Progress of epigenetic modification of SATB2 gene in the pathogenesis of non-syndromic cleft lip and palate.

Non-syndromic Cleft Lip and Palate (NSCLP) is one of the most common congenital craniofacial malformations. However, there is no enough knowledge about its mechanism, even through many relevant studies verify that cleft lip and palate is caused by interactions between environmental and genetic factors. SATB2 gene is one of the most common candidate genes of NSCLP, and the development of epigenetics provides a new direction on pathogenesis of cleft lip and palate. This review summarizes SATB2 gene in the pathogenesis of non-syndromic cleft lip and palate, expecting to provide strategies to prevent and treat cleft and palate in the future.

Reading-related Brain Function Restored to Normal After Articulation Training in Patients with Cleft Lip and Palate: An fMRI Study.

Cleft lip and/or palate (CLP) are the most common craniofacial malformations in humans. Speech problems often persist even after cleft repair, such that follow-up articulation training is usually required. However, the neural mechanism behind effective articulation training remains largely unknown. We used fMRI to investigate the differences in brain activation, functional connectivity, and effective connectivity across CLP patients with and without articulation training and matched normal participants. We found that training promoted task-related brain activation among the articulation-related brain networks, as well as the global attributes and nodal efficiency in the functional-connectivity-based graph of the network. Our results reveal the neural correlates of effective articulation training in CLP patients, and this could contribute to the future improvement of the post-repair articulation training program.

Association of rs2013162 and rs2235375 Polymorphisms in IRF6 Gene with Susceptibility to Non-Syndromic Cleft Lip and Palate.

Background: Non-syndromic cleft lip occurs by the interaction of environmental and genetic factors. The purpose of the current study was to analyze the association of Single Nucleotide Polymorphisms (SNPs) in IRF6 and NSCL/P in an Iranian population. Methods: A group of 105 children with NSCL/P and 185 normal controls were included in the current study. Genotyping of IRF6 rs2013162 and rs2235375 was performed by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) method. Results: A substantial association of AA and CA genotypes in rs2013162 with the risk of NSCL/P (AA vs. CC; OR=2.36; 95%CI [1.05-5.29], p=0.004; and CA vs. CC; OR=0.47; 95%CI [0.28-0.79], p=0.018) was found. However, there were no important associations between A allele and risk of NSCL/P (p=0.980). According to logistic regression analysis results, subjects with GG genotype and G allele in rs2235375 polymorphism had increased risk of NSCL/P. Conclusion: The IRF6 polymorphisms are associated with the susceptibility to NSCL/P in Iranian population.

Random Network and Non-rich-club Organization Tendency in Children With Non-syndromic Cleft Lip and Palate After Articulation Rehabilitation: A Diffusion Study.

OBJECTIVE: The neuroimaging pattern in brain networks after articulation rehabilitation can be detected using graph theory and multivariate pattern analysis (MVPA). In this study, we hypothesized that the characteristics of the topology pattern of brain structural network in articulation-rehabilitated children with non-syndromic cleft lip and palate (NSCLP) were similar to that in healthy comparisons. METHODS: A total of 28 children with NSCLP and 28 controls with typical development were scanned for diffusion tensor imaging on a 3T MRI scanner. Structural networks were constructed, and their topological properties were obtained. Besides, the Chinese language clear degree scale (CLCDS) scores were used for correlation analysis with topological features in patients with NSCLP. RESULTS: The NSCLP group showed a similar rich-club connection pattern, but decreased small-world index, normalized rich-club coefficient, and increased connectivity strength of connections compared to controls. The univariate and multivariate patterns of the structural network in articulation-rehabilitated children were primarily in the feeder and local connections, covering sensorimotor, visual, frontoparietal, default mode, salience, and language networks, and orbitofrontal cortex. In addition, the connections that were significantly correlated with the CLCDS scores, as well as the weighted regions for classification, were chiefly distributed in the dorsal and ventral stream associated with the language networks of the non-dominant hemisphere. CONCLUSION: The average level rich-club connection pattern and the compensatory of the feeder and local connections mainly covering language networks may be related to the CLCDS in articulation-rehabilitated children with NSCLP. However, the patterns of small-world and rich-club structural organization in the articulation-rehabilitated children exhibited a random network and non-rich-club organization tendency. These findings enhanced the understanding of neuroimaging patterns in children with NSCLP after articulation rehabilitation.

