Cell fusion between circulating bone marrow-derived cells (BMDCs) and non-hematopoietic cells is well documented in various tissues and has recently been suggested to occur in response to injury. Here we illustrate that inflammation within the intestine enhanced the level of BMDC fusion with intestinal progenitors. To identify important microenvironmental factors mediating intestinal epithelial cell fusion, we performed bone marrow transplantation into mouse models of inflammation and stimulated epithelial proliferation. Interestingly, in a non-injury model or in instances where inflammation was suppressed, an appreciable baseline level of fusion persisted. This suggests that additional mediators of cell fusion exist. A rigorous temporal analysis of early post-transplantation cellular dynamics revealed that GFP-expressing donor cells first trafficked to the intestine coincident with a striking increase in epithelial proliferation, advocating for a required fusogenic state of the host partner. Directly supporting this hypothesis, induction of augmented epithelial proliferation resulted in a significant increase in intestinal cell fusion. Here we report that intestinal inflammation and epithelial proliferation act together to promote cell fusion. While the physiologic impact of cell fusion is not yet known, the increased incidence in an inflammatory and proliferative microenvironment suggests a potential role for cell fusion in mediating the progression of intestinal inflammatory diseases and cancer.
References
[1]
Alvarez-Dolado M, Pardal R, Garcia-Verdugo JM, Fike JR, Lee HO, et al. (2003) Fusion of bone-marrow-derived cells with Purkinje neurons, cardiomyocytes and hepatocytes. Nature 425: 968–973.
[2]
Camargo FD, Finegold M, Goodell MA (2004) Hematopoietic myelomonocytic cells are the major source of hepatocyte fusion partners. J Clin Invest 113: 1266–1270.
[3]
Camargo FD, Green R, Capetanaki Y, Jackson KA, Goodell MA (2003) Single hematopoietic stem cells generate skeletal muscle through myeloid intermediates. Nat Med 9: 1520–1527.
[4]
Corbel SY, Lee A, Yi L, Duenas J, Brazelton TR, et al. (2003) Contribution of hematopoietic stem cells to skeletal muscle. Nat Med 9: 1528–1532.
[5]
Ferrari G, Cusella-De Angelis G, Coletta M, Paolucci E, Stornaiuolo A, et al. (1998) Muscle regeneration by bone marrow-derived myogenic progenitors. Science 279: 1528–1530.
[6]
Lagasse E, Connors H, Al-Dhalimy M, Reitsma M, Dohse M, et al. (2000) Purified hematopoietic stem cells can differentiate into hepatocytes in vivo. Nat Med 6: 1229–1234.
[7]
Nygren JM, Jovinge S, Breitbach M, Sawen P, Roll W, et al. (2004) Bone marrow-derived hematopoietic cells generate cardiomyocytes at a low frequency through cell fusion, but not transdifferentiation. Nat Med 10: 494–501.
[8]
Rizvi AZ, Swain JR, Davies PS, Bailey AS, Decker AD, et al. (2006) Bone marrow-derived cells fuse with normal and transformed intestinal stem cells. Proc Natl Acad Sci U S A 103: 6321–6325.
[9]
Vassilopoulos G, Wang PR, Russell DW (2003) Transplanted bone marrow regenerates liver by cell fusion. Nature 422: 901–904.
[10]
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.
[11]
Johansson CB, Youssef S, Koleckar K, Holbrook C, Doyonnas R, et al. (2008) Extensive fusion of haematopoietic cells with Purkinje neurons in response to chronic inflammation. Nat Cell Biol 10: 575–583.
[12]
Nygren JM, Liuba K, Breitbach M, Stott S, Thoren L, et al. (2008) Myeloid and lymphoid contribution to non-haematopoietic lineages through irradiation-induced heterotypic cell fusion. Nat Cell Biol 10: 584–592.
[13]
Karin M (2008) The IkappaB kinase - a bridge between inflammation and cancer. Cell Res 18: 334–342.
[14]
Karin M, Greten FR (2005) NF-kappaB: linking inflammation and immunity to cancer development and progression. Nat Rev Immunol 5: 749–759.
[15]
Karin M (2005) Inflammation and cancer: the long reach of Ras. Nat Med 11: 20–21.
[16]
Nelson D, Ganss R (2006) Tumor growth or regression: powered by inflammation. J Leukoc Biol 80: 685–690.
[17]
Bernstein CN, Blanchard JF, Kliewer E, Wajda A (2001) Cancer risk in patients with inflammatory bowel disease: a population-based study. Cancer 91: 854–862.
[18]
Eaden JA, Abrams KR, Mayberry JF (2001) The risk of colorectal cancer in ulcerative colitis: a meta-analysis. Gut 48: 526–535.
[19]
Berg DJ, Davidson N, Kuhn R, Muller W, Menon S, et al. (1996) Enterocolitis and colon cancer in interleukin-10-deficient mice are associated with aberrant cytokine production and CD4(+) TH1-like responses. J Clin Invest 98: 1010–1020.
Rennick D, Davidson N, Berg D (1995) Interleukin-10 gene knock-out mice: a model of chronic inflammation. Clin Immunol Immunopathol 76: S174–178.
[22]
Wang X, Willenbring H, Akkari Y, Torimaru Y, Foster M, et al. (2003) Cell fusion is the principal source of bone-marrow-derived hepatocytes. Nature 422: 897–901.
[23]
Anderson DA, Wu Y, Jiang S, Zhang X, Streeter PR, et al. (2005) Donor marker infidelity in transgenic hematopoietic stem cells. Stem Cells 23: 638–643.
