Journal of Reproduction & Infertility

Journal of Reproduction & Infertility

Altered Expression of GABPB1-IT1 and SLC9A3-AS1 Long Non-Coding RNAs in Recurrent Implantation Failure

Authors
1 Human Genetics Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran
2 Department of Genetics, Tabriz University of Medical Sciences, Tabriz, Iran
3 Student Research Committee, School of Medicine, Iran University of Medical Sciences (IUMS), Tehran, Iran
4 Vali-E-Asr Reproductive Health Research Center, Family Health Research Institute, Tehran University of Medical Sciences, Tehran, Iran
Abstract
Background: Recurrent implantation failure (RIF) impairs assisted reproductive technology (ART) success, though its mechanisms remain unclear. Studies have implicated altered miRNA expression in the plasma and endometrium of patients with RIF. In this study, long non-coding RNA (lncRNA) expression in plasma was investigated, using RIF-specific lncRNA-miRNA-mRNA network as guide. Methods: This study included 30 women with RIF and 30 age-matched controls (Not RIF). RIF miRNA RNA-seq data (GSE108966) was downloaded from the Gene Expression Omnibus (GEO) database and used to identify differentially expressed miRNAs. Next, miRNet 2.0 database was used to integrate the latest miRNA-mRNA interactions; also, the linkages between lncRNA and miRNA were identified and the networks created. Plasma samples were collected from participants during the implantation window. Cell-free RNA was extracted from 500 µl of plasma using the TRIzol method, and statistical analyses were performed using Prism version 8.0. Results: Based on a lncRNA-miRNA-mRNA network analysis, two lncRNAs, including GABPB1-IT1 and SLC9A3-AS1, were selected for expression analysis in plasma. Notably, GABPB1-IT1 and SLC9A3-AS1 were significantly downregulated in RIF samples compared to controls. Conclusion: Our findings show that GABPB1-IT1 and SLC9A3-AS1 are significantly downregulated in the plasma of women with recurrent implantation failure. These findings suggest their potential involvement in RIF and warrant further studies to elucidate their biological functions and evaluate their utility as non-invasive biomarkers.
Keywords

  1. Coughlan C, Ledger W, Wang Q, Liu F, Demirol A, Gurgan T, et al. Recurrent implantation failure: definition and management. Reprod Biomed Online. 2014;28(1):14-38.
  2. Busnelli A, Reschini M, Cardellicchio L, Vegetti W, Somigliana E, Vercellini P. How common is real repeated implantation failure? an indirect estimate of the prevalence. Reprod Biomed Online. 2020;40(1):91-7.
  3. Cimadomo D, Craciunas L, Vermeulen N, Vomstein K, Toth B. Definition, diagnostic and therapeutic options in recurrent implantation failure: an international survey of clinicians and embryologists. Hum Reprod. 2021;36(2):305-17.
  4. Mojarrad M, Hassanzadeh-Nazarabadi M, Tafazoli N. Polymorphism of genes and implantation failure. Int J Mol Cell Med. 2013;2(1):1-8.
  5. Craciunas L, Gallos I, Chu J, Bourne T, Quenby S, Brosens JJ, et al. Conventional and modern markers of endometrial receptivity: a systematic review and meta-analysis. Hum Reprod Update. 2019;25(2):202-23.
  6. Fazli F, Khanlarzadeh E, Pilehvari Sh. The impact of l-arginine on uterine artery resistance and pregnancy outcomes in frozen embryo transfer for IVF candidates with recurrent implantation failure: a clinical trial. J Reprod Infertil. 2025;26(1):19-27.
  7. Zaki-Dizaji M, Saeedinia M, Derogar P, Jamshidi B, Masoumi M, Heidary Z. hsa_circ_0004121 and hsa_circ_0030162 differentially expressed in plasma of patients with recurrent implantation failure. Rep Biochem Mol Biol. 2024;13(3):428-37.
  8. Nemeth K, Bayraktar R, Ferracin M, Calin GA. Non-coding RNAs in disease: from mechanisms to therapeutics. Nat Rev Genet. 2024;25(3):211-32.
  9. Zahir M, Tavakoli B, Zaki-Dizaji M, Hantoushzadeh S, Majidi Zolbin M. Non-coding RNAs in recurrent implantation failure. Clin Chim Acta. 2024;553:117731.
  10. Azhari F, Pence S, Hosseini MK, Balci BK, Cevik N, Bastu E, et al. The role of the serum exosomal and endometrial microRNAs in recurrent implantation failure. J Matern Fetal Neonatal Med. 2022;35(5):815-25.
  11. Zeng H, Fu Y, Shen L, Quan S. MicroRNA signatures in plasma and plasma exosome during window of implantation for implantation failure following in-vitro fertilization and embryo transfer. Reprod Biol Endocrinol. 2021;19(1):180.
  12. Chen P, Li T, Guo Y, Jia L, Wang Y, Fang C. Construction of circulating microRNAs-based non-invasive prediction models of recurrent implantation failure by network analysis. Front Genet. 2021;12:712150.
