Top

Published in:

01-03-2019 | Original Article

Donor-derived hepatocytes in human hematopoietic cell transplant recipients: evidence of fusion

Authors: David Myerson, Rachael K. Parkin

Published in: Virchows Archiv | Issue 3/2019

Abstract

Reconstitution of hepatocytes by hematopoietic stem cells—a phenomenon which occurs in rodents under highly selective conditions—results from infrequent fusion between incoming myelomonocytes and host hepatocytes, with subsequent proliferation. Human hematopoietic stem cell transplant recipients have been little studied, with some support for transdifferentiation (direct differentiation). We studied routinely obtained autopsy liver tissue of four female hematopoietic cell transplant recipients with male donors, using a highly specific conjoint immunohistochemistry in situ hybridization light microscopic technique. Hepatocyte nuclei were identified by cytokeratin (Cam5.2) staining and evaluated for X and Y chromosome content. Over 1.6 million hepatocytes were assessed for rare instances of donor origin, revealing a Y chromosome in 67. Mixed tetraploids (XXXY) and their nuclear truncation products (XXY, XY, Y) were directly demonstrated, with no detection of the male tetraploids (XXYY) that may result from transdifferentiation with subsequent tetraploidization, nor their unique truncation products (XYY, YY), implicating fusion as the mechanism. To determine whether it is the sole mechanism, we modeled the chromosome distribution based on the same probability of detection of each X chromosome, deriving parameters of sensitivity and female tetraploidy by best fit. We then hypothesized that the distribution of Y chromosome–containing cells could be predicted by a similar model. After modification to account for “clumpy” Y chromosomes, the observed results were in accord with the predicted results (p = 0.6). These results suggest that all the Y-containing cells, including apparent XY cells, derive from mixed tetraploids, consistent with fusion as the sole mechanism.

Available only for authorised users

Shafritz DA, Oertel M (2011) Model systems and experimental conditions that lead to effective repopulation of the liver by transplanted cells. Int J Biochem Cell Biol 43:198–213. https://doi.org/10.1016/j.biocel.2010.01.013 CrossRefPubMed

Fausto N (2004) Liver regeneration and repair: hepatocytes, progenitor cells, and stem cells. Hepatology 39:1477–1487. https://doi.org/10.1002/hep.20214 CrossRefPubMed

Kallis YN, Alison MR, Forbes SJ (2007) Bone marrow stem cells and liver disease. Gut 56:716–724. https://doi.org/10.1136/gut.2006.098442 CrossRefPubMedPubMedCentral

Lagasse E, Connors H, Al-Dhalimy M et al (2000) Purified hematopoietic stem cells can differentiate into hepatocytes in vivo. Nat Med 6:1229–1234CrossRefPubMed

Wang X, Willenbring H, Akkari Y, Torimaru Y, Foster M, al-Dhalimy M, Lagasse E, Finegold M, Olson S, Grompe M (2003) Cell fusion is the principal source of bone-marrow-derived hepatocytes. Nature 422:897–901CrossRefPubMed

Vassilopoulos G, Wang PR, Russell DW (2003) Transplanted bone marrow regenerates liver by cell fusion. Nature 422:901–904CrossRefPubMed

Resaz R, Emionite L, Vanni C, Astigiano S, Puppo M, Lavieri R, Segalerba D, Pezzolo A, Bosco MC, Oberto A, Eva C, Chou JY, Varesio L, Barbieri O, Eva A (2011) Treatment of newborn G6pc^−/− mice with bone marrow-derived myelomonocytes induces liver repair. J Hepatol 55:1263–1271. https://doi.org/10.1016/j.jhep.2011.02.033 CrossRefPubMedPubMedCentral

Willenbring H, Bailey AS, Foster M, Akkari Y, Dorrell C, Olson S, Finegold M, Fleming WH, Grompe M (2004) Myelomonocytic cells are sufficient for therapeutic cell fusion in liver. Nat Med 10:744–748. https://doi.org/10.1038/nm1062 CrossRefPubMed

Camargo FD, Finegold M, Goodell MA (2004) Hematopoietic myelomonocytic cells are the major source of hepatocyte fusion partners. J Clin Invest 113:1266–1270. https://doi.org/10.1172/JCI21301 CrossRefPubMedPubMedCentral

10.

Duncan AW, Hickey RD, Paulk NK, Culberson AJ, Olson SB, Finegold MJ, Grompe M (2009) Ploidy reductions in murine fusion-derived hepatocytes. PLoS Genet 5:e1000385. https://doi.org/10.1371/journal.pgen.1000385 CrossRefPubMedPubMedCentral

11.

