Publications by authors named "Huaxiang Zhao"

11 Publications

  • Page 1 of 1

Quantitative proteomic analysis of nonsyndromic orofacial cleft patient serum.

Oral Dis 2021 Jun 25. Epub 2021 Jun 25.

Key Laboratory of Shaanxi Province for Craniofacial Precision Medicine Research, College of Stomatology, Xi'an Jiaotong University, Xi'an, China.

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http://dx.doi.org/10.1111/odi.13947DOI Listing
June 2021

The cytokine FAM3B/PANDER is an FGFR ligand that promotes posterior development in .

Proc Natl Acad Sci U S A 2021 May;118(20)

Institute of Neuroscience, Translational Medicine Institute, Health Science Center, Xi'an Jiaotong University, 710061 Xi'an, China;

Fibroblast growth factor (FGF)/extracellular signal-regulated kinase (ERK) signaling plays a crucial role in anterior-posterior (A-P) axial patterning of vertebrate embryos by promoting posterior development. In our screens for novel developmental regulators in embryos, we identified Fam3b as a secreted factor regulated in ectodermal explants. Family with sequence similarity 3 member B (FAM3B)/PANDER (pancreatic-derived factor) is a cytokine involved in glucose metabolism, type 2 diabetes, and cancer in mammals. However, the molecular mechanism of FAM3B action in these processes remains poorly understood, largely because its receptor is still unidentified. Here we uncover an unexpected role of FAM3B acting as a FGF receptor (FGFR) ligand in embryos. messenger RNA (mRNA) is initially expressed maternally and uniformly in the early embryo and then in the epidermis at neurula stages. Overexpression of mRNA inhibited cephalic structures and induced ectopic tail-like structures. Recombinant human FAM3B protein was purified readily from transfected tissue culture cells and, when injected into the blastocoele cavity, also caused outgrowth of tail-like structures at the expense of anterior structures, indicating FGF-like activity. Depletion of by specific antisense morpholino oligonucleotides in resulted in macrocephaly in tailbud tadpoles, rescuable by FAM3B protein. Mechanistically, FAM3B protein bound to FGFR and activated the downstream ERK signaling in an FGFR-dependent manner. In embryos, FGFR activity was required epistatically downstream of Fam3b to mediate its promotion of posterior cell fates. Our findings define a FAM3B/FGFR/ERK-signaling pathway that is required for axial patterning in embryos and may provide molecular insights into FAM3B-associated human diseases.
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http://dx.doi.org/10.1073/pnas.2100342118DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8158011PMC
May 2021

A novel mutation revealed the cause of cleft lip and/or palate in a Chinese family.

Genes Dis 2020 Sep 18;7(3):440-447. Epub 2019 Dec 18.

Central Laboratory, Peking University School and Hospital of Stomatology, Beijing, China.

Cleft lip and/or palate (CL/P) is a most common craniofacial birth defect which has multifactorial etiology. In our study, we aimed to discover the underlying etiological gene variation in a Chinese family diagnosed as non-syndromic CL/P (NSCL/P). The blood sample of the proband and her parents were detected by whole exome sequencing. The Mendelian inheritance pattern, allele frequency, variation location, function analysis and literature search were applied to filtrate and screen the mutation. Besides, the candidates were confirmed by Sanger sequencing. We meanwhile explored the conservative analysis and protein homology simulation. As a result, a start-lost mutation c.1A > GAtg/Gtg in the Frizzled-6 () gene predicting p.Met1 was detected. The variation has not been reported before and was predicted to be harmful. The alteration caused missing of two starting amino acids that are evolutionarily conserved for FZD6 protein. Moreover, the specific structure of the mutant protein obviously changed according to the results of the homologous model. In conclusion, the results suggest c.1A > GAtg/Gtg in the (NM_001164616) might be the genetic etiology for non-syndromic CL/P in this pedigree. Furthermore, this finding provided new etiologic information, supplementing the evidence that is a strong potential gene for CL/P.
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http://dx.doi.org/10.1016/j.gendis.2019.12.003DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7452514PMC
September 2020

Functional Characterization of a Novel IRF6 Frameshift Mutation From a Van Der Woude Syndrome Family.

Front Genet 2020 4;11:562. Epub 2020 Jun 4.

Central Laboratory, Peking University School and Hospital of Stomatology, Beijing, China.

