OVERVIEW
Transplantation Between Monozygotic Twins: How Identical Are They? Elizabeth Day,1,5 Patrick K. Kearns,1 Craig J. Taylor,2,3 and J. Andrew Bradley 3,4 Abstract. Advances in developmental biology have shown that monozygous twins may not be as phenotypically identical as once believed, and the mechanisms responsible for such differences are now becoming clearer. Whether such phenotypic differences are capable of triggering graft rejection of an organ transplanted between identical twins remains unknown but the risks seem low, and long-term transplant outcome is excellent. Available evidence to guide immunosuppressive therapy in this setting is limited but a prudent approach would include the use of steroids together with a calcineurin inhibitor after transplantation. However, once the inevitable inflammatory response associated with transplant surgery has resolved, cautious reduction and eventually withdrawal of immunosuppression should be possible. Keywords: Twin, Transplantation, Genetic. (Transplantation 2014;98: 485Y489)
he first successful renal transplant was carried out between ‘‘identical’’ twins almost six decades ago and since that landmark case over 200 kidney transplants between identical twins have been recorded in the United States (1, 2). The early cases of renal transplantation between identical twins were carried out without immunosuppression by necessity because at the time safe and effective immunosuppression was not available. In subsequent years, successful transplantation of kidneys between identical twins has continued with the use of minimal or no immunosuppression (3Y5).
T
The authors declare no funding or conflicts of interest. 1 Department of Surgery, Southern General Hospital, Glasgow, United Kingdom. 2 Histocompatibility and Immunogenetics Laboratory, Cambridge University Teaching Hospitals NHS Foundation Trust, Addenbrooke’s Hospital, Cambridge, United Kingdom. 3 NIHR Biomedical Research Centre, Cambridge, United Kingdom. 4 Department of Surgery, University of Cambridge Clinical School, Addenbrooke’s Hospital, Cambridge, United Kingdom. 5 Address correspondence to: Elizabeth Day, Ph.D., Department of Surgery, Southern General Hospital, Glasgow, Lanarkshire, G51 4TF, United Kingdom. E-mail: [email protected] E.D. conceived and drafted, and helped revise the review. P.K.K. contributed to first and subsequent drafts of the review. C.J.T. participated in editing the first draft and subsequent drafts of the review. J.A.B. contributed to the first and subsequent drafts of the review. All the authors have approved the article for publication. Supplemental digital content (SDC) is available for this article. Direct URL citations appear in the printed text, and links to the digital files are provided in the HTML text of this article on the journal’s Web site (www.transplantjournal.com). Received 17 December 2013. Revision requested 24 February 2014. Accepted 1 May 2014. Copyright * 2014 by Lippincott Williams & Wilkins ISSN: 0041-1337/14/9805-485 DOI: 10.1097/TP.0000000000000274
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Interestingly, however, our current understanding of the molecular basis of development and the in utero environment suggests that monozygous (MZ) twins may not be as identical as previously believed. The clinical relevance of these differences in organ transplantation is unclear, and there is little study data pertaining to necessity of immunosuppressive therapy in identical twin transplantation (6). We provide a brief review of the current status of MZ twin transplantation and examine the practical and theoretical implications of current genetic, epigenetic, and immunologic research, with a particular focus on renal transplantation.
HOW IDENTICAL ARE IDENTICAL TWINS? MZ twins arise from a single ovum fertilized by a single sperm. Traditionally they are considered genetically identical but phenotypically MZ twins do show differences. Historically, these phenotypic differences have been attributed to environmental influences, including effects on epigenetic reprogramming (7). The interaction between genes and modulating environmental stimuli begins early in utero and lifestyle differences give rise to genetic variation that increases with age (epigenetic drift). Genome-wide and single locus DNA methylation patterns leads to epigenetic variability in gene promoters that cause differences in transcriptional regulation and complex phenotypes (8). Furthermore, in the last decade, a number of studies have shown that MZ twins can and do differ genetically, with changes affecting single base pairs, and rarely at larger genomic regions and whole chromosomes (9Y12). This phenomenon is related to the concept of somatic variation, whereby the genetic and epigenetic contents of cells in a single individual differ. Somatic variation and MZ twin discordance arises through three main mechanisms: somatic mosaicism, chimerism, and www.transplantjournal.com
Copyright © 2014 Lippincott Williams & Wilkins. Unauthorized reproduction of this article is prohibited.
485
486
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epigenetic alterations accumulated throughout life (epigenetic drift) (Fig. 1). Somatic mosaicism is not a new concept and is vital for the normal functioning of the immune system with the controlled reshuffling of immunoglobulin and T-cell receptor genes; MZ twins are confronted with different foreign antigens and hence their T-cell reservoirs will vary. However, the presence and prevalence of somatic mosaicism in other tissues is a more recent observation. Somatic mosaics originate
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from a single zygote (the earliest stage of an embryo development) but contain cells with differing genetic content. Somatic mosaics can arise through a number of mechanisms, including mutations and structural changes occurring after the zygote stage, heteroplasmy (unequal division of mitochondrial DNA within the cellular cytoplasm) and uniparental disomy (where both copies of a chromosome or genomic region are inherited from a single parent) (13). In addition, development and maintenance of any single cell within an
FIGURE 1. Mechanisms of somatic variation. A, somatic mosaicism. Genetic changes occur spontaneously in the early stages of embryo development. These changes are propagated during the development of the fetus and result in a genetic mosaicVtissues contain cells with varying genetic content (gray shades). B, chimerism. Novel genetic material is introduced from an exogenous source. This source may be maternal or from a synchronous fetus. Cells or DNA fragments are believed to pass into the fetal circulation across the placenta or through the amniotic fluid. These cells or DNA must then integrate and propagate to contribute to the chimera. C, epigenetic drift. Epigenetic changes occur over time and, as with somatic mosaicism and chimerism, are propagated to result in a mix of expression patterns within a tissue.
