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Optimization of specific multiplex DNA primers to detect variable CLU genomic lesions in patients with Alzheimer’s disease

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Abstract

Recently, polymorphisms in the clusterin (CLU) or Apolipoprotein J (ApoJ) gene were reported to be involved in lipid metabolism, atherogenesis, and being associated with the risk of developing Alzheimer’s disease (AD). The influences from genetic variation has not been examined among Koreans. To screen genes with abnormal exon expression profiles, we developed PCR primer pairs for CLU gene. The new primer set can target specific regions of previously sequenced CLU gene. Primers were designed to target eight exon amplicons with flanking regions to optimize PCR amplification. One-hundred samples from clinically diagnosed Korean AD patients were selected for testing. We identified four single nucleotide polymorphisms (SNPs) in CLU through direct sequencing of the PCR products. These included three SNPs (rs7982, rs2279590 and rs3216167) that were previously reported variants in the SNP database (dbSNP), which could be found in NCBI. Interestingly, one new (NEW1) or extremely low-frequency mutation was found in more than one individual among the late-onset AD subjects. Our study suggests that CLU variants may also be an AD susceptibility factor among Koreans. Moreover, the findings of the study can be able to develop for simultaneous identify all of the genotypes of CLU mutation sites by DNA microarray PCR-based in the future.

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References

  1. Agnishwar, G., Jung, C., Mun, H.Y. & Park, H.G. PCRfree mutation detection of BRCA1 on a zip-code microarray using ligase chain reaction. Journal of Biochemical and Biophysical Method. 70, 897–902 (2008).

    Article  Google Scholar 

  2. Aronow, B.J., Lund, S.D., Brown, T.L., Harmony, J.A. & Witte, D.P. Apolipoprotein J expression at fluid-tissue interfaces: potential role in barrier cytoprotection. Proc. Natl. Acad. Sci. USA. 90, 725–729 (1993).

    Article  CAS  Google Scholar 

  3. Braskie, M.N. et al. Common Alzheimer’s disease risk variant within the CLU gene affects white matter microstructure in young adults. Journal of Neuroscienc. 31, 6764–6770 (2011).

    CAS  Google Scholar 

  4. Calero, M. et al. Apolipoprotein J (clusterin) and Alzheimer’s disease. Microsc. Res. Tech. 50, 305–315 (2000).

    Article  CAS  Google Scholar 

  5. Carrasquillo, M.M., Belbin, O., Hunter, T.A., Ma, L. & Bisceglio, G.D. Replication of CLU, CR1, and PICALM associations with Alzheimer disease. Archives of Neurolog. 67, 961–964 (2010).

    Google Scholar 

  6. Chen, L.H. et al. Polymorphisms of CR1, CLU and PICALM confer susceptibility of Alzheimer’s disease in a southern Chinese population. Neurobiol. Agin. 33, 211–217 (2012).

    Article  Google Scholar 

  7. Daimon, M. et al. Association of the clusterin gene polymorphisms with type 2 diabetes mellitus. Metabolis. 60, 815–822 (2011).

    Article  CAS  Google Scholar 

  8. Elias-Sonnenschein, L.S. et al. Genetic Loci Associated with Alzheimer’s Disease and Cerebrospinal Fluid Biomarkers in a Finnish Case-Control Cohort. PLoS One 8, e59676 (2013).

    Article  Google Scholar 

  9. Eva, B., Youn, Y.C., An, S.S.A. & Kim, S.Y. The genetics of Alzheimer’s disease. Clin. Interv. Agin. 9, 535–551 (2014).

    Google Scholar 

  10. Guerreiro, R.J. et al. Genetic variability in CLU and its association with Alzheimer’s disease. PLoS One 5, e9510 (2010).

    Article  Google Scholar 

  11. Harold, D. et al. Genome-wide association study identifies variants at CLU and PICALM associated with Alzheimer’s disease. Nat. Genet. 41, 1088–1093 (2009).

    Article  CAS  Google Scholar 

  12. Jun, G. et al. Meta-analysis confirms CR1, CLU, and PICALM as Alzheimer disease risk loci and reveals interactions with APOE genotypes. Arch Neurol. 67, 1473–1484 (2010).

    Article  Google Scholar 

  13. Kamboh, M.I. et al. Association of CLU and PICALM variants with Alzheimer’s disease. Neurobiol. Agin. 33, 518–521 (2012).

    Article  CAS  Google Scholar 

  14. Komatsu, M. et al. Genetic association between clusterin polymorphisms and Alzheimer’s disease in a Japanese population. Psychogeriatric. 11, 14–18 (2011).

