Cardiogenol C can induce Mouse Hair Bulge Progenitor Cells to Transdifferentiate into Cardiomyocyte-like Cells
Tóm tắt
Hair bulge progenitor cells (HBPCs) are multipotent stem cells derived from the bulge region of mice vibrissal hairs. The purified HBPCs express CD34, K15 and K14 surface markers. It has been reported that HBPCs could be readily induced to transdifferentiate into adipocytes and osteocytes. However, the ability of HBPCs to transdifferentiate into cardiomyocytes has not yet been investigated. The cardiomyogenic potential of HBPCs was investigated using a small cell-permeable molecule called Cardiogenol C. We established that Cardiogenol C could induce HBPCs to express transcription factors GATA4, Nkx2.5 and Tbx5, which are early specific markers for pre-cardiomyogenic cells. In prolonged cultures, the Cardiogenol C-treated HBPCs can also express muscle proteins, cardiac-specific troponin I and sarcomeric myosin heavy chain. However, we did not observe the ability of these cells to functionally contract. Hence, we called these cells cardiomyocyte-like cells rather than cardiomyocytes. We tried to remedy this deficiency by pre-treating HBPCs with Valproic acid first before exposing them to Cardiogenol C. This pretreatment inhibited, rather than improved, the effectiveness of Cardiogenol C in reprogramming the HBPCs. We used comparative proteomics to determine how Cardiogenol C worked by identifying proteins that were differentially expressed. We identified proteins that were involved in promoting cell differentiation, cardiomyocyte development and for the normal function of striated muscles. From those differentially expressed proteins, we further propose that Cardiogenol C might exert its effect by activating the Wnt signaling pathway through the suppression of Kremen1. In addition, by up-regulating the expression of chromatin remodeling proteins, SIK1 and Smarce1 would initiate cardiac differentiation. In conclusion, our CD34+/K15+ HBPCs could be induced to transdifferentiate into cardiomyocyte-like cells using a small molecule called Cardiogenol C. The process involves activation of the Wnt signaling pathway and altered expression of several key chromatin remodeling proteins. The finding is clinically significant as HBPCs offer a readily accessible and autologous source of progenitor cells for cell-based therapy of heart disease, which is one of major killers in developed countries.
Tài liệu tham khảo
Barth JH, Messenger AG: Measurement of hair growth and investigation of hair disease. In Diseases of the Hair and Scalp. 3rd edition. Edited by: Dawwber R. Oxford: Blackwell Scientific; 1997:564–579.
Cotsarelis G, Sun TT, Lavker RM: Label-retaining cells reside in the bulge area of pilosebaceous unit - implications for follicular stem cells, hair cycle, and skin carcinogenesis. Cell 1990, 61: 1329–1337. 10.1016/0092-8674(90)90696-C
Ohyama M, Terunuma A, Tock CL, Radonovich MF, Pise-Masison CA, et al.: Characterization and isolation of stem cell-enriched human hair follicle bulge cells. J Clin Invest 2006, 116: 249–260. 10.1172/JCI26043
Oshima H, Rochat A, Kedzia C, Kobayashi K, Barrandon Y: Morphogenesis and renewal of hair follicles from adult multipotent stem cells. Cell 2001, 104: 233–245. 10.1016/S0092-8674(01)00208-2
Sieber-Blum M, Grim M, Hu YF, Szeder V: Pluripotent neural crest stem cells in the adult hair follicle. Dev Dyn 2004, 231: 258–269. 10.1002/dvdy.20129
Rao MS: Multipotent and restricted precursors in the central nervous system. Anat Rec 2000,257(4):137–48. 10.1002/(SICI)1097-0185(19990815)257:4<137::AID-AR7>3.0.CO;2-Q
Richardson MK, Sieber-Blum M: Pluripotent neural crest cells in the developing skin of the quail embryo. Dev Biol 1993,157(2):348–58. 10.1006/dbio.1993.1140
Etchevers HC, Vincent C, Le Douarin NM, Couly GF: The cephalic neural crest provides pericytes and smooth muscle cells to all blood vessels of the face and forebrain. Development 2001, 128: 1059–1068.