Copyright © 2014 Lippincott Williams & Wilkins. Unauthorized reproduction of this article is prohibited.
Day et al.
* 2014 Lippincott Williams & Wilkins
TABLE 1.
487
Case reports of solid organ and composite tissue transplants between identical twins
Kidney A single case after graft failure from an earlier transplant. Three cases. Primary renal disease recurred in all cases after discontinuation of immunosuppression. Case series of simultaneous pancreas-kidney transplant including one identical twin pair. A single case of a simultaneous pancreas-kidney transplant in an insulin dependent diabetic transplanted after the recognition of renal failure. Liver Two cases requiring transplantation for hepatitis C related cirrhosis. Two cases requiring transplantation for hepatocellular carcinoma and a carcinoid tumor. A single case requiring transplantation for hepatitis C related cirrhosis. Small Bowel A single case of transplantation after gastrointestinal necrotizing vasculitis. A single case of transplantation of terminal ileum after resection for a mid-gut volvulus. Skin graft testing was performed before transplantation. A single case with a 4-yr follow up. Other Ovarian. Case seriesVseven ovarian transplants, five pregnancies at the time of follow up. Composite Tissue: A single case of a deep inferior epigastric flap donated for breast reconstruction.
Immunosuppression
Reference
Oral steroids and mofetil mycophenolate. Discontinued at 12, 15, and 91 mo after transplantation. Maintained on low-dose cyclosporine.
(6) (4)
Azathioprine for 1 mo and maintenance on low dose cyclosporine.
(34)
V
(35)
V
(36)
(33)
Steroid therapy for 1 mo
(37)
Cyclophosphamide initially and maintenance on oral steroids. V
(38) (39)
V
(40)
V
(41)
V
(42)
Selected case reports published over the last 15 years (1998Y2014) of solid organ and composite tissue transplantation between identical twins. The second column indicates whether immunosuppression was used, a (V) indicates that no immunosuppression was used.
organism is stochastic; genetic and epigenetic modifications will continue throughout life in response to the environment and ageing. Indeed, studies of MZ twins have shown that both epigenetic and genetic changes occur in response to the in utero environment and increase with age (14Y17). A chimera originates from the mixing of cells from individuals originating from two distinct zygotes. Cells must mix, implant and survive in the new organism. This may occur ‘‘naturally’’ in utero, maternal cells entering the fetal circulation, and conversely during pregnancy, whereby fetal cells enter the maternal circulation (18, 19). It may also take place between embryos in multiple pregnancies (20). The extent of this phenomenon is unknown but has been seen in hematopoietic tissue as well as hepatocytes, myocytes and islet A cells (20Y22). Indeed, in one recent review, the author suggests ‘‘that a shift is needed from the conventional paradigm of ‘self versus other’ to a view of the normal ‘self ’ as constitutively chimeric’’ (20). Iatrogenic chimerism occurs transiently after blood transfusion or more permanently after tissue and organ transplantation (23, 24). The full extent of somatic mosaicism or chimerism in the MZ twin population has not been studied: case report data tends to focus on overtly discordant twins. Indeed, it is technically difficult to estimate the prevalence of such
variation in the larger non-twin population, and recent studies have only begun to assess the level of genomic and epigenomic diversity within a single individual (17, 25). However, with the construction of large databases containing phenotypic and genetic information from twins, our understanding of normal twin variation is set to expand (15). The twin model is particularly powerful method for assessing somatic variation, and as such, the understanding we gain from studying twin-twin variation may well prove useful in the future to the transplant team (26). Zygosity Testing A comprehensive review of twin zygosity testing is beyond the scope of this article, but it remains an important consideration in the context of organ transplantation between twins. In general monochorionicty is a good screen for monozygosity (27). Standard human leukocyte antigen (HLA) typing is not sufficient alone to determine monozygosity because 25% of dizygotic twins will match at all HLA loci (28). More extensive haplotyping, such as that used in bone marrow transplantation, may add strength to choronicity, as would analysis of additional genetic markers, such as short tandem repeats, used in DNA fingerprinting. However, genetic markers, as with any other genomic region, may
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be affected by somatic variation, and therefore any anomalies detected by genetic testing must be considered with this in mind. The complexity of zygosity testing arises from many of the concepts that have been discussed in the preceding paragraphs; a review by Machin (27) offers a thorough discussion of the subject. In the context of organ and tissue transplantation, there is perhaps scope for more ‘‘functional’’ assays, akin to skin grafting seen in the early transplantations (1). Of course, choosing an appropriate substrate for these investigations would require substantial research, and the use of renal tissue would pose a practical challenge.
IMPLICATIONS OF MZ TWIN DISCORDANCE IN TRANSPLANTATION Clinically, MZ twin discordance has been described for single gene disorders, such as hemophilia B and Duchenne Muscular Dystrophy, as well as in complex conditions, such as multiple sclerosis and congenital heart disease (10, 29, 30). There is also a high prevalence of discordance for autoimmune conditions, such as systemic lupus erythematosus and rheumatoid arthritis: somatic and epigenetic variation between MZ twins identified by next generation (DNA) sequencing technologies may hint at potential immunologic discordance between MZ twins and hence have implications for transplantation (31, 32). Although the mechanisms through which this discordance arises are becoming apparent, translating this information into clinical decisions is a challenge. In the context of MZ twin transplantation, it is the requirement and degree of immunosuppression necessary that remains an uncertainty.