Dental Characteristics of Different Types of Cleft and Non-cleft Individuals.

OBJECTIVE: The objective of this study was to compare the novel artificial intelligence (A.I.)-driven lateral cephalometric (Late. Ceph.) analysis of 14 different dental characteristics (DC) among different types of cleft lip and palate (CLP) and non-cleft (NC) individuals. MATERIALS AND METHODS: A retrospective study was conducted on 123 individuals [31 = NC, 29 = BCLP (bilateral cleft lip and palate), 41 = UCLP (unilateral cleft lip and palate), 9 = UCLA (unilateral cleft lip and alveolus), and 13 = UCL (unilateral cleft lip)] with an average age of 14.77 years. Demographic details were gathered from the clinical records. A novel artificial intelligence-driven Webceph software has been used for the Late. Ceph. analysis. A total of 14 different types of angular and linear DC measurements were analyzed and compared among groups. Two-way ANOVA and multiple-comparison statistics tests were applied to see the differences between gender and among different types of CLP versus NC subjects. RESULTS: Of the 14 DC tested, no significant gender disparities were found (p > 0.05). In relation to different types of CLP versus NC subjects, 8 over 14 DC were statistically significant (p < 001 to p = 0.03). Six other DC variables show insignificant (p > 0.05) noteworthy alterations in relation to type of CLP. CONCLUSION: Based on the results, type of CLP revealed significantly altered DC compared to NC. Among different types of CLP, BCLP exhibited a maximum alteration in different DC.

Genetic variation of IRF6 and TGFA genes in an HIV-exposed newborn with non-syndromic cleft lip palate.

We present here the first reported case of a non-syndromic cleft lip and palate (NSCLP) in an HIV-exposed newborn of a mother on antiretroviral therapy (ART) in Indonesia. Genetic testing was performed to confirm a suspected genetic condition. Genomic DNA was extracted from the blood, and genetic variations of the interferon regulatory factor 6 (IRF6) rs642961 (Mspl) (G>A) and transforming growth factor alpha (TGFA) BamHI (rs11466297, A>C) and RsaI (rs3732248, C>T) were performed by PCR-RFLP and IRF6 gene analysis by PCR sequencing. Genotyping of DNA sequence variants in the IRF6 gene showed both parents had genotype GA, while the child had genotype GG (genotype wild type). There was no difference observed in the TGFA BamHI gene variant between the child and her mother and father that were wild-type polymorphisms (normal), while the Rsa1 polymorphisms of them were heterozygotes. A genetic variant of IRF6 might be a protective factor for NSCLP, while Rsa1 gene variant (A) allele can be considered to be the risk factor associated with NSCLP development. This case report also highlights the possible etiologic role of ART in NSCLP; therefore, early control of adverse effects of ART might be an important factor in decreasing the incidence of the congenital anomalies in HIV-infected children.

Articulation rehabilitation induces cortical plasticity in adults with non-syndromic cleft lip and palate.

In this study, we investigated brain morphological changes in adults with non-syndromic cleft lip and palate (NSCLP) after articulation rehabilitation (AR). High-resolution T1 weighted brain magnetic resonance imaging data were analyzed from 45 adults with NSCLP after palatoplasty: 24 subjects were assessed before AR (bNSCLP) and 21 subjects were assessed after AR (aNSCLP). In addition, there were 24 age and sex matched controls. Intergroup differences of grey matter volume were evaluated as a comprehensive measure of the cortex; cortical thickness and cortical complexity (gyrification and fractal dimensions) were also analyzed. As compared to controls, the bNSCLP subjects exhibited altered indexes in frontal, temporal, and parietal lobes; these morphological changes are characteristic for adults with NSCLP. Importantly, as compared to the bNSCLP and control subjects, the aNSCLP subjects exhibited cortical plasticity in the regions involved in language, auditory, pronunciation planning, and execution functions. The AR-mediated cortical plasticity in aNSCLP subjects may be caused by AR-induced cortical neurogenesis, which might reflect the underlying neural mechanism during AR.