  13. Yang Q, Gu WW, Gu Y, Yan NN, Mao YY, Zhen XX, et al. Association of the peripheral blood levels of circulating microRNAs with both recurrent miscarriage and the outcomes of embryo transfer in an in vitro fertilization process. J Transl Med. 2018;16(1):186.
  14. Freis A, Keller A, Ludwig N, Meese E, Jauckus J, Rehnitz J, et al. Altered miRNA-profile dependent on ART outcome in early pregnancy targets Wnt-pathway. Reproduction. 2017;154(6):799-805.
  15. Huang J, Song N, Xia L, Tian L, Tan J, Chen Q, et al. Construction of lncRNA-related competing endogenous RNA network and identification of hub genes in recurrent implantation failure. Reprod Biol Endocrinol. 2021;19(1):108.
  16. Feng C, Shen JM, Lv PP, Jin M, Wang LQ, Rao JP, et al. Construction of implantation failure related lncRNA-mRNA network and identification of lncRNA biomarkers for predicting endometrial receptivity. Int J Biol Sci. 2018;14(10):1361-77.
  17. Zhao H, Hu S, Qi J, Wang Y, Ding Y, Zhu Q, et al. Increased expression of HOXA11-AS attenuates endometrial decidualization in recurrent implantation failure patients. Mol Ther. 2022;30(4):1706-20.
  18. Chen MY, Liao GD, Zhou B, Kang LN, He YM, Li SW. Genome-wide profiling of long noncoding RNA expression patterns in women with repeated implantation failure by RNA sequencing. Reprod Sci. 2019;26(1):18-25.
  19. Ahmadi M, Pashangzadeh S, Moraghebi M, Sabetian S, Shekari M, Eini F, et al. Construction of circRNA-miRNA-mRNA network in the pathogenesis of recurrent implantation failure using integrated bioinformatics study. J Cell Mol Med. 2022;26(6):1853-64.
  20. Zhao H, Chen L, Shan Y, Chen G, Chu Y, Dai H, et al. Hsa_circ_0038383-mediated competitive endogenous RNA network in recurrent implantation failure. Aging (Albany NY). 2021;13(4):6076-90.
  21. Zhou T, Ni T, Li Y, Zhang Q, Yan J, Chen ZJ. circFAM120A participates in repeated implantation failure by regulating decidualization via the miR-29/ABHD5 axis. FASEB J. 2021;35(9):e21872.
  22. Zhao F, Guo Y, Shi Z, Wu M, Lv Y, Song W. hsa_circ_001946 elevates HOXA10 expression and promotes the development of endometrial receptivity via sponging miR-135b. Diagn Pathol. 2021;16(1):44.
  23. Ni T, Zhang Q, Li Y, Huang C, Zhou T, Yan J, et al. CircSTK40 contributes to recurrent implantation failure via modulating the HSP90/AKT/FOXO1 axis. Mol Ther Nucleic Acids. 2021;26:208-21.
  24. Luo J, Zhu L, Zhou N, Zhang Y, Zhang L, Zhang R. Construction of circular RNA-microRNA-messenger RNA regulatory network of recurrent implantation failure to explore its potential pathogenesis. Front Genet. 2020;11:627459.
  25. Liu L, Li L, Ma X, Yue F, Wang Y, Wang L, et al. Altered circular RNA expression in patients with repeated implantation failure. Cell Physiol Biochem. 2017;44(1):303-13.
  26. Mattick JS, Amaral PP, Carninci P, Carpenter S, Chang HY, Chen LL, et al. Long non-coding RNAs: definitions, functions, challenges and recommendations. Nat Rev Mol Cell Biol. 2023;24(6):430-47.
  27. Beylerli O, Gareev I, Sufianov A, Ilyasova T, Guang Y. Long noncoding RNAs as promising biomarkers in cancer. Noncoding RNA Res. 2022;7(2):66-70.
  28. Razzaghi H, Heiat M, Khoncheh A, Abyazi MA, Zaki-Dizaji M. Platelet-derived circRNAs hsa_circ_0004771 and hsa_circ_0019120 differentially expressed in colorectal cancer and polyps. Rep Biochem Mol Biol. 2024;13(3):368-76.
  29. Rekker K, Altmäe S, Suhorutshenko M, Peters M, Martinez-Blanch JF, Codoñer FM, et al. A two-cohort RNA-seq study reveals changes in endometrial and blood miRNome in fertile and infertile women. Genes (Basel). 2018;9(12):574.
  30. Andrews S. FastQC: a quality control tool for high throughput sequence data. Cambridge (UK): Babraham Bioinformatics; 2010.
  31. Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for illumina sequence data. Bioinformatics. 2014;30(15):2114-20.
  32. Friedländer MR, Mackowiak SD, Li N, Chen W, Rajewsky N. miRDeep2 accurately identifies known and hundreds of novel microRNA genes in seven animal clades. Nucleic Acids Res. 2012;40(1):37-52.
  33. Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 2014;15(12):550.
  34. Chang L, Zhou G, Soufan O, Xia J. miRNet 2.0: network-based visual analytics for miRNA functional analysis and systems biology. Nucleic Acids Res. 2020;48(W1):W244-51.
