Abstract
Background
Preeclampsia (PE) is one of the most serious pregnancy complications with unknown pathogenesis. Emerging evidence has demonstrated that Fms-related tyrosine kinase 1 (FLT1) is highly involved in PE development. As a pseudogene of FLT1, FLT1P1 increased in PE samples. However, its functions remain largely unknown.
Methods and results
In this study, co-expression analysis was performed to identify the potential target genes of FTL1P1. Then chromatin isolation using RNA purification (ChIRP) method was employed to explore the interactomes of FLT1P1, including interacting with DNA fragments and proteins. We found that in PE samples, both FLT1P1 and FLT1 were highly expressed and closely correlated. ChIRP-protein data revealed that FLT1P1 interacts with translation- and transcription-related proteins, including 4 transcription factors (TFs). ChIRP-DNA analysis revealed that FLT1P1 preferentially interacted with DNA fragments downstream of transcription start sites (TSSs). Functional analysis of its interacting genes revealed that they were enriched in transcriptional regulation and apoptosis-related pathways. Twenty-six TFs, including CREB1 and SRF, were extracted from the potential FLT1P1-interacting gene sets and were potential targets of FLT1P1. CREB1 could bind to FLT1 promoter, and was negatively correlated with FLT1 at the expression level, making it a potential regulator of FLT1.
Conclusions
Our study extensively investigated the interactome profiles of FLT1P1, especially the prompter region of TF gene CREB1, and revealed the potential molecular regulatory mechanisms of FLT1 expression in PE samples. Our results provide a novel view of PE pathogenesis, and suggest that FLT1P1 could serve as a potential therapeutic target in PE diagnosis and treatment.
Similar content being viewed by others
Data availability
The datasets analyzed for this study can be found in the Gene Expression Omnibus (GEO) database with accession number GSE183012 https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE183012.
References
Rana S, Lemoine E, Granger JP, Karumanchi SA (2019) Preeclampsia: pathophysiology, challenges, and perspectives. Circ Res 124:1094–1112
Duckitt K, Harrington D (2005) Risk factors for pre-eclampsia at antenatal booking: systematic review of controlled studies. BMJ 330:565
Redman CW, Sargent IL (2005) Latest advances in understanding preeclampsia. Science 308:1592–1594
Romero R, Chaiworapongsa T (2013) Preeclampsia: a link between trophoblast dysregulation and an antiangiogenic state. J Clin Invest 123:2775–2777
Gray KJ, Kovacheva VP, Mirzakhani H, Bjonnes AC, Almoguera B, DeWan AT, Triche EW, Saftlas AF, Hoh J, Bodian DL (2018) Gene-centric analysis of preeclampsia identifies maternal association at PLEKHG1. Hypertension 72:408–416
McGinnis, R., Steinthorsdottir, V., Williams, N. O., Thorleifsson, G., Shooter, S., Hjartardottir, S., Bumpstead, S., Stefansdottir, L., Hildyard, L., Sigurdsson, J. K., Kemp, J. P., Silva, G. B., Thomsen, L. C. V., Jaaskelainen, T., Kajantie, E., Chappell, S., Kalsheker, N., Moffett, A., Hiby, S., Lee, W. K., Padmanabhan, S., Simpson, N. A. B., Dolby, V. A., Staines-Urias, E., Engel, S. M., Haugan, A., Trogstad, L., Svyatova, G., Zakhidova, N., Najmutdinova, D., Consortium, F., Consortium, G., Dominiczak, A. F., Gjessing, H. K., Casas, J. P., Dudbridge, F., Walker, J. J., Pipkin, F. B., Thorsteinsdottir, U., Geirsson, R. T., Lawlor, D. A., Iversen, A. C., Magnus, P., Laivuori, H., Stefansson, K. & Morgan, L. (2017) Variants in the fetal genome near FLT1 are associated with risk of preeclampsia, Nat Genet. 49, 1255-1260.