Lee G, Kim H, Elkabetz Y, Al Shamy G, Panagiotakos G, Barberi T, Tabar V, Studer L: Isolation and directed differentiation of neural crest stem cells derived from human embryonic stem cells. Nat Biotechnol 2008,25(12):1468–75. 10.1038/nbt1365
Tomita Y, Matsumura K, Wakamatsu Y, Matsuzaki Y, Shibuya I, et al.: Cardiac neural crest cells contribute to the dormant multipotent stem cell in the mammalian heart. J Cell Biol 2005, 170: 1135–1146. 10.1083/jcb.200504061
Morley P, Whitfield JF: The differentiation inducer, dimethyl sulfoxide, transiently increases the intracellular calcium ion concentration in various cell types. J Cell Physiol 1993, 156: 219–225. 10.1002/jcp.1041560202
Skerjanc IS: Cardiac and skeletal muscle development in P19 embryonal carcinoma cells. Trends Cardiovasc Med 1999, 9: 139–143. 10.1016/S1050-1738(99)00017-1
Makino S, Fukuda K, Miyoshi S: Cardiomyocytes can be generated from marrow stromal cells in vitro. J Clin Invest 1999, 103: 697–705. 10.1172/JCI5298
Fukuda K: Reprogramming of bone marrow mesenchymal stem cells into cardiomyocytes. CR Biol 2002, 325: 1027–1038. 10.1016/S1631-0691(02)01524-X
Rangappa S, Fen C, Lee EH, et al.: Transformation of adult mesenchymal stem cells isolated from the fatty tissue into cardiomyocytes. Ann Thorac Surg 2003, 75: 775–779. 10.1016/S0003-4975(02)04568-X
Choi SC, Yoon J, Shim WJ, Ro YM, DS Lim: 5-Azacytidine induces cardiac differentiation of P19 embryonic stem cells. Exp Mol Med 2004, 36: 515–523.
Taylor SM, Jones PA: Changes in phenotype expression in embryonic and adult cells treated with 5-azacytidine. J Cell Physiol 1982, 111: 187–194. 10.1002/jcp.1041110210
Wu X, Ding S, Ding Q, Gray NS, Schultz PG: Small molecules that induce cardiomyogenesis in embryonic stem cells. J Am Chem Soc 2004, 126: 1590–1591. 10.1021/ja038950i
Tang MK, Wang CM, Shan SW, Chui YL, Ching AK, et al.: Comparative proteomic analysis reveals a function of the novel death receptor-associated protein BRE in the regulation of Prohibitin and p53 expression and proliferation. Proteomics 2006, 6: 2376–2385. 10.1002/pmic.200500603
Liu Y, Tang MK, Cai DQ, Li M, Wong WM, et al.: Cyclin I and p53 are differentially expressed during the terminal differentiation of the postnatal mouse heart. Proteomics 2007, 7: 23–32. 10.1002/pmic.200600456
Trempus CS, Morris RJ, Bortner CD, Cotsarelis G, Faircloth RS, et al.: Enrichment for living murine keratinocytes from the hair follicle bulge with the cell surface marker CD34. J Invest Dermatol 2003, 120: 501–511. 10.1046/j.1523-1747.2003.12088.x
Huangfu D, et al.: Induction of pluripotent stem cells by defined factors is greatly improved by small-molecule compounds. Nat Biotechnol 2008, 26: 795–797. 10.1038/nbt1418
Davidson G, Mao B, del Barco Barrantes I, Niehrs C: Kremen proteins interact with Dickkopf1 to regulate anteroposterior CNS patterning. Development 2002,129(24):5587–96. 10.1242/dev.00154
Mao B, Wu W, Davidson G, Marhold J, Li M, et al.: Kremen proteins are dickkopf receptors that regulate Wnt/beta-catenin signaling. Nature 2002, 417: 664–667. 10.1038/nature756
Pandur P, Läsche M, Eisenberg LM, Kühl M: Wnt-11 activation of a non-canonical Wnt signalling pathway is required for cardiogenesis. Nature 2002, 418: 636–41. 10.1038/nature00921
Hsu SC, Galceran J, Grosschedl R: Modulation of transcriptional regulation by LEF-1 in response to Wnt-1 signaling and association with beta-catenin. Mol Cell Biol 1998,18(8):4807–18.