Overexpression of YOD1 Promotes the Migration of Human Oral Keratinocytes by Enhancing TGF-ß3 Signaling.

OBJECTIVE: To investigate the effects of YOD1 overexpression on the proliferation and migration of human oral keratinocytes (HOKs), and to clarify whether the mechanisms involve transforming growth factor-ß (TGF-ß) signaling. METHODS: HOKs were transfected with the plasmid pEGFP-N3-YOD1 containing YOD1. The mRNA levels of YOD1 and TGF-ß were determined by qPCR. The protein expressions of YOD1, TGF-ß, Smad2/3, Smad4, and phospho-Smad2/3 were determined by western blotting. Cell proliferation and migration were evaluated by Cell Counting Kit-8 assay and wound healing assay, respectively. RESULTS: The mRNA and protein levels of YOD1 were higher in HOKs transfected with YOD1. YOD1 overexpression significantly enhanced the migration of HOKs. The mRNA and protein levels of TGF-ß3 were increased by YOD1 overexpression. HOKs transfected with YOD1 exhibited increased phospho-Smad2/3 levels. CONCLUSION: YOD1 overexpression enhances cell migration by promoting TGF-ß3 signaling which may play an important role in lip and palate formation. YOD1 mutation may contribute to aberrant TGF-ß3 signaling associated with decreased cell migration resulting in NSCLP.

Overexpression of YOD1 Promotes the Migration of Human Oral Keratinocytes by Enhancing TGF-β3 Signaling / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate the effects of YOD1 overexpression on the proliferation and migration of human oral keratinocytes (HOKs), and to clarify whether the mechanisms involve transforming growth factor-β (TGF-β) signaling.</p><p><b>METHODS</b>HOKs were transfected with the plasmid pEGFP-N3-YOD1 containing YOD1. The mRNA levels of YOD1 and TGF-β were determined by qPCR. The protein expressions of YOD1, TGF-β, Smad2/3, Smad4, and phospho-Smad2/3 were determined by western blotting. Cell proliferation and migration were evaluated by Cell Counting Kit-8 assay and wound healing assay, respectively.</p><p><b>RESULTS</b>The mRNA and protein levels of YOD1 were higher in HOKs transfected with YOD1. YOD1 overexpression significantly enhanced the migration of HOKs. The mRNA and protein levels of TGF-β3 were increased by YOD1 overexpression. HOKs transfected with YOD1 exhibited increased phospho-Smad2/3 levels.</p><p><b>CONCLUSION</b>YOD1 overexpression enhances cell migration by promoting TGF-β3 signaling which may play an important role in lip and palate formation. YOD1 mutation may contribute to aberrant TGF-β3 signaling associated with decreased cell migration resulting in NSCLP.</p>

[Cleft lip and palate]. / Lippen-Kiefer-Gaumen-Spalten.

BACKGROUND: Cleft lip and palate (CLP) represents a group of malformations of unknown etiology but similar phenotypes. This implies consequences for the diagnostics, therapy, prevention, prognosis and risk estimation. OBJECTIVE: Definition of CLP subtypes and the embryonic development, clarification of correlations and differences between entities using epidemiological data, overview of the present state of genetic analyses, correlation to syndromes, sequences and associations and resulting consequences for clinical practice. MATERIAL AND METHODS: Update on embryological development of the face, summary of epidemiological and genetic studies and considerations on pedopathological and forensic aspects. RESULTS: Syndromic and non-syndromic CLP exhibit different and highly variable etiologies, therapeutic needs and prognosis. A thorough understanding is mandatory to distinguish between the different subgroups. In addition to specific aspects of CLP for the pediatric (forensic) pathologist this article provides an overall view of the topic which aims to help understand these malformations.

