Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter to the Editor
  • Published:

Mitochondrial DNA minisatellites as new markers for the quantitative determination of hematopoietic chimerism after allogeneic stem cell transplantation

Leukemia volume 21, pages 369–373 (2007)Cite this article

The monitoring of chimerism after allogeneic stem cell transplantation (SCT) is indispensable because it can enable a prompt therapeutic intervention for early relapse or rejection. Moreover, chimerism analysis may be a valuable alternative in monitoring minimal residual disease.1 The short tandem repeat (STR) markers have been increasingly used because they are highly polymorphic between individuals, and the STR assay produces quantitative results using fluorescence-labeled primers and a capillary electrophoresis system. However, a recent study highlighted potential problems of the STR assay showing poor precision and a relatively lower sensitivity, particularly when the target DNA content is low and degraded.2 In addition, there were no common guidelines for interpreting the STR assays with respect to the clinical response. A study on the imprecision of seven STRs (D7S820, D8S1179, D16S539, D18S51, D21S11, TH01 and TPOX) using mixed blood cells at various concentrations showed that the imprecision ranged from 5.4% for TPOX to 12% for D7S820, and it was inversely related to the proportion of the concentration of the cell mixtures. A careful approach and consideration will be needed because imprecision can potentially be high, particularly with a lower proportion of donor cells.2 To circumvent these problems, six mitochondrial DNA (mtDNA) minisatellite (mtMS) markers based on our recent report3 and other publication regarding mtDNA genetic diversity among Korean4 were developed to monitor the extent of donor cell engraftment in SCT recipients and to evaluate their informativeness and sensitivity compared with the nuclear DNA markers.

Nucleated animal cells contain approximately 1000 mitochondria with each containing 2–10 mtDNA. Therefore, a typical differentiated cell has approximately 3000 copies of mtDNA.5 The mtDNA control region includes two hypervariable regions (HV1 and HV2), so called because of their high incidence of nucleotide variations. Many mtDNA length heteroplasmies (mtMS) are localized in the HV2 homopolymeric C (poly-C) tract, which lies between nucleotide positions (np) 303–315. In this paper, the length heteroplasmic sequences are abbreviated as 303CCCCCCCTCCCCC315 as 7CT5C (303 poly-C). Another poly-C tract variant is located between np 16184–16193 (16184CCCCCTCCCC16193, 5CT4C) (16184 poly-C) in the HV1 region. During the construction of the Korean mtDNA database, all Koreans were found to display length heteroplasmy in both HV1 and HV2, and a high level of length heteroplasmy in the poly-C-tract of HV2. Poly-C length heteroplasmy profiling in hematopoietic tissues after allogeneic SCT may also be used to quantitatively analyze chimerism. Fifty-five to 60% of genetic alterations in CD34+ cells, T cells and B cells were length changes in the homopolymeric C tract at np 303–315 in HV2.6 Therefore, heterogeneity of the mtDNA sequence in normal adult CD34+ cells might have implications such as, a natural genetic ‘marker’ of hematopoietic progenitors and stem cells to estimate the number of active stem cells in the steady state.7 The patterns of mutations that have accumulated over time along the transmission of the mtDNA lineage give rise to differences between individuals, and have become the basis for forensic discrimination.4

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1

References

  1. Fernandez-Aviles F, Urbano-Ispizua A, Aymerich M, Colomer D, Rovira M, Martinez C et al. Serial quantification of lymphoid and myeloid mixed chimerism using multiplex PCR amplification of short tandem repeat-markers predicts graft rejection and relapse, respectively, after allogeneic transplantation of CD34+ selected cells from peripheral blood. Leukemia 2003; 17: 613–620.

    Article  CAS  Google Scholar 

  2. Kong SY, Ki CS, Kim HJ, Lee KO, Bae JC, Kim SH et al. Evaluation of imprecision for analysis of short tandem repeats by use of mixed blood cells in variable concentrations. Clin Chem 2004; 50: 2193–2195.

    Article  CAS  Google Scholar 

  3. Shin MG, Levin BC, Kim HJ, Kim HR, Lee IK, Cho D et al. Profiling of length heteroplasmies in the human mitochondrial DNA control regions from blood cells in the Korean population. Electrophoresis 2006; 27: 1331–1340.

    Article  CAS  Google Scholar 

  4. Lee HY, Yoo JE, Park MJ, Chung U, Shin KJ . Mitochondrial DNA control region sequences in Koreans: identification of useful variable sites and phylogenetic analysis for mtDNA data quality control. Int J Legal Med 2006; 120: 5–14.

    Article  Google Scholar 

  5. Naviaux RK . Mitochondrial DNA disorders. Eur J Pediatr 2000; 159 (Suppl 3): S219–S226.

    Article  CAS  Google Scholar 

  6. Ogasawara Y, Nakayama K, Tarnowka M, McCoy Jr JP, Kajigaya S, Levin BC et al. Mitochondrial DNA spectra of single human CD34+ cells, T cells, B cells, and granulocytes. Blood 2005; 106: 3271–3284.

    Article  CAS  Google Scholar 

  7. Shin MG, Kajigaya S, Tarnowka M, McCoy Jr JP, Levin BC, Young NS . Mitochondrial DNA sequence heterogeneity in circulating normal human CD34 cells and granulocytes. Blood 2004; 103: 4466–4477.

    Article  CAS  Google Scholar 

  8. Butler JM, Levin BC . Forensic applications of mitochondrial DNA. Trends Biotechnol 1998; 16: 158–162.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This study was supported by Grant no. R01-2004-000-10540-0 from the Basic Research Program of the Korea Science & Engineering Foundation (to Myung-Geun Shin), by a grant from the Korea Health 21 R&D project, Ministry of Health & Welfare, Republic of Korea (01-PJ10-PG6-01GN16-0005) (to Hyeoung-Joon Kim).

Author information

Authors and Affiliations

  1. Department of Laboratory Medicine, Chonnam National University Hwasun Hospital, Hwasun, Korea

    M-G Shin

  2. Genome Research Center for Hematopoietic Disease, Chonnam National University Medical School and Chonnam National University Hwasun Hospital, Hwasun, Korea

    H-J Kim & H-R Kim

  3. Department of Pediatrics, Chonnam National University Medical School and Chonnam National University Hwasun Hospital, Hwasun, Korea

    Il-K Lee, H Kook, D Cho, S-J Kee, J-H Shin, S-P Suh & D-W Ryang

Authors
  1. M-G Shin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. H-J Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. H-R Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Il-K Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. H Kook
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. D Cho
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. S-J Kee
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. J-H Shin
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. S-P Suh
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. D-W Ryang
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Supplementary Information accompanies the paper on the Leukemia website (http://www.nature.com/leu)

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Shin, MG., Kim, HJ., Kim, HR. et al. Mitochondrial DNA minisatellites as new markers for the quantitative determination of hematopoietic chimerism after allogeneic stem cell transplantation. Leukemia 21, 369–373 (2007). https://doi.org/10.1038/sj.leu.2404502

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.leu.2404502

Search

Advanced search

Quick links