IMMUNOSUPPRESSION IN MZ TRANSPLANTATION Immunosuppressive regimens must balance the risks of immunosuppression, namely, infection, malignancy, and other specific side effects of agents, with the benefits of preventing graft rejection and disease recurrence. To ascertain the evidence base for immunosuppression in transplants performed between identical twins, we searched the literature to identify studies and reports pertaining to MZ twin organ transplantation (Appendix SI, SDC, http://links.lww.com/TP/B17). The most comprehensive analysis of MZ twin transplant cases was compiled by Krishnan and colleagues who identified close to 200 MZ twin renal transplants in the United States between 1987 and 2006. They reported that 71% of cases were receiving immunosuppressive therapy at the time of discharge from hospital (61% received steroids and 31% calcineurin inhibitors), falling to 34% 1 year and 33% 3 years after transplantation. At 1 year, 21% of recipients were still on calcineurin inhibitors, and 27% were still on steroids. Interestingly, graft function was superior in recipients who were not maintained on immunosuppression. There are limited data available pertaining to rejection rates in MZ twin transplantation to provide evidence-based guidance on the use of immunosuppression. Kessaris and colleagues (5), who identified 132 living-donor identical twin transplants that took place between 1988 and 2004 in the United Kingdom and United States, recorded similar observations to those of Krishnan et al: a large proportion of the
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patients remained on immunosuppression and there was no significant difference in graft survival between those on immunosuppression and those not. The findings from these reports suggest that renal transplantation between MZ twins can be performed safely without long-term immunosuppression. Successful living-donor liver and small bowel transplants have also been carried out without immunosuppression in MZ twins (Table 1) (4, 6, 33Y42).
DISCUSSION AND CONCLUSION It has long been observed that graft rejection is not solely a function of immune homology (43). Trauma, ischemia and reperfusion during organ transplantation stimulates cytokine release, activating immune responses and contributing to graft rejection (44). The presence of autoantibodies to cryptic tissue antigens (e.g., vimentin) associated with kidney and heart allograft rejection indicate that tissue damage may be an important initiator of graft rejection regardless of genetic identity (45). For this reason, perioperative steroids are an important component of immunosuppressive regimens and one that should most likely remain in MZ twin transplantation (2). Twin and HLA-identical sibling transplantation also carries additional considerations, not related to immunosuppression, such as distinct emotional consequences and disease recurrence: one of the most common reasons for graft loss in HLA-identical sibling transplants (46Y48). When planning renal transplantation between apparent MZ twins, zygosity testing using multilocus probes should be carried out to confirm twins are indeed monozygotic. Advances in developmental biology have clearly shown that MZ twins may not be as identical as once believed, and the mechanisms responsible for such differences are now becoming clearer. To what extent such differences are capable of triggering graft rejection remains unknown, but the risks seem low, and long-term transplant outcome is excellent. Available evidence to guide immunosuppressive is limited but a prudent approach would include the use of steroids together with a calcineurin inhibitor after transplantation. However, once the inflammatory response associated with transplant surgery has resolved, cautious reduction and eventual withdrawal of immunosuppression should be possible, thereby avoiding the long-term side effects of immunosuppression, including the detrimental effects of calcineurin inhibitors on graft function. REFERENCES 1.
2. 3.
4. 5.
Merrill JP, Murray JE, Harrison JH, et al. Successful homotransplantation of the human kidney between identical twins. J Am Med Assoc 1956; 160: 277. Krishnan N, Buchanan PM, Dzebisashvili N, et al. Monozygotic transplantation: concerns and opportunities. Am J Transplant 2008; 8: 2343. Gumprich M, Woeste G, Kohlhaw K, et al. Living related kidney transplantation between identical twins. Transplant Proc 2002; 34: 2205. Kim YK, Yoon HE, Kim SH, et al. Long-term follow-up of three identical twin transplant recipients without maintenance immunosuppressive therapy. Nephrology (Carlton) 2008; 13: 447. Kessaris N, Mukherjee D, Chandak P, et al. Renal transplantation in identical twins in United States and United Kingdom. Transplantation 2008; 86: 1572.
Copyright © 2014 Lippincott Williams & Wilkins. Unauthorized reproduction of this article is prohibited.
Day et al.
* 2014 Lippincott Williams & Wilkins
6.
7.
8. 9. 10.
11.
12.
13.
14.
15.
16.
17.
18. 19.
20. 21.
22.
23.
24.
25.
26.
Dziewanowski K, Drozd R, Chojnowska A, et al. Kidney transplantation among identical twins: therapeutic dilemmas. BMJ Case Rep 2011; 2011. Czyz W, Morahan JM, Ebers GC, et al. Genetic, environmental and stochastic factors in monozygotic twin discordance with a focus on epigenetic differences. BMC Med 2012; 10: 93. Bell JT, Spector TD. A twin approach to unraveling epigenetics. Trends Genet 2011; 27: 116. Solomon BD, Hadley DW, Pineda-Alvarez DE, et al. Incidental medical information in whole-exome sequencing. Pediatrics 2012; 129: e1605. Zwijnenburg PJ, Meijers-Heijboer H, Boomsma DI. Identical but not the same: the value of discordant monozygotic twins in genetic research. Am J Med Genet B Neuropsychiatr Genet 2010; 153B: 1134. Touma M, Joshi M, Connolly MC, et al. Whole genome sequencing identifies SCN2A mutation in monozygotic twins with Ohtahara syndrome and unique neuropathologic findings. Epilepsia 2013; 54: e81. Bruder CE, Piotrowski A, Gijsbers AA, et al. Phenotypically concordant and discordant monozygotic twins display different DNA copynumber-variation