Background: Loss-of-function mutations in interferon regulatory factor-6 () are responsible for about 70% of cases of Van Der Woude Syndrome (VWS), an autosomal dominant developmental disorder characterized by pits and/or sinuses of the lower lip and cleft lip, cleft palate, or both.

Methods: We collected a Chinese Han VWS pedigree, performed sequencing and screening for the causal gene mutant. Initially, species conservation analysis and homology protein modeling were used to predict the potential pathogenicity of mutations. To test whether a VWS family-derived mutant variant of retained function, we carried out rescue assays in maternal-null mutant zebrafish embryos. To assess protein stability, we overexpressed reference and family-variants of IRF6 .

Results: We focused on a VWS family that includes a son with bilateral lip pits, uvula fissa and his father with bilateral cleft lip and palate. After sequencing and screening, a frameshift mutation of was identified as the potential causal variant (NM.006147.3, c.1088-1091delTCTA; p.Ile363ArgfsTer33). The residues in this position are strongly conserved among species and homology modeling suggests the variant alters the protein structure. In maternal-null mutant zebrafish embryos the periderm differentiates abnormally and the embryos rupture and die during gastrulation. Injection of mRNA encoding the reference variant of human IRF6, but not of the frame-shift variant, rescued such embryos through gastrulation. Upon overexpression in HEK293FT cells, the IRF6 frame-shift mutant was relatively unstable and was preferentially targeted to the proteasome in comparison to the reference variant.

Conclusion: In this VWS pedigree, a novel frameshift of was identified as the likely causative gene variant. It is a lost function mutation which could not rescue abnormal periderm phenotype in maternal-null zebrafish and which causes the protein be unstable through proteasome-dependent degradation.
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http://dx.doi.org/10.3389/fgene.2020.00562DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7289175PMC
June 2020

Identification of rare nonsense variant causing orofacial cleft in a Chinese family and an up-to-date genotype-phenotype analysis.

Genes Dis 2021 Sep 8;8(5):689-697. Epub 2020 Jan 8.

Department of Orthodontics, Peking University School and Hospital of Stomatology, Beijing, 100081, PR China.

The Patched 1 () gene encodes a membrane receptor involved in the Hedgehog (Hh) signaling pathway, an abnormal state of which may result in congenital defects or human tumors. In this study, we conducted whole-exome sequencing on a three-generation Chinese family characterized with variable penetrance of orofacial clefts. A rare heterozygous variant in the gene (c.2833C > T p.R945X) was identified as a disease-associated mutation. Structural modeling revealed a truncation starting from the middle of the second extracellular domain of PTCH1 protein. This may damage its ligand recognition and sterol transportation abilities, thereby affecting the Hh signaling pathway. Biochemical assays indicated that the R945X protein had reduced stability compared to the wild-type . In addition, we reviewed the locations and mutation types of variants in individuals with clefting phenotypes, and analyzed the associations between clefts and locations or types of variants within . Our findings provide further evidence that variants result in orofacial clefts, and contributed to genetic counseling and clinical surveillance in this family.
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http://dx.doi.org/10.1016/j.gendis.2019.12.010DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8278535PMC
September 2021

Three GLI2 mutations combined potentially underlie non-syndromic cleft lip with or without cleft palate in a Chinese pedigree.

Mol Genet Genomic Med 2019 09 6;7(9):e714. Epub 2019 Aug 6.

Department of Orthodontics, Peking University School and Hospital of Stomatology, Beijing, PR China.

Background: Nonsyndromic cleft lip with or without cleft palate (NSCL/P) is the most common craniofacial birth defect. Its etiology is complex and it has a lifelong influence on affected individuals. Despite many studies, the pathogenic gene alleles are not completely clear. Here, we recruited a Chinese NSCL/P family and explored the candidate causative variants in this pedigree.

Methods: We performed whole-exome sequencing on two patients and two unaffected subjects of this family. Variants were screened based on bioinformatics analysis to identify the potential etiological alleles. Species conservation analysis, mutation function prediction, and homology protein modeling were also performed to preliminarily evaluate the influence of the mutations.

