lactic-co-glycolic-based NPs were used as vector carriers for miRNA 503 transfection
in endometriosis cells. The cells were divided into the five groups of control and four doses (25, 50, 75, and 100 μm) of
miRNA 503/PLGA at 12, 24, 48, and 72 hours. Viability and apoptosis were evaluated by the MTT assay and Annexin
Kits. Data were analyzed by one-way analysis of variance.

Results:
The results show that the size of PLGA/miRNA complex with dynamic light scattering (DLS) was 70 ± 5.1
nm and zeta potential values of the PLGA/PEI/miRNA complexes were 27.9 mV. Based on the MTT assay results,
the optimal dose of miRNA 503/PLGA was 75 μm, at which the viability of ECSCs was 52.6% ± 1.2 (P≤0.001), and
the optimal time was 48 hours. The apoptotic rates of ECSCs treated with PLGA/miRNA503 (34.75 ± 4.9%) were
significantly higher than those of ECSCs treated with PLGA alone (3.35 ± 2.58%, P≤0.01).

Conclusion:
Cell death increased with increasing the concentration of miRNA; thus, it can be suggested as a treatment
for endometriosis.

Keywords


  1. Cranney R, Condous G, Reid S. An update on the diagnosis, surgical management, and fertility outcomes for women with endometrioma. Acta Obstet Gynecol Scand. 2017; 96(6): 633-643.
  2. Ding S, Guo X, Zhu L, Wang J, Li T, Yu Q, et al. Macrophagederived netrin-1 contributes to endometriosis-associated pain. Ann Transl Med. 2021; 9(1): 29.
  3. Orazov MR, Radzinsky VY, Ivanov II, Khamoshina MB, Shustova VB. Oocyte quality in women with infertility associated endometriosis. Gynecol Endocrinol. 2019; 35 Suppl 1: 24-26.
  4. Ronchi A, Grauso F, Marino FZ, Quagliariello V, Maurea N, Facchini G, et al. Endocannabinoid system expression in ovarian epithelial tumors according to the dualistic model of ovarian carcinogenesis. Eur Rev Med Pharmacol Sci. 2021; 25(14): 4678- 4686.
  5. Tanaka K, Mayne L, Khalil A, Baartz D, Eriksson L, Mortlock SA, et al. The role of the endocannabinoid system in aetiopathogenesis of endometriosis: a potential therapeutic target. Eur J Obstet Gynecol Reprod Biol. 2020; 244: 87-94.
  6. Guo SW. Endometriosis and ovarian cancer: potential benefits and harms of screening and risk-reducing surgery. Fertil Steril. 2015; 104(4): 813-830.
  7. Schleedoorn MJ, Nelen WL, Dunselman GA, Vermeulen N; EndoKey Group. Selection of key recommendations for the management of women with endometriosis by an international panel of patients and professionals. Hum Reprod. 2016; 31(6): 1208- 1218.
  8. Mechsner S. Endometriosis: an often unrecognized pain disorder. Schmerz. 2016; 30(5): 477-490.
  9. Raja MHR, Farooqui N, Zuberi N, Ashraf M, Azhar A, Baig R, et al. Endometriosis, infertility and MicroRNA’s: a review. J Gynecol Obstet Hum Reprod. 2021; 50(9): 102157.
  10. Marí-Alexandre J, Barceló-Molina M, Belmonte-López E, García-Oms J, Estellés A, Braza-Boïls A, et al. Micro-RNA profile and proteins in peritoneal fluid from women with endometriosis: their relationship with sterility. Fertil Steril. 2018; 109(4): 675-684. e2.
  11. Sbracia M, Valeri C, Antonini G, Biagiotti G, Pacchiarotti A, Pacchiarotti A. Fas and Fas-ligand in eutopic and ectopic endometrium of women with endometriosis: the possible immune privilege of ectopic endometrium. Reprod Sci. 2016; 23(1): 81-86.