Cnattingius S, Reilly M, Pawitan Y, Lichtenstein P (2004) Maternal and fetal genetic factors account for most of familial aggregation of preeclampsia: a population-based Swedish cohort study. Am J Med Genet A 130A:365–371
Maynard SE, Karumanchi SA (2011) Angiogenic factors and preeclampsia. Semin Nephrol. https://doi.org/10.1016/j.semnephrol.2010.10.004
Levine RJ, Maynard SE, Qian C, Lim K-H, England LJ, Yu KF, Schisterman EF, Thadhani R, Sachs BP, Epstein FH (2004) Circulating angiogenic factors and the risk of preeclampsia. N Engl J Med 350:672–683
Maynard SE, Min JY, Merchan J, Lim KH, Li J, Mondal S, Libermann TA, Morgan JP, Sellke FW, Stillman IE, Epstein FH, Sukhatme VP, Karumanchi SA (2003) Excess placental soluble fms-like tyrosine kinase 1 (sFlt1) may contribute to endothelial dysfunction, hypertension, and proteinuria in preeclampsia. J Clin Invest 111:649–658
Hod T, Cerdeira AS, Karumanchi SA (2015) Molecular mechanisms of preeclampsia. Cold Spring Harb Perspect Med 5:a023473
Mutter WP, Karumanchi SA (2008) Molecular mechanisms of preeclampsia. Microvasc Res 75:1–8
Robertson SA (2019) Preventing preeclampsia by silencing soluble Flt-1? N Engl J Med 380:1080–1082
Statello L, Guo CJ, Chen LL, Huarte M (2021) Gene regulation by long non-coding RNAs and its biological functions. Nat Rev Mol Cell Biol 22:96–118
Zhao H, Shi J, Zhang Y, Xie A, Yu L, Zhang C, Lei J, Xu H, Leng Z, Li T, Huang W, Lin S, Wang L, Xiao Y, Li X (2020) LncTarD: a manually-curated database of experimentally-supported functional lncRNA-target regulations in human diseases. Nucleic Acids Res 48:D118–D126
Song X, Luo X, Gao Q, Wang Y, Gao Q, Long W (2017) Dysregulation of LncRNAs in placenta and pathogenesis of preeclampsia. Curr Drug Targets 18:1165–1170
Liu C, Li H, Zhang Y, Ding H (2021) Long intergenic noncoding RNA 00473 promoting migration and invasion of trophoblastic cell line HTR-8/SVneo via regulating miR-424–5p-mediated wnt3a/beta-catenin signaling pathway. J Obstet Gynaecol Res. https://doi.org/10.1111/jog.14870
Zhang Y, He XY, Qin S, Mo HQ, Li X, Wu F, Zhang J, Li X, Mao L, Peng YQ, Guo YN, Lin Y, Tian FJ (2020) Upregulation of PUM1 expression in preeclampsia impairs trophoblast invasion by negatively regulating the expression of the lncRNA HOTAIR. Mol Ther 28:631–641
Gao Y, Guo X, Li Y, Sha W, She R (2019) The decreased lncRNA ZEB2-AS1 in pre-eclampsia controls the trophoblastic cell line HTR-8/SVneo’s invasive and migratory abilities via the miR-149/PGF axis. J Cell Biochem 120:17677–17686
He X, He Y, Xi B, Zheng J, Zeng X, Cai Q, Ouyang Y, Wang C, Zhou X, Huang H, Deng W, Xin S, Huang Q, Liu H (2013) LncRNAs expression in preeclampsia placenta reveals the potential role of LncRNAs contributing to preeclampsia pathogenesis. PLoS ONE 8:e81437
Chi Z, Sun Y, Yu Z, Zhou F, Wang H, Zhang M (2021) Pseudogene fms-related tyrosine kinase 1 pseudogene 1 (FLT1P1) cooperates with RNA binding protein dyskeratosis congenita 1 (DKC1) to restrain trophoblast cell proliferation and angiogenesis by targeting fms-related tyrosine kinase 1 (FLT1) in preeclampsia. Bioengineered 12:8885–8897
Poliseno L, Salmena L, Zhang J, Carver B, Haveman WJ, Pandolfi PP (2010) A coding-independent function of gene and pseudogene mRNAs regulates tumour biology. Nature 465:1033–1038
Harrison PM, Zheng D, Zhang Z, Carriero N, Gerstein M (2005) Transcribed processed pseudogenes in the human genome: an intermediate form of expressed retrosequence lacking protein-coding ability. Nucleic Acids Res 33:2374–2383
Rapicavoli NA, Qu K, Zhang J, Mikhail M, Laberge RM, Chang HY (2013) A mammalian pseudogene lncRNA at the interface of inflammation and anti-inflammatory therapeutics. Elife 2:e00762
Pei B, Sisu C, Frankish A, Howald C, Habegger L, Mu XJ, Harte R, Balasubramanian S, Tanzer A, Diekhans M (2012) The GENCODE pseudogene resource. Genome Biol 13:1–26