Results: We identified three rare mutations that are located on a single chromatid (c.2684C > T_p.Ala895Val, c.4350G > T_p.Gln1450His, and c.4622C > A_p.Ser1541Tyr) in GLI2 as candidate causative variants. All of these three mutations were predicted to be deleterious, and they affect amino acids that are conserved in many species. The mutation c.2684C > T was predicted to affect the structure of the GLI2 protein.

Conclusion: Our results further demonstrate that GLI2 variants play a role in the pathogenesis of NSCL/P, and the three rare missense mutations combined are probably the potential disease-causing variants in this family.
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http://dx.doi.org/10.1002/mgg3.714DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6732289PMC
September 2019

A novel IRF6 mutation causing non-syndromic cleft lip with or without cleft palate in a pedigree.

Mutagenesis 2018 09;33(3):195-202

Central Laboratory, Peking University School and Hospital of Stomatology, Beijing, PR China.

Non-syndromic cleft lip with or without cleft palate (NSCLP) is the most common congenital craniofacial malformation, and its harmful influence on affected individuals is apparent. Despite many studies, the causative genes and their mechanisms are not completely clear. We recruited a Han Chinese NSCLP family and explored the causative variant in this pedigree. We performed whole-exome sequencing on two patients. Bioinformatics screening and analysis were used to identify the mutation. We also performed species conservation analysis, mutation function predictions, and homology protein modelling to evaluate the influence of the mutation. We identified a rare mutation in interferon regulatory factor 6 (IRF6) (c.26G>A; p.Arg9Gln) as a candidate of causative mutation. This mutation was predicted to be deleterious. The codon is conserved in many species. The residue change caused by this mutation would affect the structure of IRF6 to a degree. Our study suggested that the rare IRF6 variant is probably the pathogenic mutation in this family. Our result adds evidence that IRF6 variants play a role in the aetiology of orofacial clefts.
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http://dx.doi.org/10.1093/mutage/gey012DOI Listing
September 2018

A novel PTCH1 mutation underlies nonsyndromic cleft lip and/or palate in a Han Chinese family.

Oral Dis 2018 Oct 9;24(7):1318-1325. Epub 2018 Jul 9.

Central Laboratory, Peking University School and Hospital of Stomatology, Beijing, China.

Objectives: Cleft lip and/or palate (CL/P) is the most common craniofacial congenital disease, and it has a complex aetiology. This study aimed to identify the causative gene mutation of a Han Chinese family with CL/P.

Subjects And Methods: Whole exome sequencing was conducted on the proband and her mother, who exhibited the same phenotype. A Mendelian dominant inheritance model, allele frequency, mutation regions, functional prediction and literature review were used to screen and filter the variants. The candidate was validated by Sanger sequencing. Conservation analysis and homology modelling were conducted.

Results: A heterozygous missense mutation c.1175C>T in the PTCH1 gene predicting p.Ala392Val was identified. This variant has not been reported and was predicted to be deleterious. Sanger sequencing verified the variant and the dominant inheritance model in the family. The missense alteration affects an amino acid that is evolutionarily conserved in the first extracellular loop of the PTCH1 protein. The local structure of the mutant protein was significantly altered according to homology modelling.

Conclusions: Our findings suggest that c.1175C>T in PTCH1 (NM_000264) may be the causative mutation of this pedigree. Our results add to the evidence that PTCH1 variants play a role in the pathogenesis of orofacial clefts.
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http://dx.doi.org/10.1111/odi.12915DOI Listing
October 2018

Nell-1-ΔE, a novel transcript of Nell-1, inhibits cell migration by interacting with enolase-1.

J Cell Biochem 2018 07 16;119(7):5725-5733. Epub 2018 Apr 16.

Central Laboratory, Peking University School and Hospital of Stomatology, Beijing, P. R. China.