Thorgeirsson SS, Grisham JW (2006) Hematopoietic cells as hepatocyte stem cells: a critical review of the evidence. Hepatology 43:2–8. https://doi.org/10.1002/hep.21015 CrossRefPubMed

12.

Wagers AJ, Sherwood RI, Christensen JL, Weissman IL (2002) Little evidence for developmental plasticity of adult hematopoietic stem cells. Science 297:2256–2259CrossRefPubMed

13.

Grompe M (2005) Bone marrow-derived hepatocytes. Novartis Found Symp 265:20–27 discussion 28–34– 92–7PubMed

14.

Theise ND, Badve S, Saxena R, Henegariu O, Sell S, Crawford JM, Krause DS (2000) Derivation of hepatocytes from bone marrow cells in mice after radiation-induced myeloablation. Hepatology 31:235–240. https://doi.org/10.1002/hep.510310135 CrossRefPubMed

15.

Körbling M, Katz RL, Khanna A, Ruifrok AC, Rondon G, Albitar M, Champlin RE, Estrov Z (2002) Hepatocytes and epithelial cells of donor origin in recipients of peripheral-blood stem cells. N Engl J Med 346:738–746CrossRefPubMed

16.

Alison MR, Poulsom R, Jeffery R, Dhillon AP, Quaglia A, Jacob J, Novelli M, Prentice G, Williamson J, Wright NA (2000) Hepatocytes from non-hepatic adult stem cells. Nature 406:257CrossRefPubMed

17.

Wu T, Cieply K, Nalesnik MA, Randhawa PS, Sonzogni A, Bellamy C, Abu-Elmagd K, Michalopolous GK, Jaffe R, Kormos RL, Gridelli B, Fung JJ, Demetris AJ (2003) Minimal evidence of transdifferentiation from recipient bone marrow to parenchymal cells in regenerating and long-surviving human allografts. Am J Transplant 3:1173–1181CrossRefPubMed

18.

Lizier M, Castelli A, Montagna C, Lucchini F, Vezzoni P, Faggioli F (2018) Cell fusion in the liver, revisited. WJH 10:213–221. https://doi.org/10.4254/wjh.v10.i2.213 CrossRefPubMed

19.

Myerson D, Parkin RK, Benirschke K, Tschetter CN, Hyde SR (2006) The pathogenesis of villitis of unknown etiology: analysis with a new conjoint immunohistochemistry-in situ hybridization procedure to identify specific maternal and fetal cells. Pediatr Dev Pathol 9:257–265. https://doi.org/10.2350/08-05-0103.1 CrossRefPubMed

20.

Willard HF, Smith KD, Sutherland J (1983) Isolation and characterization of a major tandem repeat family from the human X chromosome. Nucleic Acids Res 11:2017–2033CrossRefPubMedPubMedCentral

21.

Lau YF, Dozy AM, Huang JC, Kan YW (1984) A rapid screening test for antenatal sex determination. Lancet 323:14–16. https://doi.org/10.1016/s0140-6736(84)90182-x CrossRef

22.

Kudryavtsev BN, Kudryavtseva MV, Sakuta GA, Stein GI (1993) Human hepatocyte polyploidization kinetics in the course of life cycle. Virchows Arch B Cell Pathol 64:387–393CrossRef

23.

Toyoda H, Bregerie O, Vallet A, Nalpas B, Pivert G, Brechot C, Desdouets C (2005) Changes to hepatocyte ploidy and binuclearity profiles during human chronic viral hepatitis. Gut 54:297–302. https://doi.org/10.1136/gut.2004.043893 CrossRefPubMedPubMedCentral

24.

Gentric G, Desdouets C (2014) Polyploidization in liver tissue. Am J Pathol 184:322–331. https://doi.org/10.1016/j.ajpath.2013.06.035 CrossRefPubMed

25.

Duncan AW, Hanlon Newell AE, Smith L, Wilson EM, Olson SB, Thayer MJ, Strom SC, Grompe M (2012) Frequent aneuploidy among normal human hepatocytes. Gastroenterology 142:25–28. https://doi.org/10.1053/j.gastro.2011.10.029 CrossRefPubMed

26.

Knouse KA, Wu J, Whittaker CA, Amon A (2014) Single cell sequencing reveals low levels of aneuploidy across mammalian tissues. Proc Natl Acad Sci U S A 111:13409–13414. https://doi.org/10.1073/pnas.1415287111 CrossRefPubMedPubMedCentral

27.

