Cell Stem Cell. 2015 Jan 8;16(1):33-8. doi: 10.1016/j.stem.2014.11.003.

SCNT-derived ESCs with mismatched mitochondria trigger an immune response in allogeneic hosts.

Deuse T1, Wang D2, Stubbendorff M2, Itagaki R2, Grabosch A2, Greaves LC3, Alawi M4, Grünewald A5, Hu X2, Hua X2, Velden J6, Reichenspurner H7, Robbins RC8, Jaenisch R9, Weissman IL10, Schrepfer S11.
  • 1TSI Laboratory, University Heart Center Hamburg, Martinistrasse 52, 20246 Hamburg, Germany; Cardiovascular Research Center Hamburg (CVRC) and DZHK (German Center for Cardiovascular Research), partner site Hamburg/Kiel/Luebeck, Martinistrasse 52, 20246 Hamburg, Germany; Cardiovascular Surgery, University Heart Center Hamburg, Martinistrasse 52, 20246 Hamburg, Germany.
  • 2TSI Laboratory, University Heart Center Hamburg, Martinistrasse 52, 20246 Hamburg, Germany; Cardiovascular Research Center Hamburg (CVRC) and DZHK (German Center for Cardiovascular Research), partner site Hamburg/Kiel/Luebeck, Martinistrasse 52, 20246 Hamburg, Germany.
  • 3Newcastle University Centre for Brain Ageing and Vitality, Newcastle University, Newcastle upon Tyne, NE2 4HH, UK.
  • 4Bioinformatics Service Facility, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, 20246 Hamburg, Germany; Heinrich-Pette Institute, Leibniz Institute for Experimental Virology, Virus Genomics, Martinistrasse 52, 20246 Hamburg, Germany.
  • 5Wellcome Trust Centre for Mitochondrial Research, Institute of Neuroscience, Newcastle University, Newcastle upon Tyne, NE2 4HH, UK.
  • 6Department of Nephropathology, Institute of Pathology, University Hospital Erlangen, Maximiliansplatz 2, 91054 Erlangen, Germany.
  • 7Cardiovascular Research Center Hamburg (CVRC) and DZHK (German Center for Cardiovascular Research), partner site Hamburg/Kiel/Luebeck, Martinistrasse 52, 20246 Hamburg, Germany; Cardiovascular Surgery, University Heart Center Hamburg, Martinistrasse 52, 20246 Hamburg, Germany.
  • 8Stanford Cardiovascular Institute and Department of Cardiothoracic Surgery, Stanford University School of Medicine, 300 Pasteur Drive, Stanford, CA 94305, USA.
  • 9Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology, 9 Cambridge Center, Cambridge, MA 02142, USA.
  • 10Department of Developmental Biology, Stanford Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, 300 Pasteur Drive, Stanford, CA 94305, USA.
  • 11TSI Laboratory, University Heart Center Hamburg, Martinistrasse 52, 20246 Hamburg, Germany; Cardiovascular Research Center Hamburg (CVRC) and DZHK (German Center for Cardiovascular Research), partner site Hamburg/Kiel/Luebeck, Martinistrasse 52, 20246 Hamburg, Germany; Stanford Cardiovascular Institute and Department of Cardiothoracic Surgery, Stanford University School of Medicine, 300 Pasteur Drive, Stanford, CA 94305, USA. Electronic address: schrepfer@stanford.edu.

 

Abstract

The generation of pluripotent stem cells by somatic cell nuclear transfer (SCNT) has recently been achieved in human cells and sparked new interest in this technology. The authors reporting this methodical breakthrough speculated that SCNT would allow the creation of patient-matched embryonic stem cells, even in patients with hereditary mitochondrial diseases. However, herein we show that mismatched mitochondria in nuclear-transfer-derived embryonic stem cells (NT-ESCs) possess alloantigenicity and are subject to immune rejection. In a murine transplantation setup, we demonstrate that allogeneic mitochondria in NT-ESCs, which are nucleus-identical to the recipient, may trigger an adaptive alloimmune response that impairs the survival of NT-ESC grafts. The immune response is adaptive, directed against mitochondrial content, and amenable for tolerance induction. Mitochondrial alloantigenicity should therefore be considered when developing therapeutic SCNT-based strategies.

PMID: 25465116

 

 

Supplement:

Mitochondrial Immunogenicity in Embryonic Stem Cells

Congestive heart failure is the leading cause of death in the United States and Western Europe. Therapeutic options for end-stage heart failure currently include heart transplantation and assist device implantation, but transplantation of stem cell-derived cell therapy to regenerate the damaged myocardium possesses great potential for future strategies. Skeletal myoblasts and a variety of different types of adult stem cells have been investigated clinically for the regeneration of injured myocardium. However, there is little reproducible evidence that any of these cells differentiate into cardiomyocytes or significantly contribute to the regrowth of heart tissue.