NELL-1 is a secreted protein that was originally found to be upregulated in pathologically fusing and fused sutures in non-syndromic unilateral coronal synostosis patients. Apart from the ability of NELL-1 to promote osteogenesis in long and craniofacial bones, NELL-1 reportedly inhibits the formation of several benign and malignant tumors. We previously identified a novel transcript of Nell-1 that lacked a calcium-binding epidermal growth factor (EGF)-like domain compared with full-length Nell-1; this new transcript was named Nell-1-ΔE. Three obvious structural differences between these two isoforms were revealed by homology modeling. Furthermore, the recombinant Nell-1-ΔE protein, but not the full-length Nell-1 protein, inhibited cell migration in vitro. However, full-length Nell-1 and Nell-1-ΔE proteins were present in similar subcellular locations and displayed similar expression patterns in both the intracellular and extracellular spaces. The results from the co-immunoprecipitation and liquid chromatography/tandem mass spectrometry analyses using two cell lines demonstrated that Nell-1-ΔE but not full-length Nell-1 interacted with enolase-1 in the extracellular spaces of both cell lines. The results of wound healing assays using ENO-1-overexpressing cells treated with full-length Nell-1/Nell-1-ΔE suggested that Nell-1-ΔE inhibited cell migration by interacting with ENO-1. Our study indicated that the novel transcript Nell-1-ΔE, but not full-length Nell-1, might be a candidate tumor suppressor factor for basic research and clinical practice.
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http://dx.doi.org/10.1002/jcb.26756DOI Listing
July 2018

Is MTHFD1 polymorphism rs 2236225 (c.1958G>A) associated with the susceptibility of NSCL/P? A systematic review and meta-analysis.

F1000Res 2015 4;4:142. Epub 2015 Jun 4.

Department of Orthodontics, Peking University School and Hospital of Stomatology, Peking, 100081, China.

Aims: To investigate the association between the methylenetetrahydrofolate dehydrogenase 1 (MTHFD1) polymorphism rs 2236225 (c.1958G>A) and susceptibility to non-syndromic cleft of the lip and/or palate (NSCL/P).

Methods: An extensive literature review has been conducted using PubMed, Web of Science, Cochrane Library, Google Scholar, the China National Knowledge Infrastructure (CNKI), and Wanfang Database for eligible researches. The terms for searching were "cleft lip OR cleft palate OR CLP OR CL/P OR oral facial cleft OR OFC" AND "methylenetetrahydrofolate dehydrogenase (NADP+ dependent) 1 OR methenyltetrahydrofolate cyclohydrolase formyltetrahydrofolate synthetase OR MTHFD1 OR MTHFD". Two independent researchers screened, evaluated and extracted the data of included studies. The pooled odds ratios (OR) with 95% confidence intervals (95% CI) were calculated by random effects model under five gene models. Subgroup, sensitivity analysis and publication bias were also assessed.

Results: Ten case-control studies have been included in the systematic review and eight studies have been considered for the meta-analysis. Overall, the MTHFD1 polymorphism rs2236225 and the risk of NSCL/P showed pooled OR (95% CI) of 1.02 (0.86-1.21) under allelic model. A higher degree of heterogeneity was observed in Asian countries (I (2) = 75.6%) compared to non-Asian countries (I (2) = 48.9%). Similar consequence appeared in the subgroup of children (I (2) = 78.6%) compared with that of mothers (I (2) = 0.0%). There was no significant difference in the publication bias by the Begg's funnel plot (P = 0.711) and Egger's regression test (P = 0.746).

Conclusion: Our assessment suggested there was no significant association between the MTHFD1 polymorphism rs 2236225 (c.1958G>A) and the susceptibility to NSCL/P. Further investigations using a large sample size and a more advanced technique should be adopted to reach a more precise conclusion in the future.
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http://dx.doi.org/10.12688/f1000research.6425.2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4722688PMC
February 2016

Mutation identification of the DSPP in a Chinese family with DGI-II and an up-to-date bioinformatic analysis.

Genomics 2012 Apr 31;99(4):220-6. Epub 2012 Jan 31.

Department of Genetics and Molecular Biology, Xi'an Jiaotong University School of Medicine, Xi'an, Shaanxi 710061, PR China.

In this study, through linkage analysis of a four-generation Chinese family with multiple members afflicted with DGI (type II), we identified a novel missense mutation in DSPP. The mutation was located in exon 2 at the second nucleotide position of the last codon and resulted in a substitution of a proline with a leucine residue (c.50C>T, p.P17L, g.50C>T). To assess the potential effects of this novel mutation, we utilized various bioinformatics analysis programs. The results indicate that the mutation likely affects protein cleavage/trafficking. We also analyzed previously reported mutations of DSPP. In summary, our finding supports that the genomic sequence that corresponds to the P17 residue of DSPP is a mutational hotspot and P17 may be critical for the function of DSPP.
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http://dx.doi.org/10.1016/j.ygeno.2012.01.006DOI Listing
April 2012
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