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Signaling through the TRAIL receptor DR5/FADD pathway plays a role in the apoptosis associated with skeletal myoblast differentiation

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Abstract

Apoptosis rather than differentiation is a physiological process during myogenesis and muscle regeneration. When cultured myoblasts were induced to differentiate, we detected an increase in caspase 8 activity. Pharmacological inhibition of caspase 8 activity decreased apoptosis. Expression of a dominant-negative mutant of the adapter protein FADD also abrogated apoptosis, implicating a death ligand pathway. Treatment with TRAIL, but not Fas, induced apoptosis in these myoblasts. Accordingly, treatment with a soluble TRAIL decoy receptor or expression of a dominant-negative mutant of the TRAIL receptor DR5 abrogated apoptosis. While TRAIL expression levels remained unaltered in apoptotic myoblasts, DR5 expression levels increased. Finally, we also detected a reduction in FLIP, a death-receptor effector protein and caspase 8 competitive inhibitor, to undetectable levels in apoptotic myoblasts. Thus, our data demonstrate an important role for the TRAIL/DR5/FADD/caspase 8 pathway in the apoptosis associated with skeletal myoblast differentiation. Identifying the functional apoptotic pathways in skeletal myoblasts may prove useful in minimizing the myoblast apoptosis that contributes pathologically to a variety of diseases and in minimizing the apoptosis of transplanted myoblasts to treat these and other disease states.

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Abbreviations

FADD:

Fas-associated death domain

TRAIL:

Tumor necrosis factor related apoptosis-inducing ligand

FLIP:

FADD-like IL-1-converting enzyme-inhibitory protein

References

  1. Raff MC (1992) Social controls on cell survival and cell death. Nature 356:397–00

    Article  PubMed  CAS  Google Scholar 

  2. Fidzianska A, Goebel HH (1991) Human ontogenesis 3. Cell death in fetal muscle. Acta Neuropathol (Berl) 81:572–77

    Article  CAS  Google Scholar 

  3. Miller JB, Stockdale FE (1986) Developmental regulation of the multiple myogenic cell lineages of the avian embryo. J Cell Biol 103:2197–208

    Article  PubMed  CAS  Google Scholar 

  4. Sandri MC, Massimino ML, Geromel V, Arslan P (1996) Myoblasts and myotubes in primary cultures deprived of growth factors undergo apoptosis. Basic Appl Myol 6:257–60

    Google Scholar 

  5. Chinni C, de Niese MR, Tew DJ, Jenkins AL, Bottomley SP, Mackie EJ (1999) Thrombin, a survival factor for cultured myoblasts. J Biol Chem 274:9169–174

    Article  PubMed  CAS  Google Scholar 

  6. Wang J, Walsh K (1996) Resistance to apoptosis conferred by Cdk inhibitors during myocyte differentiation. Science 273:359–61

    Article  PubMed  CAS  Google Scholar 

  7. Dee K, Freer M, Mei Y, Weyman CM (2002) Apoptosis coincident with the differentiation of skeletal myoblasts is delayed by caspase 3 inhibition and abrogated by MEK-independent constitutive Ras signaling. Cell Death Differ 9:209–18

    Article  PubMed  CAS  Google Scholar 

  8. Mampuru LJ, Chen SJ, Kalenik JL, Bradley ME, Lee TC (1996) Analysis of events associated with serum deprivation-induced apoptosis in C3H/Sol8 muscle satellite cells. Exp Cell Res 226:372–80

    Article  PubMed  CAS  Google Scholar 

  9. Skuk D, Tremblay JP (2000) Progress in myoblast transplantation: a potential treatment of dystrophies. Microsc Res Tech 48:213–22

    Article  PubMed  CAS  Google Scholar 

  10. Bouchentouf M, Benabdallah BF, Tremblay JP (2004) Myoblast survival enhancement and transplantation success improvement by heat-shock treatment in mdx mice. Transplantation 77:1349–356

    Article  PubMed  CAS  Google Scholar 

  11. Olson EN (1992) Interplay between proliferation and differentiation within the myogenic lineage. Dev Biol 154:261–72

    Article  PubMed  CAS  Google Scholar 

  12. Sandri M, Carraro U (1999) Apoptosis of skeletal muscles during development and disease. Int J Biochem Cell Biol 31:1373–390