profiles. Am J Hum Genet 2008; 82: 763. West PM, Love DR, Stapleton PM, et al. Paternal uniparental disomy in monozygotic twins discordant for hemihypertrophy. J Med Genet 2003; 40: 223. Fraga MF, Ballestar E, Paz MF, et al. Epigenetic differences arise during the lifetime of monozygotic twins. Proc Natl Acad Sci U S A 2005; 102: 10604. Talens RP, Christensen K, Putter H, et al. Epigenetic variation during the adult lifespan: cross-sectional and longitudinal data on monozygotic twin pairs. Aging Cell 2012; 11: 694. Gordon L, Joo JH, Andronikos R, et al. Expression discordance of monozygotic twins at birth: effect of intrauterine environment and a possible mechanism for fetal programming. Epigenetics 2011; 6: 579. Forsberg LA, Absher D, Dumanski JP. Non-heritable genetics of human disease: spotlight on post-zygotic genetic variation acquired during lifetime. J Med Genet 2013; 50: 1. Khosrotehrani K, Johnson KL, Cha DH, et al. Transfer of fetal cells with multilineage potential to maternal tissue. JAMA 2004; 292: 75. Bianchi DW, Zickwolf GK, Weil GJ, et al. Male fetal progenitor cells persist in maternal blood for as long as 27 years postpartum. Proc Natl Acad Sci U S A 1996; 93: 705. Nelson JL. The otherness of self: microchimerism in health and disease. Trends Immunol 2012; 33: 421. Stevens AM, McDonnell WM, Mullarkey ME, et al. Liver biopsies from human females contain male hepatocytes in the absence of transplantation. Lab Invest 2004; 84: 1603. Bayes-Genis A, Bellosillo B, de la Calle O, et al. Identification of male cardiomyocytes of extracardiac origin in the hearts of women with male progeny: male fetal cell microchimerism of the heart. J Heart Lung Transplant 2005; 24: 2179. Lee TH, Donegan E, Slichter S, et al. Transient increase in circulating donor leukocytes after allogeneic transfusions in immunocompetent recipients compatible with donor cell proliferation. Blood 1995; 85: 1207. Starzl TE, Demetris AJ, Trucco M, et al. Chimerism after liver transplantation for type IV glycogen storage disease and type 1 Gaucher’s disease. N Engl J Med 1993; 328: 745. O’Huallachain M, Karczewski KJ, Weissman SM, et al. Extensive genetic variation in somatic human tissues. Proc Natl Acad Sci U S A 2012; 109: 18018. Moayyeri A, Hammond CJ, Hart DJ, et al. The UK Adult Twin Registry (TwinsUK Resource). Twin Res Hum Genet 2013; 16: 144.
27. 28. 29. 30.
31. 32. 33. 34. 35. 36. 37. 38. 39. 40. 41. 42. 43. 44. 45. 46. 47. 48.
489
Machin G. Non-identical monozygotic twins, intermediate twin types, zygosity testing, and the non-random nature of monozygotic twinning: a review. Am J Med Genet C Semin Med Genet 2009; 151C: 110. Montgomery GW, Zhu G, Hottenga JJ, et al. HLA and genomewide allele sharing in dizygotic twins. Am J Hum Genet 2006; 79: 1052. Baranzini SE, Mudge J, van Velkinburgh JC, et al. Genome, epigenome and RNA sequences of monozygotic twins discordant for multiple sclerosis. Nature 2010; 464: 1351. Breckpot J, Thienpont B, Gewillig M, et al. Differences in copy number variation between discordant monozygotic twins as a model for exploring chromosomal mosaicism in congenital heart defects. Mol Syndromol 2012; 2: 81. Worthington J, Silman AJ. Genetic control of autoimmunity, lessons from twin studies. Clin Exp Immunol 1995; 101: 390. Gregersen PK. Discordance for autoimmunity in monozygotic twins. Are ‘‘identical’’ twins really identical? Arthritis Rheum 1993; 36: 1185. Zielinski A, Nazarewski S, Bogetti D, et al. Simultaneous pancreaskidney transplant from living related donor: a single-center experience. Transplantation 2003; 76: 547. Benedetti E, Dunn T, Massad MG, et al. Successful living related simultaneous pancreas-kidney transplant between identical twins. Transplantation 1999; 67: 915. Liu LU, Schiano TD, Min AD, et al. Syngeneic living-donor liver transplantation without the use of immunosuppression. Gastroenterology 2002; 123: 1341. Yoshizawa A, Takada Y, Fujimoto Y, et al. Liver transplantation from an identical twin without immunosuppression, with early recurrence of hepatitis C. Am J Transplant 2006; 6: 2812. Sugawara Y, Ohtsuka H, Kaneko J, et al. Successful treatment of hepatitis C virus after liver transplantation from an identical twin. Transplantation 2002; 73: 1850. Schena S, Testa G, Setty S, et al. Successful identical-twin living donor small bowel transplant for necrotizing enterovasculitis secondary to Churg-Strauss syndrome. Transpl Int 2006; 19: 594. Berney T, Genton L, Buhler LH, et al. Five-year follow-up after pediatric living related small bowel transplantation between two monozygotic twins. Transplant Proc 2004; 36: 316. Genton L, Raguso CA, Berney T, et al. Four year nutritional follow up after living related small bowel transplantation between monozygotic twins. Gut 2003; 52: 659. Silber SJ, DeRosa M, Pineda J, et al. A series of monozygotic twins discordant for ovarian failure: ovary transplantation (cortical versus microvascular) and cryopreservation. Hum Reprod 2008; 23: 1531. Hazani R, Coots BK, Buntic RF, et al. Bilateral breast reconstruction: the simultaneous use of autogenous tissue and identical twin isograft. Ann Plast Surg 2009; 63: 496. Cheigh JS, Chami J, Stenzel KH, et al. Renal transplantation between HLA identical siblings. Comparison with transplants from HLA semiidentical related donors. N Engl J Med 1977; 296: 1030. Matzinger P. The danger model: a renewed sense of self. Science 2002; 296: 301. Nickerson PW, Rush DN. Antibodies beyond HLA. Am J Transplant 2013; 13: 831. Bramstedt KA. Living donor transplantation between twins: guidance for Donor Advocate Teams. Clin Transplant 2007; 21: 144. de Mattos AM, Bennett WM, Barry JM, et al. HLA-identical sibling renal transplantationVa 21-yr single-center experience. Clin Transplant 1999; 13: 158. Shimmura H, Tanabe K, Ishida H, et al. Long-term results of living kidney transplantation from HLA-identical sibling donors under calcineurin inhibitor immunosuppression. Int J Urol 2006; 13: 502.
Copyright © 2014 Lippincott Williams & Wilkins. Unauthorized reproduction of this article is prohibited.