Stevens AM, Michael McDonnell W, Mullarkey ME, Pang JM, Leisenring W, Lee Nelson J (2004) Liver biopsies from human females contain male hepatocytes in the absence of transplantation. Lab Investig 84:1603–1609. https://doi.org/10.1038/labinvest.3700193 CrossRefPubMed

28.

Bianchi DW (2007) Fetomaternal cell trafficking: a story that begins with prenatal diagnosis and may end with stem cell therapy. J Pediatr Surg 42:12–18. https://doi.org/10.1016/j.jpedsurg.2006.09.047 CrossRefPubMed

29.

Rijnink EC, Penning ME, Wolterbeek R, Wilhelmus S, Zandbergen M, van Duinen SG, Schutte J, Bruijn JA, Bajema IM (2015) Tissue microchimerism is increased during pregnancy: a human autopsy study. Mol Hum Reprod 21:857–864. https://doi.org/10.1093/molehr/gav047 CrossRefPubMed

30.

Boddy AM, Fortunato A, Wilson Sayres M, Aktipis A (2015) Fetal microchimerism and maternal health: a review and evolutionary analysis of cooperation and conflict beyond the womb. Bioessays 37:1106–1118. https://doi.org/10.1002/bies.201500059 CrossRefPubMedPubMedCentral

31.

Guettier C, Sebagh M, Buard J, Feneux D, Ortin-Serrano M, Gigou M, Tricottet V, Reynès M, Samuel D, Féray C (2005) Male cell microchimerism in normal and diseased female livers from fetal life to adulthood. Hepatology 42:35–43. https://doi.org/10.1002/hep.20761 CrossRefPubMed

32.

Khosrotehrani K, Bianchi DW (2005) Multi-lineage potential of fetal cells in maternal tissue: a legacy in reverse. J Cell Sci 118:1559–1563. https://doi.org/10.1242/jcs.02332 CrossRefPubMed

33.

O'Donoghue K, Sultan HA, Al-Allaf FA (2008) Microchimeric fetal cells cluster at sites of tissue injury in lung decades after pregnancy. Reprod BioMed Online 16:382–390. https://doi.org/10.1016/s1472-6483(10)60600-1 CrossRefPubMed

34.

Weimann JM, Charlton CA, Brazelton TR, Hackman RC, Blau HM (2003) Contribution of transplanted bone marrow cells to Purkinje neurons in human adult brains. Proc Natl Acad Sci U S A 100:2088–2093. https://doi.org/10.1073/pnas.0337659100 CrossRefPubMedPubMedCentral

35.

Weimann JM, Johansson CB, Trejo A, Blau HM (2003) Stable reprogrammed heterokaryons form spontaneously in Purkinje neurons after bone marrow transplant. Nat Cell Biol 5:959–966. https://doi.org/10.1038/ncb1053 CrossRefPubMed

36.

Johansson CB, Youssef S, Koleckar K, Holbrook C, Doyonnas R, Corbel SY, Steinman L, Rossi FMV, Blau HM (2008) Extensive fusion of haematopoietic cells with Purkinje neurons in response to chronic inflammation. Nat Cell Biol 10:575–583. https://doi.org/10.1038/ncb1720 CrossRefPubMedPubMedCentral

Title: Donor-derived hepatocytes in human hematopoietic cell transplant recipients: evidence of fusion
Authors: David Myerson
Rachael K. Parkin
Publication date: 01-03-2019
Publisher: Springer Berlin Heidelberg
Published in: Virchows Archiv / Issue 3/2019
Print ISSN: 0945-6317
Electronic ISSN: 1432-2307
DOI: https://doi.org/10.1007/s00428-018-2497-8

At a glance: The STEP trials

Springer Medicine

Donor-derived hepatocytes in human hematopoietic cell transplant recipients: evidence of fusion

Abstract

At a glance: The STEP trials

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 3/2019

Clinicopathologic analysis and subclassification of benign lipomatous lesions of the colon

Histologic diagnosis and grading of esophageal acute graft-versus-host disease

Eccrine ductal and acrosyringeal metaplasia in breast carcinomas: report of eight cases

The performance of digital microscopy for primary diagnosis in human pathology: a systematic review

Metastatic breast carcinoma to the urinary bladder—a report of 11 cases including a tumor to tumor metastasis

Letter to the editor: reply to Antonio Ieni “Intratumoral HER2 heterogeneity in early gastric carcinomas: potential bias in therapeutical management”