    Article  Google Scholar 

  15. Lambert, J.C. et al. Genomewide association study identifies variants at CLU and CR1 associated with Alzheimer’s disease. Nat. Genet. 41, 1094–1099 (2009).

    Article  CAS  Google Scholar 

  16. Lancaster, T.M. et al. Neural hyperactivation in carriers of the Alzheimer’s risk variant on the clusterin gene. European Neuropsychopharmacolog. 21, 880–884 (2011).

    Article  CAS  Google Scholar 

  17. May, P.C. et al. Dynamics of gene expression for a hippocampal glycoprotein elevated in Alzheimer’s disease and in response to experimental lesions in rat. Neuro. 5, 831–839 (1990).

    Article  CAS  Google Scholar 

  18. Miyashita, A. et al. SORL1 is genetically associated with late-onset Alzheimer’s disease in Japanese, Koreans and Caucasians. PLoS One 8, e58618 (2013).

    Article  Google Scholar 

  19. Song, J.Y., Park, H.G., Jung, S.O. & Park, J. Diagnosis of HNF-1alpha mutations on a PNA zip-code microarray by single base extension. Nucleic Acids Res. 33, p. e19 (2005).

    Article  Google Scholar 

  20. Swaroop, A., Chew, E.Y., Rickman, C.B. & Abecasis, G.R. Unraveling a multifactorial late-onset disease: from genetic susceptibility to disease mechanisms for age-related macular degeneration. Annu. Rev. Genomics Hum. Genet. 10, 19–43 (2009).

    Article  CAS  Google Scholar 

  21. Sweet, R.A. et al. Effect of Alzheimer’s disease risk genes on trajectories of cognitive function in the Cardiovascular Health Study. Am. J. Psychiatr. 169, 954–962 (2012).

    Article  Google Scholar 

  22. Thambisetty, M. et al. Plasma clusterin concentration is associated with longitudinal brain atrophy in mild cognitive impairment. NeuroImag. 59, 212–217 (2012).

    Article  CAS  Google Scholar 

  23. Tyagi, S., Bratu, D.P. & Kramer, F.R. Multicolor molecular beacons for allele discrimination. Nat. Biotechnol. 16, 49–53 (1998).

    Article  CAS  Google Scholar 

  24. Untergasser, A. et al. Primer3Plus, an enhanced web interface to Primer3. Nucleic Acids Res. 35, W71–W74 (2007).

    Article  Google Scholar 

  25. Wang, D.G. et al. Large-scale identification, mapping, and genotyping of single-nucleotide polymorphism in the human genome. Scienc. 280, 1077–1082 (1998).

    Article  CAS  Google Scholar 

  26. Wu, Z.C., Yu, J.T., Li, Y. & Tan, L. Clusterin in Alzheimer’s disease. Advances in Clinical Chemistr. 56, 155–173 (2012).

    Article  CAS  Google Scholar 

  27. Yim, S.C., Park, H.G., Chang, H.N. & Cho, D.Y. Array-based mutation detection of BRCA1 using direct probe/target hybridization. Anal. Biochem. 337, 332–337 (2005).

    Article  CAS  Google Scholar 

  28. Yu, J.T. et al. Implication of CLU gene polymorphisms in Chinese patients with Alzheimer’s disease. Clin. Chim. Act. 411, 1516–1519 (2010).

    Article  CAS  Google Scholar 

  29. Zabarovsky, E.R. et al. Restriction site tagged (RST) microarrays: a novel technique to study the species composition of complex microbial systems. Nucleic Acids Res. 31, e95 (2003).

    Article  Google Scholar 

  30. Zhang, S. et al. CLU rs2279590 polymorphism contributes to Alzheimer’s disease susceptibility in Caucasian and Asian populations. J. Neural. Transm. 122, 433–439 (2015).

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Bionano Technology & Gachon Medical Research Institute, Gachon University, 1342 Sungnam-daero, Sujung-gu, Seongnam-si, Gyeonggi-do, 461-701, Korea

    Vo Van Giau & Seong Soo A. An

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  1. Vo Van Giau
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  2. Seong Soo A. An
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Correspondence to Seong Soo A. An.

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Van Giau, V., An, S.S.A. Optimization of specific multiplex DNA primers to detect variable CLU genomic lesions in patients with Alzheimer’s disease. BioChip J 9, 278–284 (2015). https://doi.org/10.1007/s13206-015-9306-8

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  • DOI: https://doi.org/10.1007/s13206-015-9306-8

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