Lin L, Cui L, Zhou W, Dufort D, Zhang X, Cai CL, Bu L, Yang L, Martin J, Kemler R, et al.: Beta-catenin directly regulates Islet1 expression in cardiovascular progenitors and is required for multiple aspects of cardiogenesis. Proc Natl Acad Sci USA 2007, 104: 9313–9318. 10.1073/pnas.0700923104
Berdeaux R, Goebel N, Banaszynski L, Takemori H, Wandless T, et al.: SIK1 is a class II HDAC kinase that promotes survival of skeletal myocytes. Nat Med 2007, 13: 597–603. 10.1038/nm1573
van der Linden AM, Nolan KM, Sengupta P: KIN-29 SIK regulates chemoreceptor gene expression via an MEF2 transcription factor and a class II HDAC. EMBO J 2007,26(2):358–70. 10.1038/sj.emboj.7601479
Shirai M, Osugi T, Koga H, Kaji Y, Takimoto E, Komuro I, Hara J, Miwa T, Yamauchi-Takihara K, Takihara Y: The Polycomb-group gene Rae28 sustains Nkx2.5/Csx expression and is essential for cardiac morphogenesis. J Clin Invest 2002,110(2):177–84.
Singer AJ, Clark RA: Cutaneous wound healing. N Engl J Med 1999, 341: 738–746. 10.1056/NEJM199909023411006
Morris RJ, Liu Y, Marles L, et al.: Capturing and profiling adult hair follicle stem cells. Nat Biotechnol 2004, 22: 411–417. 10.1038/nbt950
Greco V, Chen T, Rendl M, Schober M, Pasolli HA, Stokes N, Dela Cruz-Racelis J, Fuchs E: A two-step mechanism for stem cell activation during hair regeneration. Cell Stem Cell 2009,4(2):155–69. 10.1016/j.stem.2008.12.009
Ito M, Liu Y, Yang Z, Nguyen J, Liang F, Morris RJ, Cotsarelis G: Stem cells in the hair follicle bulge contribute to wound repair but not to homeostasis of the epidermis. Nat Med 2005,11(12):1351–4. 10.1038/nm1328
Fuchs E, Segre JA: Stem cells: a new lease on life. Cell 2000,100(1):143–55. 10.1016/S0092-8674(00)81691-8
Avilion AA, Nicolis SK, Pevny LH, Perez L, Vivian N, Lovell-Badge R: Multipotent cell lineages in early mouse development depend on SOX2 function. Genes Dev 2003, 17: 126–140. 10.1101/gad.224503
Lefebvre V, Dumitriu B, Penzo-Mendez A, Han Y, Pallavi B: Control of cell fate and differentiation by Sry-related high-mobility-group box (Sox) transcription factors. Int J Biochem Cell Biol 2007, 39: 2195–2214. 10.1016/j.biocel.2007.05.019
Takahashi K, Yamanaka S: Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 2006, 126: 663–667. 10.1016/j.cell.2006.07.024
Durocher D, Charron F, Warren R, Schwartz RJ, Nemer M: The cardiac transcription factors Nkx2–5 and GATA-4 are mutual cofactors. EMBO J 1997, 16: 5687–5696. 10.1093/emboj/16.18.5687
Akazawa H, Komuro I: Cardiac transcription factor Csx/Nkx2–5: Its role in cardiac development and diseases. Pharmacol Ther 2005, 107: 252–268. 10.1016/j.pharmthera.2005.03.005
Huangfu D, et al.: Induction of pluripotent stem cells from primary human fibroblasts with only Oct4 and Sox2. Nat Biotechnol 2008, 26: 1269–1275. 10.1038/nbt.1502
Liu JY, Peng HF, Andreadis ST: Contractile smooth muscle cells derived from hair-follicle stem cells. Cardiovasc Res 2008, 79: 24–33. 10.1093/cvr/cvn059
Nakamura T, Sano M, Zhou S, Schneider MD: A Wnt- and β-catenin-dependent pathway for mammalian cardiac myogenesis. Proc Natl Acad Sci USA 2003,100(10):5834–5839. 10.1073/pnas.0935626100
Kwon C, Arnold J, Hsiao EC, Taketo MM, Conklin BR, Srivastava D: Canonical Wnt signaling is a positive regulator of mammalian cardiac progenitors. Proc Natl Acad Sci USA 2007,104(26):10894–9. 10.1073/pnas.0704044104
Sheldahl LC, Park M, Malbon CC, Moon RT: Protein kinase C is differentially stimulated by Wnt and Frizzled homologs in a G-protein-dependent manner. Curr Biol 1999, 9: 695–698. 10.1016/S0960-9822(99)80310-8
Terami H, Hidaka K, Katsumata T, Iio A, Morisaki T: Wnt11 facilitates embryonic stem cell differentiation to Nkx2.5-positive cardiomyocytes, Biochem. Biophys. Res Commun 2004, 325: 968–975.