Single nucleotide polymorphism of bone morphogenetic protein 4 gene: A risk factor of non-syndromic cleft lip with or without palate.

BACKGROUND: The bone morphogenetic protein (BMP) signalling pathway is crucial in a number of developmental processes and is critical in the formation of variety of craniofacial elements including cranial neural crest, facial primordium, tooth, lip and palate. It is an important mediator in regulation of lip and palate fusion, cartilage and bone formation. AIM: To study the role of mutation of BMP4 genes in the aetiology of non-syndromic cleft lip with or without palate (NSCL ± P) and identify it directly from human analyses. MATERIALS AND METHODS: A case-control study was done to evaluate whether BMP4T538C polymorphism, resulting in an amino acid change of Val=Ala (V152A) in the polypeptide, is associated with NSCL ± P in an Indian paediatric population. Genotypes of 100 patients with NSCL ± P and 100 controls (in whom absence of CL ± P was confirmed in three generations) were detected using a polymerase chain reaction-restriction fragment length polymorphism strategy. Logistic regression was performed to evaluate allele and genotype association with NSCLP. RESULTS: Results showed significant association between homozygous CC genotype with CL ± P (odds ratio [OR]-5.59 and 95% confidence interval [CI] = 2.85-10.99). The 538C allele carriers showed an increased risk of NSCL ± P as compared with 538 T allele (OR - 4.2% CI = 2.75-6.41). CONCLUSION: This study suggests an association between SNP of BMP4 gene among carriers of the C allele and increased risk for NSCLP in an Indian Population. Further studies on this aspect can scale large heights in preventive strategies for NSCLP that may soon become a reality.

Association between IRF6 gene SNPs and non-syndromic cleft lip with or without cleft palate in Xinjiang Uyghur population / 实用口腔医学杂志

Objective:To study the association between rs2235371,rs2013162,rs2235377 SNPs in interferon regulatory factor 6 (IRF6)gene and non-syndromic cleft lip with or without cleft palate(NSCL/P)in Xinjiang Uyghur population.Methods:100 Uyghur NSCL/P patients from Xinjiang were included in the case group and 60 Uyghur inpatients with upper respiratory tract infection were se-lected in the control group.Next,generation sequencing was used,DNA sequencing results were compared with the information on the genome database and genetic analysis were made.Results:There were no significant differences in the frequency distribution of both genotypes and alles when the cases were campared with the controls at the rs2235371,rs2013162 and rs2235377 loci(P >0.05). Above three loci were located in the same block,rs2235371 and rs2235377 loci presents the strong linkage disequilibrium(r2 =0.949, D'=0.974).Possible haplotypes were:CCT >CAT >TAC,and there was no significant difference between the cases and controls in haplotype distribution(P >0.05).Conclusion:Polymorphisms of rs2235371,rs2013162 and rs2235377 in IRF6 gene may be associ-ated with NSCL/P in Xinjiang Uygur people.

Both LCCL-domains of human CRISPLD2 have high affinity for lipid A.

The LCCL-domain is a recently defined protein module present in diverse extracellular multidomain proteins. Practically nothing is known about the molecular function of these domains; based on functional features of proteins harboring LCCL-domains it has been suggested that these domains might function as lipopolysaccharide-binding domains. Here we show that the two LCCL-domains of human CRISPLD2 protein, a lipopolysaccharide-binding serum protein involved in defense against endotoxin shock, have higher affinity for the lipid A, the toxic moiety of lipopolysaccharides than for ipopolysaccharide. Our observation that the LCCL-domains of CRISPLD2 are specific for the toxic lipid A moiety of the endotoxin suggests that it may block the interaction between endotoxins and the host endotoxin receptors without interfering with the development of antibacterial immunity against the polysaccharide moiety of LPS. We suggest that the anti-inflammatory function of CRISPLD2 protein may account for its role in various pathological and developmental processes.