Pink RC, Wicks K, Caley DP, Punch EK, Jacobs L, Carter DRF (2011) Pseudogenes: pseudo-functional or key regulators in health and disease? RNA 17:792–798
Langmead B, Salzberg S (2012) Fast gapped-read alignment with Bowtie 2. Nat Methods 9:357–359
Zhang Y, Liu T, Meyer C, Eeckhoute J, Johnson D, Bernstein B, Nusbaum C, Myers R, Brown M, Li W, Liu X (2008) Model-based analysis of ChIP-Seq (MACS). Genome Biol 9:R137
Ramírez F, Ryan D, Grüning B, Bhardwaj V, Kilpert F, Richter A, Heyne S, Dündar F, Manke T (2016) deepTools2: a next generation web server for deep-sequencing data analysis. Nucleic Acids Res 44:W160–W165
Heinz S, Benner C, Spann N, Bertolino E, Lin Y, Laslo P, Cheng J, Murre C, Singh H, Glass C (2010) Simple combinations of lineage-determining transcription factors prime cis-regulatory elements required for macrophage and B cell identities. Mol Cell 38:576–589
Han H, Cho JW, Lee S, Yun A, Kim H, Bae D, Yang S, Kim CY, Lee M, Kim E, Lee S, Kang B, Jeong D, Kim Y, Jeon HN, Jung H, Nam S, Chung M, Kim JH, Lee I (2018) TRRUST v2: an expanded reference database of human and mouse transcriptional regulatory interactions. Nucleic Acids Res 46:D380–D386
Xie C, Mao X, Huang J, Ding Y, Wu J, Dong S, Kong L, Gao G, Li CY, Wei L (2011) KOBAS 2.0: a web server for annotation and identification of enriched pathways and diseases. Nucleic Acids Res 39:316–322
Turanov AA, Lo A, Hassler MR, Makris A, Ashar-Patel A, Alterman JF, Coles AH, Haraszti RA, Roux L, Godinho B, Echeverria D, Pears S, Iliopoulos J, Shanmugalingam R, Ogle R, Zsengeller ZK, Hennessy A, Karumanchi SA, Moore MJ, Khvorova A (2018) RNAi modulation of placental sFLT1 for the treatment of preeclampsia. Nat Biotechnol. https://doi.org/10.1038/nbt.4297
Chu C, Zhang QC, da Rocha ST, Flynn RA, Bharadwaj M, Calabrese JM, Magnuson T, Heard E, Chang HY (2015) Systematic discovery of Xist RNA binding proteins. Cell 161:404–416
Cao M, Zhao J, Hu G (2019) Genome-wide methods for investigating long noncoding RNAs. Biomed Pharmacother 111:395–401
Long Y, Wang X, Youmans DT, Cech TR (2017) How do lncRNAs regulate transcription? Sci Adv 3:eaao2110
Zhang Y, Liu T, Meyer CA, Eeckhoute J, Johnson DS, Bernstein BE, Nussbaum C, Myers RM, Brown M, Li W, Liu XS (2008) Model-based analysis of ChIP-Seq (MACS). Genome Biol 9:R137
Habener JF, Miller CP, Vallejo M (1995) cAMP-dependent regulation of gene transcription by cAMP response element-binding protein and cAMP response element modulator. Vitam Horm 51:1–57
Morishita K, Johnson DE, Williams LT (1995) A novel promoter for vascular endothelial growth factor receptor (flt-1) that confers endothelial-specific gene expression. J Biol Chem 270:27948–27953
Rutnam ZJ, Du WW, Yang W, Yang X, Yang BB (2014) The pseudogene TUSC2P promotes TUSC2 function by binding multiple microRNAs. Nat Commun 5:2914
Milligan MJ, Lipovich L (2014) Pseudogene-derived lncRNAs: emerging regulators of gene expression. Front Genet 5:476
Giulietti M, Righetti A, Principato G, Piva F (2018) LncRNA co-expression network analysis reveals novel biomarkers for pancreatic cancer. Carcinogenesis 39:1016–1025
Liu S, Wang Z, Chen D, Zhang B, Tian R, Wu J, Zhang Y, Xu K, Yang L, Cheng C, Ma J, Lv L, Zheng Y, Hu X, Zhang Y, Wang X, Li J (2017) Annotation and cluster analysis of spatiotemporal- and sex-related lncRNA expression in rhesus macaque brain. Genome Res. https://doi.org/10.1101/gr.217463.116
Leung DN, Smith SC, To K, Sahota DS, Baker PN (2001) Increased placental apoptosis in pregnancies complicated by preeclampsia. Am J Obstet Gynecol 184:1249–1250
Tomas SZ, Prusac IK, Roje D, Tadin I (2011) Trophoblast apoptosis in placentas from pregnancies complicated by preeclampsia. Gynecol Obstet Invest 71:250–255
Raguema N, Moustadraf S, Bertagnolli M (2020) Immune and apoptosis mechanisms regulating placental development and vascularization in preeclampsia. Front Physiol 11:98