  12. Bai Z, Wei J, Yu C, Han X, Qin X, Zhang C, et al. Non-viral nanocarriers for intracellular delivery of microRNA therapeutics. J Mater Chem B. 2019; 7(8): 1209-1225.
  13. Ramezani M, Ebrahimian M, Hashemi M. Current strategies in the modification of PLGA-based gene delivery system. Curr Med Chem. 2017; 24(7): 728-739.
  14. Marí-Alexandre J, Sánchez-Izquierdo D, Gilabert-Estellés J, Barceló-Molina M, Braza-Boïls A, Sandoval J. miRNAs regulation and its role as biomarkers in endometriosis. Int J Mol Sci. 2016; 17(1): 93.
  15. Hirakawa T, Nasu K, Abe W, Aoyagi Y, Okamoto M, Kai K, et al. miR-503, a microRNA epigenetically repressed in endometriosis, induces apoptosis and cell-cycle arrest and inhibits cell proliferation, angiogenesis, and contractility of human ovarian endometriotic stromal cells. Hum Reprod. 2016; 31(11): 2587- 2597.
  16. Brueggmann D, Templeman C, Starzinski-Powitz A, Rao NP, Gayther SA, Lawrenson K. Novel three-dimensional in vitro models of ovarian endometriosis. J Ovarian Res. 2014; 7: 17.
  17. Konrad L, Kortum J, Nabham R, Gronbach J, Dietze R, Oehmke F, et al. Composition of the stroma in the human endometrium and endometriosis. Reprod Sci. 2018; 25(7): 1106-1115.
  18. Swider E, Koshkina O, Tel J, Cruz LJ, de Vries IJM, Srinivas M. Customizing poly(lactic-co-glycolic acid) particles for biomedical applications. Acta Biomater. 2018; 73: 38-51.
  19. Shams A, Shabani R, Najafi M, Karimi M, Pirhajati V, Asghari Jafarabadi M, et al. Therole of MicroRNA 143 and MicroRNA 206 in the regulation of apoptosis in mouse lukemia cancer cells and spermatogonial cells. Cell J. 2021; 23(5): 544-551.
  20. Crowley LC, Marfell BJ, Scott AP, Waterhouse NJ. Quantitation of apoptosis and necrosis by annexin V binding, propidium iodide uptake, and flow cytometry. Cold Spring Harb Protoc. 2016; 2016(11).
  21. Jafarabadi M, Salehnia M, Sadafi R. Evaluation of two endometriosis models by transplantation of human endometrial tissue fragments and human endometrial mesenchymal cells. Int J Reprod Biomed. 2017; 15(1): 21-32.
  22. Rai R, Alwani S, Badea I. Polymeric nanoparticles in gene therapy: New avenues of design and optimization for delivery applications. Polymers (Basel). 2019; 11(4): 745.
  23. Delbandi AA, Mahmoudi M, Shervin A, Heidari S, Kolahdouz- Mohammadi R, Zarnani AH. Evaluation of apoptosis and angiogenesis in ectopic and eutopic stromal cells of patients with endometriosis compared to non-endometriotic controls. BMC Womens Health. 2020; 20(1): 3.
  24. Dahiya A, Sebastian A, Thomas A, George R, Thomas V, Peedicayil A. Endometriosis and malignancy: the intriguing relationship. Int J Gynaecol Obstet. 2021; 155(1): 72-78.
  25. Quagliariello V, Berretta M, Buccolo S, Iovine M, Paccone A, Cavalcanti E, et al. Polydatin reduces cardiotoxicity and enhances the anticancer effects of sunitinib by decreasing pro-oxidative stress, pro-inflammatory cytokines, and NLRP3 inflammasome expression. Front Oncol. 2021; 11: 680758.
  26. Riley KA, Benton AS, Deimling TA, Kunselman AR, Harkins GJ. Surgical excision versus ablation for superficial endometriosisassociated pain: a randomized controlled trial. J Minim Invasive Gynecol. 2019; 26(1): 71-77.