[24]
Nakanishi T, Kuroiwa A, Yamada S, Isotani A, Yamashita A, et al. (2002) FISH analysis of 142 EGFP transgene integration sites into the mouse genome. Genomics 80: 564–574.
[25]
Azad Khan AK, Piris J, Truelove SC (1977) An experiment to determine the active therapeutic moiety of sulphasalazine. Lancet 2: 892–895.
[26]
Dionne S, Hiscott J, D'Agata I, Duhaime A, Seidman EG (1997) Quantitative PCR analysis of TNF-alpha and IL-1 beta mRNA levels in pediatric IBD mucosal biopsies. Dig Dis Sci 42: 1557–1566.
[27]
Kokkotou E, Moss AC, Torres D, Karagiannides I, Cheifetz A, et al. (2008) Melanin-concentrating hormone as a mediator of intestinal inflammation. Proc Natl Acad Sci U S A 105: 10613–10618.
[28]
Molla M, Panes J (2007) Radiation-induced intestinal inflammation. World J Gastroenterol 13: 3043–3046.
[29]
Abkowitz JL, Robinson AE, Kale S, Long MW, Chen J (2003) Mobilization of hematopoietic stem cells during homeostasis and after cytokine exposure. Blood 102: 1249–1253.
[30]
Bunster E, Meyer RK (1933) An improved method of parabiosis. Anat Rec 57: 339–343.
[31]
Wright DE, Wagers AJ, Gulati AP, Johnson FL, Weissman IL (2001) Physiological migration of hematopoietic stem and progenitor cells. Science 294: 1933–1936.
[32]
Jurjus AR, Khoury NN, Reimund JM (2004) Animal models of inflammatory bowel disease. J Pharmacol Toxicol Methods 50: 81–92.
[33]
Nern C, Wolff I, Macas J, von Randow J, Scharenberg C, et al. (2009) Fusion of hematopoietic cells with Purkinje neurons does not lead to stable heterokaryon formation under noninvasive conditions. J Neurosci 29: 3799–3807.
[34]
Potten CS (1990) A comprehensive study of the radiobiological response of the murine (BDF1) small intestine. Int J Radiat Biol 58: 925–973.
[35]
Potten CS, Owen G, Roberts SA (1990) The temporal and spatial changes in cell proliferation within the irradiated crypts of the murine small intestine. Int J Radiat Biol 57: 185–199.
[36]
Davies PS, Dismuke AD, Powell AE, Carroll KH, Wong MH (2008) Wnt-reporter expression pattern in the mouse intestine during homeostasis. BMC Gastroenterol 8: 57.
[37]
Kovacs L, Potten CS (1973) An estimation of proliferative population size in stomach, jejunum and colon of DBA-2 mice. Cell Tissue Kinet 6: 125–134.
[38]
Sacco A, Doyonnas R, Kraft P, Vitorovic S, Blau HM (2008) Self-renewal and expansion of single transplanted muscle stem cells. Nature 456: 502–506.
[39]
Ireland H, Kemp R, Houghton C, Howard L, Clarke AR, et al. (2004) Inducible Cre-mediated control of gene expression in the murine gastrointestinal tract: effect of loss of beta-catenin. Gastroenterology 126: 1236–1246.
[40]
Shibata H, Toyama K, Shioya H, Ito M, Hirota M, et al. (1997) Rapid colorectal adenoma formation initiated by conditional targeting of the Apc gene. Science 278: 120–123.
[41]
Sansom OJ, Reed KR, Hayes AJ, Ireland H, Brinkmann H, et al. (2004) Loss of Apc in vivo immediately perturbs Wnt signaling, differentiation, and migration. Genes Dev 18: 1385–1390.
[42]
Cheng H, Merzel J, Leblond CP (1969) Renewal of Paneth cells in the small intestine of the mouse. Am J Anat 126: 507–525.
[43]
Madison BB, Dunbar L, Qiao XT, Braunstein K, Braunstein E, et al. (2002) Cis elements of the villin gene control expression in restricted domains of the vertical (crypt) and horizontal (duodenum, cecum) axes of the intestine. J Biol Chem 277: 33275–33283.
[44]
Moser AR, Pitot HC, Dove WF (1990) A dominant mutation that predisposes to multiple intestinal neoplasia in the mouse. Science 247: 322–324.
[45]
Soriano P (1999) Generalized lacZ expression with the ROSA26 Cre reporter strain. Nat Genet 21: 70–71.
[46]
Battaile KP, Bateman RL, Mortimer D, Mulcahy J, Rathbun RK, et al. (1999) In vivo selection of wild-type hematopoietic stem cells in a murine model of Fanconi anemia. Blood 94: 2151–2158.
[47]
Bailey AS, Willenbring H, Jiang S, Anderson DA, Schroeder DA, et al. (2006) Myeloid lineage progenitors give rise to vascular endothelium. Proc Natl Acad Sci U S A 103: 13156–13161.
[48]
Wong MH, Rubinfeld B, Gordon JI (1998) Effects of forced expression of an NH2-terminal truncated beta-Catenin on mouse intestinal epithelial homeostasis. J Cell Biol 141: 765–777.
[49]
Azuma H, Paulk N, Ranade A, Dorrell C, Al-Dhalimy M, et al. (2007) Robust expansion of human hepatocytes in Fah-/-/Rag2-/-/Il2rg-/- mice. Nat Biotechnol 25: 903–910.
[50]
Wong MH, Saam JR, Stappenbeck TS, Rexer CH, Gordon JI (2000) Genetic mosaic analysis based on Cre recombinase and navigated laser capture microdissection. Proc Natl Acad Sci U S A 97: 12601–12606.