Transplantation Between Monozygotic Twins: How Identical Are They? Elizabeth Day,1,5 Patrick K. Kearns,1 Craig J. Taylor,2,3 and J. Andrew Bradley 3,4 Abstract. Advances in developmental biology have shown that monozygous twins may not be as phenotypically identical as once believed, and the mechanisms responsible for such differences are now becoming clearer. Whether such phenotypic differences are capable of triggering graft rejection of an organ transplanted between identical twins remains unknown but the risks seem low, and long-term transplant outcome is excellent. Available evidence to guide immunosuppressive therapy in this setting is limited but a prudent approach would include the use of steroids together with a calcineurin inhibitor after transplantation. However, once the inevitable inflammatory response associated with transplant surgery has resolved, cautious reduction and eventually withdrawal of immunosuppression should be possible. Keywords: Twin, Transplantation, Genetic. (Transplantation 2014;98: 485Y489)
he first successful renal transplant was carried out between ‘‘identical’’ twins almost six decades ago and since that landmark case over 200 kidney transplants between identical twins have been recorded in the United States (1, 2). The early cases of renal transplantation between identical twins were carried out without immunosuppression by necessity because at the time safe and effective immunosuppression was not available. In subsequent years, successful transplantation of kidneys between identical twins has continued with the use of minimal or no immunosuppression (3Y5).
T
The authors declare no funding or conflicts of interest. 1 Department of Surgery, Southern General Hospital, Glasgow, United Kingdom. 2 Histocompatibility and Immunogenetics Laboratory, Cambridge University Teaching Hospitals NHS Foundation Trust, Addenbrooke’s Hospital, Cambridge, United Kingdom. 3 NIHR Biomedical Research Centre, Cambridge, United Kingdom. 4 Department of Surgery, University of Cambridge Clinical School, Addenbrooke’s Hospital, Cambridge, United Kingdom. 5 Address correspondence to: Elizabeth Day, Ph.D., Department of Surgery, Southern General Hospital, Glasgow, Lanarkshire, G51 4TF, United Kingdom. E-mail: [email protected] E.D. conceived and drafted, and helped revise the review. P.K.K. contributed to first and subsequent drafts of the review. C.J.T. participated in editing the first draft and subsequent drafts of the review. J.A.B. contributed to the first and subsequent drafts of the review. All the authors have approved the article for publication. Supplemental digital content (SDC) is available for this article. Direct URL citations appear in the printed text, and links to the digital files are provided in the HTML text of this article on the journal’s Web site (www.transplantjournal.com). Received 17 December 2013. Revision requested 24 February 2014. Accepted 1 May 2014. Copyright * 2014 by Lippincott Williams & Wilkins ISSN: 0041-1337/14/9805-485 DOI: 10.1097/TP.0000000000000274
Transplantation
& Volume 98, Number 5, September15, 2014
Interestingly, however, our current understanding of the molecular basis of development and the in utero environment suggests that monozygous (MZ) twins may not be as identical as previously believed. The clinical relevance of these differences in organ transplantation is unclear, and there is little study data pertaining to necessity of immunosuppressive therapy in identical twin transplantation (6). We provide a brief review of the current status of MZ twin transplantation and examine the practical and theoretical implications of current genetic, epigenetic, and immunologic research, with a particular focus on renal transplantation.
HOW IDENTICAL ARE IDENTICAL TWINS? MZ twins arise from a single ovum fertilized by a single sperm. Traditionally they are considered genetically identical but phenotypically MZ twins do show differences. Historically, these phenotypic differences have been attributed to environmental influences, including effects on epigenetic reprogramming (7). The interaction between genes and modulating environmental stimuli begins early in utero and lifestyle differences give rise to genetic variation that increases with age (epigenetic drift). Genome-wide and single locus DNA methylation patterns leads to epigenetic variability in gene promoters that cause differences in transcriptional regulation and complex phenotypes (8). Furthermore, in the last decade, a number of studies have shown that MZ twins can and do differ genetically, with changes affecting single base pairs, and rarely at larger genomic regions and whole chromosomes (9Y12). This phenomenon is related to the concept of somatic variation, whereby the genetic and epigenetic contents of cells in a single individual differ. Somatic variation and MZ twin discordance arises through three main mechanisms: somatic mosaicism, chimerism, and www.transplantjournal.com
Copyright © 2014 Lippincott Williams & Wilkins. Unauthorized reproduction of this article is prohibited.
485
486
www.transplantjournal.com
epigenetic alterations accumulated throughout life (epigenetic drift) (Fig. 1). Somatic mosaicism is not a new concept and is vital for the normal functioning of the immune system with the controlled reshuffling of immunoglobulin and T-cell receptor genes; MZ twins are confronted with different foreign antigens and hence their T-cell reservoirs will vary. However, the presence and prevalence of somatic mosaicism in other tissues is a more recent observation. Somatic mosaics originate
Transplantation
& Volume 98, Number 5, September15, 2014
from a single zygote (the earliest stage of an embryo development) but contain cells with differing genetic content. Somatic mosaics can arise through a number of mechanisms, including mutations and structural changes occurring after the zygote stage, heteroplasmy (unequal division of mitochondrial DNA within the cellular cytoplasm) and uniparental disomy (where both copies of a chromosome or genomic region are inherited from a single parent) (13). In addition, development and maintenance of any single cell within an
FIGURE 1. Mechanisms of somatic variation. A, somatic mosaicism. Genetic changes occur spontaneously in the early stages of embryo development. These changes are propagated during the development of the fetus and result in a genetic mosaicVtissues contain cells with varying genetic content (gray shades). B, chimerism. Novel genetic material is introduced from an exogenous source. This source may be maternal or from a synchronous fetus. Cells or DNA fragments are believed to pass into the fetal circulation across the placenta or through the amniotic fluid. These cells or DNA must then integrate and propagate to contribute to the chimera. C, epigenetic drift. Epigenetic changes occur over time and, as with somatic mosaicism and chimerism, are propagated to result in a mix of expression patterns within a tissue.
