Functional Analysis of IFT172 Mutations in Families with Non-syndromic Cleft Lip with or without Palate

doi:10.13701/j.cnki.kqyxyj.2025.06.011

Abstract

Abstract: Objective: To further define the pathogenic mechanism of intraflagellar transport 172 (IFT172) in cleft lip and palate through functional analysis of IFT172 mutations identified in two families with non-syndromic cleft lip with or without palate. Methods: The IFT172 mutations identified in the previous work were subject to conservation analysis and protein structure prediction. Wild-type and mutant plasmids were constructed and transfected into human embryonic palatal mesenchyme (HEPM) cells. Transcriptome sequencing was performed and differentially expressed genes were analysed. Results: Two missense mutations in the IFT172, c.4163A>G (p.Y1388C) and c.1507A>G (p.R503G), were highly conservative among different species, and both mutations resulted in altered spatial conformation of the protein. The differentially expressed genes were enriched in pathways such as necroptosis and regulation of cell pluripotency. Conclusion: The missense mutations in the IFT172 may be involved in cleft lip and palate by altering protein conformation and affecting necroptosis and cell pluripotency.

Key words: IFT172, primary cilia, non-syndromic cleft lip with or without palate, transcriptome sequencing

LUO Wendi, CAO Haiyan, ZUO Yining, HE Miao. Functional Analysis of IFT172 Mutations in Families with Non-syndromic Cleft Lip with or without Palate[J]. Journal of Oral Science Research, 2025, 41(6): 517-520.

References

[1] Xu DP, Qu WD, Sun C, et al. A study on environmental factors for nonsyndromic cleft lip and/or palate[J]. J Craniofac Surg, 2018, 29(2): 364-367.
[2] Roth DM, Bayona F, Baddam P, et al. Craniofacial development: Neural crest in molecular embryology [J]. Head Neck Pathol, 2021, 15(1): 1-15.
[3] Schock EN, Brugmann SA. Discovery, diagnosis, and etiology of craniofacial ciliopathies [J]. Cold Spring Harb Perspect Biol, 2017, 9(9): a028258.
[4] Ma R, Kutchy NA, Chen L, et al. Primary cilia and ciliary signaling pathways in aging and age-related brain disorders [J]. Neurobiol Dis, 2022, 163: 105607.
[5] Pakvasa M, Haravu P, Boachie-Mensah M, et al. Notch signaling: Its essential roles in bone and craniofacial development [J]. Genes Dis, 2020, 8(1): 8-24.
[6] Barba A, Urbina C, Maili L, et al. Association of IFT88 gene variants with nonsyndromic cleft lip with or without cleft palate [J]. Birth Defects Res, 2019, 111(11): 659-665.
[7] Tian H, Feng J, Li J, et al. Intraflagellar transport 88 (IFT88) is crucial for craniofacial development in mice and is a candidate gene for human cleft lip and palate [J]. Hum Mol Genet, 2017, 26(5): 860-872.
[8] Zheng NX, Miao YT, Zhang X, et al. Primary cilia-associated protein IFT172 in ciliopathies [J]. Front Cell Dev Biol, 2023, 11: 1074880.
[9] Bujakowska KM, Zhang Q, Siemiatkowska AM, et al. Mutations in IFT172 cause isolated retinal degeneration and Bardet-Biedl syndrome [J]. Hum Mol Genet, 2015, 24(1): 230-242.
[10] Pruski M, Hu L, Yang C, et al. Roles for IFT172 and primary cilia in cell migration, cell division, and neocortex development [J]. ront Cell Dev Biol, 2019, 7: 287.
[11] Askarian S, Gholami M, Khalili-Tanha G, et al. The genetic factors contributing to the risk of cleft lip-cleft palate and their clinical utility [J]. Oral Maxillofac Surg, 2023, 27(2): 177-186.
[12] Wattanawong K, Rattanasiri S, McEvoy M, et al. Association between IRF6 and 8q24 polymorphisms and nonsyndromic cleft lip with or without cleft palate: Systematic review and meta-analysis [J]. Birth Defects Res A Clin Mol Teratol, 2016, 106(9): 773-788.
[13] de Araujo TK, Secolin R, Félix TM, et al. A multicentric association study between 39 genes and nonsyndromic cleft lip and palate in a Brazilian population [J]. J Craniomaxillofac Surg, 2016, 44(1): 16-20.
[14] Yan C, Deng-Qi H, Li-Ya C, et al. Transforming growth factor alpha taq Ⅰ polymorphisms and nonsyndromic cleft lip and/or palate risk: A meta-analysis [J]. Cleft Palate Craniofac J, 2018, 55(6): 814-820.
[15] Shi M, Mostowska A, Jugessur A, et al. Identification of microdeletions in candidate genes for cleft lip and/or palate [J]. Birth Defects Res A Clin Mol Teratol, 2009, 85(1): 42-51.
[16] Ingham PW, McMahon AP. Hedgehog signaling in animal development: paradigms and principles [J]. Genes Dev, 2001, 15(23): 3059-3087.
[17] Friedland-Little JM, Hoffmann AD, Ocbina PJR, et al. A novel murine allele of Intraflagellar Transport Protein 172 causes a syndrome including VACTERL-like features with hydrocephalus [J]. Hum Mol Genet, 2011, 20(19): 3725-3737.
[18] Ocbina PJ, Tuson M, Anderson K. Primary cilia are not required for normal canonical Wnt signaling in the mouse embryo [J]. PLoS One, 2009, 4(8):e6839.
[19] Corbit KC, Shyer AE, Dowdle WE, et al. Kif3a constrains β-catenin-dependent Wnt signalling through dual ciliary and non-ciliary mechanisms [J]. Nat Cell Biol, 2008, 10(1): 70-76.
[20] Buschbeck M, Hake SB. Variants of core histones and their roles in cell fate decisions, development and cancer [J]. Nat Rev Mol Cell Biol, 2017, 18(5): 299-314.
[21] Shi L, Liang Y, Yang L, et al. All-trans retinoic acid regulates miR-106a-5p inhibition of autophagic in developing cleft palates [EB]. bioRxiv, 2020.