Abstract
Retinitis pigmentosa (RP) is a rare and heterogeneous group of inherited ocular diseases. However, the relationship between CACNA2D4 mutations and RP is not well understood. In this study, a Chinese autosomal recessive retinitis pigmentosa (arRP) pedigree was enrolled and targeted next-generation sequencing was employed for identifying the causative gene in the proband. These steps were followed by confirmatory Sanger sequencing and segregation analysis. RNA-sequencing (RNA-seq) data and semi-quantitative reverse transcription polymerase chain reaction analysis were then applied to examine the expressions in the human and mouse tissues. Novel compound heterozygous, deleterious missense variants of the CACNA2D4 gene, NM_172364.4: c.G955A (p.D319N) and c.A1822C (p.I608L), were identified in the arRP pedigree, co-segregating with the clinical phenotype in the patient. The CACNA2D4 protein is highly conserved among species. The CACNA2D4 mRNA expression showed the highest expression in the retina of humans and in the later four developmental stages/times of retinal tissues in mice, indicating its role in retina/eye functions and developments. This study is the first to identify novel compound heterozygous mutations c.G955A (p.D319N) and c.A1822C (p.I608L) in the CACNA2D4 gene. These might be disease-causing mutations, thereby extending the mutational spectra. The identification of pathogenic CACNA2D4 variants is expected to enhance our understanding of the genotype–phenotype correlations of arRP for disease diagnosis and genetic counseling. The relationship between the CACNA2D4 variants and diseases/phenotypes other than RP has also been reviewed and discussed in this paper.
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Abbreviations
- RP:
-
Retinitis pigmentosa
- arRP:
-
Autosomal recessive retinitis pigmentosa
- CACNA2D4:
-
Homo sapiens calcium voltage-gated channel auxiliary subunit alpha2delta 4 gene
- NGS:
-
Next-generation sequencing
- TGS:
-
Targeted next-generation sequencing
- RNA-seq:
-
RNA-sequencing
- RT-PCR:
-
Revere transcriptional-polymerase chain reaction
- ExAC:
-
The Exome Aggregation Consortium database
- HGMD:
-
The Human Gene Mutation Database
- RPKM:
-
Reads Per Kilobase of transcript per Million
- OMIM:
-
Online Mendelian Inheritance in Man
- NX:
-
Consensus normalized expression
- VGCC:
-
Voltage-gated calcium channels
- BWA:
-
Burrow–Wheeler Aligner
- ERG:
-
Electroretinogram
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Acknowledgements
This work was supported by the National Natural Science Foundation of China (31701087), the Special Training Program for Young Science and Technology Talents from Southwest Medical University (00031726), the Joint Research Foundation of Luzhou City and Southwest Medical University (2018LZXNYD-YL01).
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JF was in charge of the idea. J.F. and S.A. supervised the project. JC, JF, LZ, and CW performed DNA extraction, PCR, sequencing and data analysis. HL and QZ recruited the clinical patients and were in charge of the clinical assessment. JF and SK wrote the manuscript, and JF, SA and JC revised the manuscript. All authors read and approved the final manuscript.
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The study has the Ethical Committees approval granted by the Southwest Medical University. All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.
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Cheng, J., Zhou, Q., Fu, J. et al. Novel compound heterozygous missense variants (c.G955A and c.A1822C) of CACNA2D4 likely causing autosomal recessive retinitis pigmentosa in a Chinese patient. 3 Biotech 11, 208 (2021). https://doi.org/10.1007/s13205-021-02761-4
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DOI: https://doi.org/10.1007/s13205-021-02761-4