Copyright © 2014 Lippincott Williams & Wilkins. Unauthorized reproduction of this article is prohibited.
Day et al.
* 2014 Lippincott Williams & Wilkins
TABLE 1.
487
Case reports of solid organ and composite tissue transplants between identical twins
Kidney A single case after graft failure from an earlier transplant. Three cases. Primary renal disease recurred in all cases after discontinuation of immunosuppression. Case series of simultaneous pancreas-kidney transplant including one identical twin pair. A single case of a simultaneous pancreas-kidney transplant in an insulin dependent diabetic transplanted after the recognition of renal failure. Liver Two cases requiring transplantation for hepatitis C related cirrhosis. Two cases requiring transplantation for hepatocellular carcinoma and a carcinoid tumor. A single case requiring transplantation for hepatitis C related cirrhosis. Small Bowel A single case of transplantation after gastrointestinal necrotizing vasculitis. A single case of transplantation of terminal ileum after resection for a mid-gut volvulus. Skin graft testing was performed before transplantation. A single case with a 4-yr follow up. Other Ovarian. Case seriesVseven ovarian transplants, five pregnancies at the time of follow up. Composite Tissue: A single case of a deep inferior epigastric flap donated for breast reconstruction.
Immunosuppression
Reference
Oral steroids and mofetil mycophenolate. Discontinued at 12, 15, and 91 mo after transplantation. Maintained on low-dose cyclosporine.
(6) (4)
Azathioprine for 1 mo and maintenance on low dose cyclosporine.
(34)
V
(35)
V
(36)
(33)
Steroid therapy for 1 mo
(37)
Cyclophosphamide initially and maintenance on oral steroids. V
(38) (39)
V
(40)
V
(41)
V
(42)
Selected case reports published over the last 15 years (1998Y2014) of solid organ and composite tissue transplantation between identical twins. The second column indicates whether immunosuppression was used, a (V) indicates that no immunosuppression was used.
organism is stochastic; genetic and epigenetic modifications will continue throughout life in response to the environment and ageing. Indeed, studies of MZ twins have shown that both epigenetic and genetic changes occur in response to the in utero environment and increase with age (14Y17). A chimera originates from the mixing of cells from individuals originating from two distinct zygotes. Cells must mix, implant and survive in the new organism. This may occur ‘‘naturally’’ in utero, maternal cells entering the fetal circulation, and conversely during pregnancy, whereby fetal cells enter the maternal circulation (18, 19). It may also take place between embryos in multiple pregnancies (20). The extent of this phenomenon is unknown but has been seen in hematopoietic tissue as well as hepatocytes, myocytes and islet A cells (20Y22). Indeed, in one recent review, the author suggests ‘‘that a shift is needed from the conventional paradigm of ‘self versus other’ to a view of the normal ‘self ’ as constitutively chimeric’’ (20). Iatrogenic chimerism occurs transiently after blood transfusion or more permanently after tissue and organ transplantation (23, 24). The full extent of somatic mosaicism or chimerism in the MZ twin population has not been studied: case report data tends to focus on overtly discordant twins. Indeed, it is technically difficult to estimate the prevalence of such
variation in the larger non-twin population, and recent studies have only begun to assess the level of genomic and epigenomic diversity within a single individual (17, 25). However, with the construction of large databases containing phenotypic and genetic information from twins, our understanding of normal twin variation is set to expand (15). The twin model is particularly powerful method for assessing somatic variation, and as such, the understanding we gain from studying twin-twin variation may well prove useful in the future to the transplant team (26). Zygosity Testing A comprehensive review of twin zygosity testing is beyond the scope of this article, but it remains an important consideration in the context of organ transplantation between twins. In general monochorionicty is a good screen for monozygosity (27). Standard human leukocyte antigen (HLA) typing is not sufficient alone to determine monozygosity because 25% of dizygotic twins will match at all HLA loci (28). More extensive haplotyping, such as that used in bone marrow transplantation, may add strength to choronicity, as would analysis of additional genetic markers, such as short tandem repeats, used in DNA fingerprinting. However, genetic markers, as with any other genomic region, may
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be affected by somatic variation, and therefore any anomalies detected by genetic testing must be considered with this in mind. The complexity of zygosity testing arises from many of the concepts that have been discussed in the preceding paragraphs; a review by Machin (27) offers a thorough discussion of the subject. In the context of organ and tissue transplantation, there is perhaps scope for more ‘‘functional’’ assays, akin to skin grafting seen in the early transplantations (1). Of course, choosing an appropriate substrate for these investigations would require substantial research, and the use of renal tissue would pose a practical challenge.
IMPLICATIONS OF MZ TWIN DISCORDANCE IN TRANSPLANTATION Clinically, MZ twin discordance has been described for single gene disorders, such as hemophilia B and Duchenne Muscular Dystrophy, as well as in complex conditions, such as multiple sclerosis and congenital heart disease (10, 29, 30). There is also a high prevalence of discordance for autoimmune conditions, such as systemic lupus erythematosus and rheumatoid arthritis: somatic and epigenetic variation between MZ twins identified by next generation (DNA) sequencing technologies may hint at potential immunologic discordance between MZ twins and hence have implications for transplantation (31, 32). Although the mechanisms through which this discordance arises are becoming apparent, translating this information into clinical decisions is a challenge. In the context of MZ twin transplantation, it is the requirement and degree of immunosuppression necessary that remains an uncertainty.