Koyanagi M, Haendeler J, Badorff C, Brandes RP, Hoffmann J, Pandur P, Kühl ZeiherAM, Dimmeler S: Non-canonical Wnt signaling enhances differentiation of human circulating progenitor cells to cardiomyogenic cells. J Biol Chem 2005, 280: 16838–16842. 10.1074/jbc.M500323200
Xu C, Police S, Rao N, Carpenter MK: Characterization and enrichment of cardiomyocytes derived from human embryonic stem cells. Circ Res 2002, 91: 501–508. 10.1161/01.RES.0000035254.80718.91
Gottlieb PD, Pierce SA, Sims RJ, Yamagishi H, Weihe EK, et al.: Bop encodes a muscle-restricted protein containing MYND and SET domains and is essential for cardiac differentiation and morphogenesis. Nat Genet 2002, 31: 25–32.
Chang S, McKinsey TA, Zhang CL, Richardson JA, Hill JA, Olson EN: Histone deacetylases 5 and 9 govern responsiveness of the heart to a subset of stress signals and play redundant roles in heart development. Mol Cell Biol 2004, 24: 8467–8476. 10.1128/MCB.24.19.8467-8476.2004
Lickert H, Takeuchi JK, Von Both I, Walls JR, McAuliffe F, et al.: Baf60c is essential for function of BAF chromatin remodelling complexes in heart development. Nature 2004, 432: 107–112. 10.1038/nature03071
Boyer LA, Plath K, Zeitlinger J, Brambrink T, Medeiros LA, et al.: Polycomb complexes repress developmental regulators in murine embryonic stem cells. Nature 2006, 441: 349–353. 10.1038/nature04733
Bracken AP, Dietrich N, Pasini D, Hansen KH, Helin K: Genome-wide mapping of polycomb target genes unravels their roles in cell fate transitions. Genes Dev 2006, 20: 1123–1136. 10.1101/gad.381706
Shao Z, et al.: Stabilization of chromatin structure by PRC1, a Polycomb complex. Cell 1999, 98: 37–46. 10.1016/S0092-8674(00)80604-2
Shan SW, Tang MK, Chow PH, Morato M: Induction of growth arrest and polycomb gene expression by reversine allows C2C12 cells to be reprogrammed to various differentiated cell types. Proteomics 2007, 7: 4303–4326. 10.1002/pmic.200700636
Ekholm SV, Reed SI: Regulation of G (1) cyclin-dependent kinases in the mammalian cell cycle. Curr Opin Cell Biol 2000, 12: 676–684. 10.1016/S0955-0674(00)00151-4
Ogasawara T, Katagiri M, Yamamoto A, Hoshi K, Takato T, et al.: Osteoclast differentiation by RANKL requires NF-kappaB-mediated downregulation of cyclin-dependent kinase 6 (Cdk6). J Bone Miner Res 2004, 19: 1128–1136. 10.1359/jbmr.2004.19.7.1128
Ekholm SV, Reed SI: Regulation of G(1) cyclin-dependent kinases in the mammalian cell cycle. Curr Opin Cell Biol 2000, 12: 676–684. 10.1016/S0955-0674(00)00151-4
Morissette MR, Cook SA, Foo S, McKoy G, Ashida N, et al.: Myostatin regulates cardiomyocyte growth through modulation of akt signaling. Circ Res 2006, 99: 15–24. 10.1161/01.RES.0000231290.45676.d4