  27. Mehrdad Moghimi M, Bahram Moazzami M. Comparing the efficacy of surgery and medical therapy for pain management in endometriosis: a systematic review and meta-analysis. Pain Physician. 2017; 20: 185-95.
  28. Samartzis EP, Fink D, Stucki M, Imesch P. Doxycycline reduces MMP-2 activity and inhibits invasion of 12Z epithelial endometriotic cells as well as MMP-2 and-9 activity in primary endometriotic stromal cells in vitro. Reprod Biol Endocrinol. 2019; 17(1): 38.
  29. Eslahi N, Shakeri-Zadeh A, Ashtari K, Pirhajati-Mahabadi V, Tohidi Moghadam T, Shabani R, et al. In vitro cytotoxicity of folatesilica- gold nanorods on mouse acute lymphoblastic leukemia and spermatogonial cells. Cell J. 2019; 21(1): 14-26.
  30. Chaudhury K, Babu KN, Singh AK, Das S, Kumar A, Seal S. Mitigation of endometriosis using regenerative cerium oxide nanoparticles. Nanomedicine. 2013; 9(3): 439-448.
  31. de Almeida Borges VR, Tavares MR, da Silva JH, Tajber L, Boylan F, Ribeiro AF, et al. Development and characterization of poly(lactic-co-glycolic) acid nanoparticles loaded with copaiba oleoresin. Pharm Dev Technol. 2018; 23(4): 343-350.
  32. Chenthamara D, Subramaniam S, Ramakrishnan SG, Krishnaswamy S, Essa MM, Lin F-H, et al. Therapeutic efficacy of nanoparticles and routes of administration. Biomater Res. 2019; 23: 20.
  33. Berthet M, Gauthier Y, Lacroix C, Verrier B, Monge C. Nanoparticle- based dressing: the future of wound treatment? Trends Biotechnol. 2017; 35(8): 770-784.
  34. Shabani R, Ashjari M, Ashtari K, Izadyar F, Behnam B, Khoei S, et al. Elimination of mouse tumor cells from neonate spermatogonial cells utilizing cisplatin-entrapped folic acid-conjugated poly (lactic-co-glycolic acid) nanoparticles in vitro. Int J Nanomedicine. 2018; 13: 2943-2954.
  35. Singh AK, Chakravarty B, Chaudhury K. Nanoparticle-assisted combinatorial therapy for effective treatment of endometriosis. J Biomed Nanotechnol. 2015; 11(5): 789-804.
  36. Guo X, Li W, Zhou J, Hou W, Wen X, Zhang H, et al. Specific photothermal ablation therapy of endometriosis by targeting delivery of gold nanospheres. Small. 2017; 13(15): 1603270.
  37. Li J, He Y, Sun W, Luo Y, Cai H, Pan Y, et al. Hyaluronic acidmodified hydrothermally synthesized iron oxide nanoparticles for targeted tumor MR imaging. Biomaterials. 2014; 35(11): 3666-3677.
  38. Braza-Boïls A, Marí-Alexandre J, Gilabert J, Sánchez-Izquierdo D, España F, Estellés A, et al. MicroRNA expression profile in endometriosis: its relation to angiogenesis and fibrinolytic factors. Hum Reprod. 2014; 29(5): 978-988.
  39. Shams A, Shabani R, Asgari H, Karimi M, Najafi M, Asghari- Jafarabadi M, et al. In vitro elimination of EL4 cancer cells from spermatogonia stem cells by miRNA-143-and 206-loaded folic acid-conjugated PLGA nanoparticles. Nanomedicine. 2022; 17(8): 531-545.
  40. Wang F, Salvati A, Boya P. Lysosome-dependent cell death and deregulated autophagy induced by amine-modified polystyrene nanoparticles. Open Biol. 2018; 8(4): 170271.