IMMUNOSUPPRESSION IN MZ TRANSPLANTATION Immunosuppressive regimens must balance the risks of immunosuppression, namely, infection, malignancy, and other specific side effects of agents, with the benefits of preventing graft rejection and disease recurrence. To ascertain the evidence base for immunosuppression in transplants performed between identical twins, we searched the literature to identify studies and reports pertaining to MZ twin organ transplantation (Appendix SI, SDC, http://links.lww.com/TP/B17). The most comprehensive analysis of MZ twin transplant cases was compiled by Krishnan and colleagues who identified close to 200 MZ twin renal transplants in the United States between 1987 and 2006. They reported that 71% of cases were receiving immunosuppressive therapy at the time of discharge from hospital (61% received steroids and 31% calcineurin inhibitors), falling to 34% 1 year and 33% 3 years after transplantation. At 1 year, 21% of recipients were still on calcineurin inhibitors, and 27% were still on steroids. Interestingly, graft function was superior in recipients who were not maintained on immunosuppression. There are limited data available pertaining to rejection rates in MZ twin transplantation to provide evidence-based guidance on the use of immunosuppression. Kessaris and colleagues (5), who identified 132 living-donor identical twin transplants that took place between 1988 and 2004 in the United Kingdom and United States, recorded similar observations to those of Krishnan et al: a large proportion of the
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patients remained on immunosuppression and there was no significant difference in graft survival between those on immunosuppression and those not. The findings from these reports suggest that renal transplantation between MZ twins can be performed safely without long-term immunosuppression. Successful living-donor liver and small bowel transplants have also been carried out without immunosuppression in MZ twins (Table 1) (4, 6, 33Y42).
DISCUSSION AND CONCLUSION It has long been observed that graft rejection is not solely a function of immune homology (43). Trauma, ischemia and reperfusion during organ transplantation stimulates cytokine release, activating immune responses and contributing to graft rejection (44). The presence of autoantibodies to cryptic tissue antigens (e.g., vimentin) associated with kidney and heart allograft rejection indicate that tissue damage may be an important initiator of graft rejection regardless of genetic identity (45). For this reason, perioperative steroids are an important component of immunosuppressive regimens and one that should most likely remain in MZ twin transplantation (2). Twin and HLA-identical sibling transplantation also carries additional considerations, not related to immunosuppression, such as distinct emotional consequences and disease recurrence: one of the most common reasons for graft loss in HLA-identical sibling transplants (46Y48). When planning renal transplantation between apparent MZ twins, zygosity testing using multilocus probes should be carried out to confirm twins are indeed monozygotic. Advances in developmental biology have clearly shown that MZ twins may not be as identical as once believed, and the mechanisms responsible for such differences are now becoming clearer. To what extent such differences are capable of triggering graft rejection remains unknown, but the risks seem low, and long-term transplant outcome is excellent. Available evidence to guide immunosuppressive is limited but a prudent approach would include the use of steroids together with a calcineurin inhibitor after transplantation. However, once the inflammatory response associated with transplant surgery has resolved, cautious reduction and eventual withdrawal of immunosuppression should be possible, thereby avoiding the long-term side effects of immunosuppression, including the detrimental effects of calcineurin inhibitors on graft function. REFERENCES 1.
2. 3.
4. 5.
Merrill JP, Murray JE, Harrison JH, et al. Successful homotransplantation of the human kidney between identical twins. J Am Med Assoc 1956; 160: 277. Krishnan N, Buchanan PM, Dzebisashvili N, et al. Monozygotic transplantation: concerns and opportunities. Am J Transplant 2008; 8: 2343. Gumprich M, Woeste G, Kohlhaw K, et al. Living related kidney transplantation between identical twins. Transplant Proc 2002; 34: 2205. Kim YK, Yoon HE, Kim SH, et al. Long-term follow-up of three identical twin transplant recipients without maintenance immunosuppressive therapy. Nephrology (Carlton) 2008; 13: 447. Kessaris N, Mukherjee D, Chandak P, et al. Renal transplantation in identical twins in United States and United Kingdom. Transplantation 2008; 86: 1572.
Copyright © 2014 Lippincott Williams & Wilkins. Unauthorized reproduction of this article is prohibited.
Day et al.
* 2014 Lippincott Williams & Wilkins
6.
7.
8. 9. 10.
11.
12.
13.
14.
15.
16.
17.
18. 19.
20. 21.
22.
23.
24.
25.
26.
Dziewanowski K, Drozd R, Chojnowska A, et al. Kidney transplantation among identical twins: therapeutic dilemmas. BMJ Case Rep 2011; 2011. Czyz W, Morahan JM, Ebers GC, et al. Genetic, environmental and stochastic factors in monozygotic twin discordance with a focus on epigenetic differences. BMC Med 2012; 10: 93. Bell JT, Spector TD. A twin approach to unraveling epigenetics. Trends Genet 2011; 27: 116. Solomon BD, Hadley DW, Pineda-Alvarez DE, et al. Incidental medical information in whole-exome sequencing. Pediatrics 2012; 129: e1605. Zwijnenburg PJ, Meijers-Heijboer H, Boomsma DI. Identical but not the same: the value of discordant monozygotic twins in genetic research. Am J Med Genet B Neuropsychiatr Genet 2010; 153B: 1134. Touma M, Joshi M, Connolly MC, et al. Whole genome sequencing identifies SCN2A mutation in monozygotic twins with Ohtahara syndrome and unique neuropathologic findings. Epilepsia 2013; 54: e81. Bruder CE, Piotrowski A, Gijsbers AA, et al. Phenotypically concordant and discordant monozygotic twins display