Nakashima A, Cheng SB, Ikawa M, Yoshimori T, Huber WJ, Menon R, Huang Z, Fierce J, Padbury JF, Sadovsky Y, Saito S, Sharma S (2020) Evidence for lysosomal biogenesis proteome defect and impaired autophagy in preeclampsia. Autophagy 16:1771–1785
Engreitz JM, Pandya-Jones A, McDonel P, Shishkin A, Sirokman K, Surka C, Kadri S, Xing J, Goren A, Lander ES, Plath K, Guttman M (2013) The Xist lncRNA exploits three-dimensional genome architecture to spread across the X chromosome. Science 341:1237973
Quinn JJ, Ilik IA, Qu K, Georgiev P, Chu C, Akhtar A, Chang HY (2014) Revealing long noncoding RNA architecture and functions using domain-specific chromatin isolation by RNA purification. Nat Biotechnol 32:933–940
Simon MD, Wang CI, Kharchenko PV, West JA, Chapman BA, Alekseyenko AA, Borowsky ML, Kuroda MI, Kingston RE (2011) The genomic binding sites of a noncoding RNA. Proc Natl Acad Sci USA 108:20497–20502
Zou Y, Jiang Z, Yu X, Sun M, Zhang Y, Zuo Q, Zhou J, Yang N, Han P, Ge Z (2013) Upregulation of long noncoding RNA SPRY4-IT1 modulates proliferation, migration, apoptosis, and network formation in trophoblast cells HTR-8SV/neo. PLoS ONE 8:e79598
Zou YF, Sun LZ (2015) Long noncoding RNA HOTAIR modulates the function of trophoblast cells in pre-eclampsia. Sichuan Da Xue Xue Bao Yi Xue Ban 46(113–7):122
Chen H, Meng T, Liu X, Sun M, Tong C, Liu J, Wang H, Du J (2015) Long non-coding RNA MALAT-1 is downregulated in preeclampsia and regulates proliferation, apoptosis, migration and invasion of JEG-3 trophoblast cells. Int J Clin Exp Pathol 8:12718
Chen J, Xia J, Wang S, Wei Y, Wu J, Huang G, Chen F, Shi J (2012) Study on transcription regulation network in rheumatoid arthritis via bioinformatics analysis. Afr J Pharm Pharmacol 6:1435–1443
Fishilevich S, Nudel R, Rappaport N, Hadar R, Plaschkes I, Iny Stein T, Rosen N, Kohn A, Twik M, Safran M, Lancet D, Cohen D (2017) GeneHancer: genome-wide integration of enhancers and target genes in GeneCards. Database (Oxford). https://doi.org/10.1093/database/bax028
Tal R (2012) The role of hypoxia and hypoxia-inducible factor-1alpha in preeclampsia pathogenesis. Biol Reprod 87:134
Karakas D, Ozpolat B (2021) The role of LncRNAs in translation. Noncoding RNA 7:16
Huang J, Ling Z, Zhong H, Yin Y, Qian Y, Gao M, Ding H, Cheng Q, Jia R (2020) Distinct expression profiles of peptides in placentae from preeclampsia and normal pregnancies. Sci Rep 10:17558
Acknowledgements
We would like to thank all the participants and their families for their cooperation. We would also like to thank Yaqiang Xue and Chao Cheng from ABLife Inc. for their kind help in performing the experiments and bioinformatics analysis.
Funding
This work was supported by the National Natural Science Foundation of China (grant number 81760277), Natural Science Foundation of Jiangxi Province (20192BAB20502) and Science and Technology Project of Jiangxi Provincial Health Commission (20191773).
Author information
Authors and Affiliations
Contributions
XH and DC designed the project. SH and KX collected the PE samples. SX, and LZ performed ChIRP experiment. SX, YW, YX, DR, and BW performed data analysis. LZ and SX wrote the manuscript. All authors reviewed the manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors have no relevant financial or non-financial interests to disclose.
Ethical approval
Not applicable.
Consent to participate
Not applicable.
Consent to publish
Not applicable.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Zhao, L., Xin, S., Wu, Y. et al. Global DNA and protein interactomes of FLT1P1 (Fms-related tyrosine kinase 1 pseudogene 1) revealed its molecular regulatory functions associated with preeclampsia. Mol Biol Rep 50, 1267–1279 (2023). https://doi.org/10.1007/s11033-022-08070-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11033-022-08070-0