Transcriptome analyses of murine right and left maxilla-mandibular complex.

OBJECTIVE The objective of the study was to investigate differential gene expression between murine right and left maxilla-mandibular (MxMn) complexes. SETTING AND SAMPLE POPULATION Wild-type (WT) C57BL/6 embryonic (E) day 14.5 (n = 3) and 18.5 (n = 3) murine embryos. METHODS The E14.5 and 18.5 embryos were harvested and hemi-sectioned the MxMn complexes into right and left halves in the mid-sagittal plane. We isolated total RNA using Trizol reagent and further purified using the RNA-easy kit (QIAGEN). We confirmed equal expression of house-keeping genes in right and left halves using RT-PCR and then performed paired-end whole mRNA sequencing in LC Sciences (Houston, TX) followed by differential transcript analyses (>1 or <-1 log fold change; p < .05; q < .05; and FPKM >0.5 in 2/3 samples). The Mouse Genome Informatics and Online Mendelian Inheritance in Man databases as well as gnomAD constraint scores were used to prioritize differentially expressed transcripts. RESULTS There were 19 upregulated and 19 downregulated transcripts at E14.5 and 8 upregulated and 17 downregulated transcripts at E18.5 time-points. These differentially expressed transcripts were statistically significant and shown to be associated with craniofacial phenotypes in mouse models. These transcripts also have significant gnomAD constraint scores and are enriched in biological processes critical for embryogenesis. CONCLUSIONS We identified significant differential expression of transcripts between E14.5 and 18.5 murine right and left MxMn complexes. These findings when extrapolated to humans, they may provide a biological basis for facial asymmetry. Further experiments are required to validate these findings in murine models with craniofacial asymmetry.

[1]  M. Mikulewicz,et al.  Current Concepts and Challenges in the Treatment of Cleft Lip and Palate Patients—A Comprehensive Review , 2022, Journal of personalized medicine.

[2]  Bo Zhou,et al.  Physical, language, neurodevelopment and phenotype-genotype correlation of Chinese patients with Mowat-Wilson syndrome , 2022, Frontiers in Genetics.

[3]  Y. Zarate,et al.  Growth in individuals with SATB2‐associated syndrome , 2022, American journal of medical genetics. Part A.

[4]  B. Venkatesh,et al.  Discovery of a genetic module essential for assigning left–right asymmetry in humans and ancestral vertebrates , 2021, Nature Genetics.

[5]  D. MacArthur,et al.  Variant interpretation using population databases: Lessons from gnomAD , 2021, Human mutation.

[6]  L. Garavelli,et al.  Neurological Phenotype of Mowat-Wilson Syndrome , 2021, Genes.

[7]  D. Graf,et al.  Craniofacial Development: Neural Crest in Molecular Embryology , 2021, Head and Neck Pathology.

[8]  Liguo Wang,et al.  Novel risk factors for craniofacial microsomia and assessment of their utility in clinic diagnosis. , 2021, Human molecular genetics.

[9]  Ryan L. Collins,et al.  Author Correction: The mutational constraint spectrum quantified from variation in 141,456 humans , 2021, Nature.

[10]  A. V. Anagnostopoulos,et al.  Mouse Genome Database (MGD): Knowledgebase for mouse–human comparative biology , 2020, Nucleic Acids Res..

[11]  James A. Kadin,et al.  The mouse Gene Expression Database (GXD): 2021 update , 2020, Nucleic Acids Res..

[12]  Geo Pertea,et al.  GFF Utilities: GffRead and GffCompare , 2020, F1000Research.

[13]  Ryan L. Collins,et al.  The mutational constraint spectrum quantified from variation in 141,456 humans , 2020, Nature.

[14]  D. Grimes Making and breaking symmetry in development, growth and disease , 2019, Development.

[15]  Steven L Salzberg,et al.  Graph-based genome alignment and genotyping with HISAT2 and HISAT-genotype , 2019, Nature Biotechnology.

[16]  J. Vilo,et al.  g:Profiler: a web server for functional enrichment analysis and conversions of gene lists (2019 update) , 2019, Nucleic Acids Res..

[17]  Alan F. Scott,et al.  OMIM.org: leveraging knowledge across phenotype–gene relationships , 2018, Nucleic Acids Res..

[18]  J. Dai,et al.  Etiology and Pathogenesis of Hemifacial Microsomia , 2018, Journal of dental research.

[19]  Brent S. Pedersen,et al.  A map of constrained coding regions in the human genome , 2017, bioRxiv.

[20]  J. V. van Aalst,et al.  Genetic Advances in the Understanding of Microtia , 2016, Journal of Pediatric Genetics.

[21]  Xu Zhou,et al.  Genome-wide association study identifies multiple susceptibility loci for craniofacial microsomia , 2022 .

[22]  G. O’Keeffe,et al.  Zeb2: A multifunctional regulator of nervous system development , 2015, Progress in Neurobiology.

[23]  Alyssa C. Frazee,et al.  Ballgown bridges the gap between transcriptome assembly and expression analysis , 2015, Nature Biotechnology.

[24]  S. Salzberg,et al.  StringTie enables improved reconstruction of a transcriptome from RNA-seq reads , 2015, Nature Biotechnology.

[25]  François Schiettecatte,et al.  OMIM.org: Online Mendelian Inheritance in Man (OMIM®), an online catalog of human genes and genetic disorders , 2014, Nucleic Acids Res..

[26]  J. Tobias,et al.  Nodal Pathway Genes Are Down-regulated in Facial Asymmetry , 2014, The Journal of craniofacial surgery.

[27]  M. Gymrek,et al.  OTX2 Duplication Is Implicated in Hemifacial Microsomia , 2014, PloS one.

[28]  Lun-Jou Lo,et al.  Facial asymmetry: etiology, evaluation, and management. , 2011, Chang Gung medical journal.

[29]  Marcel Martin Cutadapt removes adapter sequences from high-throughput sequencing reads , 2011 .

[30]  Mark D. Robinson,et al.  edgeR: a Bioconductor package for differential expression analysis of digital gene expression data , 2009, Bioinform..

[31]  N. Byrd,et al.  Loss of Gbx2 results in neural crest cell patterning and pharyngeal arch artery defects in the mouse embryo. , 2005, Developmental biology.

[32]  B. D. de Vries,et al.  Disruption of the gene Euchromatin Histone Methyl Transferase1 (Eu-HMTase1) is associated with the 9q34 subtelomeric deletion syndrome , 2005, Journal of Medical Genetics.

[33]  M. Kimura,et al.  Validity of the Hfm transgenic mouse as a model for hemifacial microsomia. , 2002, The Cleft palate-craniofacial journal : official publication of the American Cleft Palate-Craniofacial Association.

[34]  S Peck,et al.  Skeletal asymmetry in esthetically pleasing faces. , 2009, The Angle orthodontist.