different DNA copynumber-variation profiles. Am J Hum Genet 2008; 82: 763. West PM, Love DR, Stapleton PM, et al. Paternal uniparental disomy in monozygotic twins discordant for hemihypertrophy. J Med Genet 2003; 40: 223. Fraga MF, Ballestar E, Paz MF, et al. Epigenetic differences arise during the lifetime of monozygotic twins. Proc Natl Acad Sci U S A 2005; 102: 10604. Talens RP, Christensen K, Putter H, et al. Epigenetic variation during the adult lifespan: cross-sectional and longitudinal data on monozygotic twin pairs. Aging Cell 2012; 11: 694. Gordon L, Joo JH, Andronikos R, et al. Expression discordance of monozygotic twins at birth: effect of intrauterine environment and a possible mechanism for fetal programming. Epigenetics 2011; 6: 579. Forsberg LA, Absher D, Dumanski JP. Non-heritable genetics of human disease: spotlight on post-zygotic genetic variation acquired during lifetime. J Med Genet 2013; 50: 1. Khosrotehrani K, Johnson KL, Cha DH, et al. Transfer of fetal cells with multilineage potential to maternal tissue. JAMA 2004; 292: 75. Bianchi DW, Zickwolf GK, Weil GJ, et al. Male fetal progenitor cells persist in maternal blood for as long as 27 years postpartum. Proc Natl Acad Sci U S A 1996; 93: 705. Nelson JL. The otherness of self: microchimerism in health and disease. Trends Immunol 2012; 33: 421. Stevens AM, McDonnell WM, Mullarkey ME, et al. Liver biopsies from human females contain male hepatocytes in the absence of transplantation. Lab Invest 2004; 84: 1603. Bayes-Genis A, Bellosillo B, de la Calle O, et al. Identification of male cardiomyocytes of extracardiac origin in the hearts of women with male progeny: male fetal cell microchimerism of the heart. J Heart Lung Transplant 2005; 24: 2179. Lee TH, Donegan E, Slichter S, et al. Transient increase in circulating donor leukocytes after allogeneic transfusions in immunocompetent recipients compatible with donor cell proliferation. Blood 1995; 85: 1207. Starzl TE, Demetris AJ, Trucco M, et al. Chimerism after liver transplantation for type IV glycogen storage disease and type 1 Gaucher’s disease. N Engl J Med 1993; 328: 745. O’Huallachain M, Karczewski KJ, Weissman SM, et al. Extensive genetic variation in somatic human tissues. Proc Natl Acad Sci U S A 2012; 109: 18018. Moayyeri A, Hammond CJ, Hart DJ, et al. The UK Adult Twin Registry (TwinsUK Resource). Twin Res Hum Genet 2013; 16: 144.
27. 28. 29. 30.
31. 32. 33. 34. 35. 36. 37. 38. 39. 40. 41. 42. 43. 44. 45. 46. 47. 48.
489
Machin G. Non-identical monozygotic twins, intermediate twin types, zygosity testing, and the non-random nature of monozygotic twinning: a review. Am J Med Genet C Semin Med Genet 2009; 151C: 110. Montgomery GW, Zhu G, Hottenga JJ, et al. HLA and genomewide allele sharing in dizygotic twins. Am J Hum Genet 2006; 79: 1052. Baranzini SE, Mudge J, van Velkinburgh JC, et al. Genome, epigenome and RNA sequences of monozygotic twins discordant for multiple sclerosis. Nature 2010; 464: 1351. Breckpot J, Thienpont B, Gewillig M, et al. Differences in copy number variation between discordant monozygotic twins as a model for exploring chromosomal mosaicism in congenital heart defects. Mol Syndromol 2012; 2: 81. Worthington J, Silman AJ. Genetic control of autoimmunity, lessons from twin studies. Clin Exp Immunol 1995; 101: 390. Gregersen PK. Discordance for autoimmunity in monozygotic twins. Are ‘‘identical’’ twins really identical? Arthritis Rheum 1993; 36: 1185. Zielinski A, Nazarewski S, Bogetti D, et al. Simultaneous pancreaskidney transplant from living related donor: a single-center experience. Transplantation 2003; 76: 547. Benedetti E, Dunn T, Massad MG, et al. Successful living related simultaneous pancreas-kidney transplant between identical twins. Transplantation 1999; 67: 915. Liu LU, Schiano TD, Min AD, et al. Syngeneic living-donor liver transplantation without the use of immunosuppression. Gastroenterology 2002; 123: 1341. Yoshizawa A, Takada Y, Fujimoto Y, et al. Liver transplantation from an identical twin without immunosuppression, with early recurrence of hepatitis C. Am J Transplant 2006; 6: 2812. Sugawara Y, Ohtsuka H, Kaneko J, et al. Successful treatment of hepatitis C virus after liver transplantation from an identical twin. Transplantation 2002; 73: 1850. Schena S, Testa G, Setty S, et al. Successful identical-twin living donor small bowel transplant for necrotizing enterovasculitis secondary to Churg-Strauss syndrome. Transpl Int 2006; 19: 594. Berney T, Genton L, Buhler LH, et al. Five-year follow-up after pediatric living related small bowel transplantation between two monozygotic twins. Transplant Proc 2004; 36: 316. Genton L, Raguso CA, Berney T, et al. Four year nutritional follow up after living related small bowel transplantation between monozygotic twins. Gut 2003; 52: 659. Silber SJ, DeRosa M, Pineda J, et al. A series of monozygotic twins discordant for ovarian failure: ovary transplantation (cortical versus microvascular) and cryopreservation. Hum Reprod 2008; 23: 1531. Hazani R, Coots BK, Buntic RF, et al. Bilateral breast reconstruction: the simultaneous use of autogenous tissue and identical twin isograft. Ann Plast Surg 2009; 63: 496. Cheigh JS, Chami J, Stenzel KH, et al. Renal transplantation between HLA identical siblings. Comparison with transplants from HLA semiidentical related donors. N Engl J Med 1977; 296: 1030. Matzinger P. The danger model: a renewed sense of self. Science 2002; 296: 301. Nickerson PW, Rush DN. Antibodies beyond HLA. Am J Transplant 2013; 13: 831. Bramstedt KA. Living donor transplantation between twins: guidance for Donor Advocate Teams. Clin Transplant 2007; 21: 144. de Mattos AM, Bennett WM, Barry JM, et al. HLA-identical sibling renal transplantationVa 21-yr single-center experience. Clin Transplant 1999; 13: 158. Shimmura H, Tanabe K, Ishida H, et al. Long-term results of living kidney transplantation from HLA-identical sibling donors under calcineurin inhibitor immunosuppression. Int J Urol 2006; 13: 502.
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