Previously, we have investigated transplantation of adult mesenchymal stem cells (MSCs) for the treatment of heart failure following myocardial infarction in mice and have observed an improvement in myocardial function1. Because the survival of cellular MSC grafts was limited, the improvement in myocardial function could be unambiguously attributed to the cell’s paracrine effects1.

Embryonic stem cells (ESC) theoretically hold the most promise for the treatment of heart failure since they are capable of differentiating into cardiomyocytes owing to their naïve pluripotency. The Transplant and Stem Cell Immunobiology Lab (TSI) at the University Heart Center Hamburg (Germany) is aiming at investigating how immune rejection and non-immunological factors such as ischemia and inflammation affect survival and differentiation of ESC grafts. In this context, the generation of autologous pluripotent stem cells for specific patient-matched therapies seems to be particularly appealing. The first embryonic stem cells created by somatic cell nucleus transfer (SCNT) of human origin were generated in 2013 from a fetal fibroblast2. SCNT involves transferring the nucleus of an adult somatic cell into an egg cell from which the nucleus had previously been removed. Embryonic stem cells generated with this technology might be more promising than iPS cells, which are somatic cells induced to regain stemness but exhibiting additional iPS-specific immune features3. Although SCNT-ESC transplantation could at first sight circumvent immune rejection provided that one patient both supplied the nucleus and also received the stem cell graft, the authors worried that SCNT-derived cells might still provoke an immune reaction1. Even though SCNT-ESCs possess the identical nuclear genome of the nucleus donor, they still contain mitochondrial DNA (mtDNA) originating from the egg donor1, 2. Indeed, the experiments revealed that proteins derived from mismatched mtDNA induced an immune response when SCNT-ESCs were transplanted back into the nucleus donor mouse strain. The publication demonstrates that only two single nucleotide polymorphisms (SNPs) in the mtDNA of the SCNT-ESCs were sufficient to cause an adaptive immune response, which, however, was amenable for tolerance induction1. The SNPs were located within the mt-Co3 and mt-Cytb genes and caused non-synonymous amino acid substitutions in the related proteins. These two SNPs were the only differences in the whole cellular DNA between the SCNT-ESCs and the recipients and both variation are usual in the mouse1, 4.

image description

Figure 1. The artwork depicts the ratio of nuclear (blue) and mitochondrial (red) DNA in a confocal 3D image of an oocyte (taken by Wang). Deuse, who is the grand-grand-grand son of the German poet Christian Wagner, contributed the poem summarizing the main findings of the manuscript.

 

Figure 1 summarizes and simplifies the main findings in a poem. Until this study appeared, the mitochondrial mismatch idea had never been tested for SCNT-ESCs.

Although rejection of SCNT-ESCs by the nucleus donor was only observed in a mouse model, the heterogeneity in human mitochondria with 2066 SNPs is substantially higher than in mouse5. A recent report indicates that each individual holds an average of 25.3 variants of SNPs compared to the human reference sequence6. For the therapeutic use of SCNT-derived cells, this finding is vitally important. The immune reactions reported in this paper will need to be considered if clinicians aim to use SCNT-derived cells for human therapy. Sequencing and matching of mtDNA could be performed to increase the margin of safety attributed to this technology. Also, egg donors directly related to the patient such as mother or sister could provide mtDNA matched oocytes for the SCNT.

 

References

  1. Deuse T, Wang D, Stubbendorff M, Itagaki R, Grabosch A, Greaves LC, Alawi M, Grunewald A, Hu X, Hua X, Velden J, Reichenspurner H, Robbins RC, Jaenisch R, Weissman IL, Schrepfer S. Scnt-derived escs with mismatched mitochondria trigger an immune response in allogeneic hosts. Cell stem cell. 2015;16:33-38
  2. Tachibana M, Amato P, Sparman M, Gutierrez NM, Tippner-Hedges R, Ma H, Kang E, Fulati A, Lee HS, Sritanaudomchai H, Masterson K, Larson J, Eaton D, Sadler-Fredd K, Battaglia D, Lee D, Wu D, Jensen J, Patton P, Gokhale S, Stouffer RL, Wolf D, Mitalipov S. Human embryonic stem cells derived by somatic cell nuclear transfer. Cell. 2013;153:1228-1238
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  4. Sherry ST, Ward MH, Kholodov M, Baker J, Phan L, Smigielski EM, Sirotkin K. Dbsnp: The ncbi database of genetic variation. Nucleic Acids Res. 2001;29:308-311
  5. Goios A, Pereira L, Bogue M, Macaulay V, Amorim A. Mtdna phylogeny and evolution of laboratory mouse strains. Genome research. 2007;17:293-298
  6. Ridge PG, Maxwell TJ, Foutz SJ, Bailey MH, Corcoran CD, Tschanz JT, Norton MC, Munger RG, O’Brien E, Kerber RA, Cawthon RM, Kauwe JS. Mitochondrial genomic variation associated with higher mitochondrial copy number: The cache county study on memory health and aging. BMC bioinformatics. 2014;15 Suppl 7:S6

 

 

 

 

 

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