  35. Ruijter JM, Ramakers C, Hoogaars WM, Karlen Y, Bakker O, van den Hoff MJ, et al. Amplification efficiency: linking baseline and bias in the analysis of quantitative PCR data. Nucleic Acids Res. 2009;37(6):e45.
  36. Feng F, Zhang R, Long L. LncRNA GABPB1-IT1 Is upregulated in ischemia-induced acute kidney injury and downregulates miR-204-5p to promote hypoxia-induced human renal proximal tubular epithelial cell apoptosis. Kidney Blood Press Res. 2024;49(1):480-9.
  37. Luo C, Zhang J, Bo L, Wei L, Yang G, Gao S, et al. Construction of a ceRNA-based lncRNA-mRNA network to identify functional lncRNAs in premature ovarian insufficiency. Front Genet. 2022;13:956805.
  38. Bai Y, Qu Y, Wu Z, Ren Y, Cheng Z, Lu Y, et al. Absolute quantification and analysis of extracellular vesicle lncRNAs from the peripheral blood of patients with lung cancer based on multi-colour fluorescence chip-based digital PCR. Biosens Bioelectron. 2019;142:111523.
  39. Li J, Jing J, Liu J, Zhang D, Zhang L, Xie G. Integration of transcriptome and DNA methylation reveals the mechanism of cilia-related genes in recurrent miscarriage. Sci Rep. 2026;16(1):21324.
  40. Xie J, Xie G, Chen Q, Xu Z, Bai W, Chen M. Identification of a novel lncRNA GABPB1-IT1 that is downregulated and predicts a poor prognosis in non-small cell lung cancer. Oncol Lett. 2019;18(1):838-45.
  41. Li B, Wei Y, Ge Q, Duan Y, Guo L. lncRNA GABPB1 intronic transcript 1 upregulates pigment epithelium-derived factor via miR-93 to suppress cell proliferation in hepatocellular carcinoma. Oncol Lett. 2021;21(4):260.
  42. Wang T, Cao C, Fan Y, Xu J, Hua T, Ding J, et al. GABPB1 plays a cancer-promoting role in non-small cell lung cancer. Discov Oncol. 2024;15(1):72.
  43. Tan C, Du H, Wang Y, Zhao J, Cheng X, Lan H. LncRNA GABPB1-IT1 inhibits the tumorigenesis of renal cancer via the miR-21/PTEN axis. J Biochem Mol Toxicol. 2023;37(4):e23288.
  44. Huang Y, Li L, Kang Z, Luo H, Lin X, Zhao S, et al. Prognostic model associated with necroptosis in colorectal cancer based on transcriptomic analysis and experimental validation. Front Biosci (Landmark Ed). 2024;29(3):98.
  45. Dalla Torre M, Pittari D, Boletta A, Cassina L, Sitia R, Anelli T. Mitochondria remodeling during endometrial stromal cell decidualization. Life Sci Alliance. 2024;7(12):e202402627.
  46. Yang ZF, Drumea K, Mott S, Wang J, Rosmarin AG. GABP transcription factor (nuclear respiratory factor 2) is required for mitochondrial biogenesis. Mol Cell Biol. 2014;34(17):3194-201.
  47. Li J, Li D, Zhang X, Li C, Zhu F. Long noncoding RNA SLC9A3 AS1 increases E2F6 expression by sponging microRNA 486 5p and thus facilitates the oncogenesis of nasopharyngeal carcinoma. Oncol Rep. 2021;46(2):165.
  48. Huang X, Huang M, Chen M, Chen X. lncRNA SLC9A3-AS1 promotes oncogenesis of NSCLC via sponging microRNA-760 and may serve as a prognosis predictor of NSCLC patients. Cancer Manag Res. 2022;14:1087-98.
  49. Ye C, Qin S, Qiu S, Zhao L, Miao J, Chen Y, et al. A lncRNA-immune checkpoint-related gene signature predicts metastasis-free survival in prostate adenocarcinoma. Transl Androl Urol. 2022;11(12):1691-705.
  50. Zhao L, Zhang H, Ren P, Sun X. LncRNA SLC9A3-AS1 knockdown increases the sensitivity of liver cancer cell to triptolide by regulating miR-449b-5p-mediated glycolysis. Biotechnol Genet Eng Rev. 2024;40(2):1389-405.
  51. Zhou T, Nguyen S, Wu J, He B, Feng Q. LncRNA LOC730101 promotes darolutamide resistance in prostate cancer by suppressing miR-1-3p. Cancers (Basel). 2024;16(14):2594.
  52. Chen KC, Chang ML, Lin CS, Rajneesh CP, Liao CH, You WC, et al. Insight into SLC9A3 deficiency-mediated micturition dysfunction caused by electrolyte imbalance. Biomed Pharmacother. 2023;158:114155.
  53. Ruan YC, Chen H, Chan HC. Ion channels in the endometrium: regulation of endometrial receptivity and embryo implantation. Hum Reprod Update. 2014;20(4):517-29.