    Article  PubMed  CAS  Google Scholar 

  13. Sandri M, El Meslemani AH, Sandri C et al (2001) Caspase 3 expression correlates with skeletal muscle apoptosis in Duchenne and facioscapulo human muscular dystrophy. A potential target for pharmacological treatment? J Neuropathol Exp Neurol 60:302–12

    PubMed  CAS  Google Scholar 

  14. Yang Y, Kaul S, Zhang D, Anantharam V, Kanthasamy AG (2004) Suppression of caspase-3-dependent proteolytic activation of protein kinase C delta by small interfering RNA prevents MPP+-induced dopaminergic degeneration. Mol Cell Neurosci 25:406–21

    Article  PubMed  CAS  Google Scholar 

  15. Nagy Z, Simon L (2004) Neuroprotection in ischemic/hypoxic disorders: from the preclinical to the clinical testing. Adv Exp Med Biol 541:39–4

    PubMed  CAS  Google Scholar 

  16. Przedborski S (2004) Programmed cell death in amyotrophic lateral sclerosis: a mechanism of pathogenic and therapeutic importance. Neurologist 10:1–

    Article  PubMed  Google Scholar 

  17. Sheikh MS, Huang Y (2004) Death receptors as targets of cancer therapeutics. Curr Cancer Drug Targets 4:97–04

    Article  PubMed  CAS  Google Scholar 

  18. Debatin KM, Krammer PH (2004) Death receptors in chemotherapy and cancer. Oncogene 23:2950–966

    Article  PubMed  CAS  Google Scholar 

  19. DeChant AK, Dee K, Weyman CM (2002) Raf-induced effects on the differentiation and apoptosis of skeletal myoblasts are determined by the level of Raf signaling: abrogation of apoptosis by Raf is downstream of caspase 3 activation. Oncogene 21:5268–279

    Article  PubMed  CAS  Google Scholar 

  20. Nakanishi K, Sudo T, Morishima N (2005) Endoplasmic reticulum stress signaling transmitted by ATF6 mediates apoptosis during muscle development. J Cell Biol 169:555–60

    Article  PubMed  CAS  Google Scholar 

  21. Kroemer G (1999) Mitochondrial control of apoptosis: an overview. Biochem Soc Symp 66:1–5

    PubMed  CAS  Google Scholar 

  22. Kuwana T, Newmeyer DD (2003) Bcl-2-family proteins and the role of mitochondria in apoptosis. Curr Opin Cell Biol 15:691–99

    Article  PubMed  CAS  Google Scholar 

  23. Orrenius S (2004) Mitochondrial regulation of apoptotic cell death. Toxicol Lett 149:19–3

    Article  PubMed  CAS  Google Scholar 

  24. Thorburn A (2004) Death receptor-induced cell killing. Cell Signal 16:139–44

    Article  PubMed  CAS  Google Scholar 

  25. Salvesen GS, Dixit VM (1997) Caspases: intracellular signaling by proteolysis. Cell 91:443–46

    Article  PubMed  CAS  Google Scholar 

  26. Budihardjo I, Oliver H, Lutter M, Luo X, Wang X (1999) Biochemical pathways of caspase activation during apoptosis. Annu Rev Cell Dev Biol 15:269–90

    Article  PubMed  CAS  Google Scholar 

  27. Li H, Zhu H, Xu CJ, Yuan J (1998) Cleavage of BID by caspase 8 mediates the mitochondrial damage in the Fas pathway of apoptosis. Cell 94:491–01

    Article  PubMed  CAS  Google Scholar 

  28. Luo X, Budihardjo I, Zou H, Slaughter C, Wang X (1998) Bid, a Bcl2 interacting protein, mediates cytochrome c release from mitochondria in response to activation of cell surface death receptors. Cell 94:481–90

    Article  PubMed  CAS  Google Scholar 

  29. Wang J, Guo K, Wills KN, Walsh K (1997) Rb functions to inhibit apoptosis during myocyte differentiation. Cancer Res 57:351–54

    PubMed  CAS  Google Scholar 

  30. Guo K, Walsh K (1997) Inhibition of myogenesis by multiple cyclin-Cdk complexes. Coordinate regulation of myogenesis and cell cycle activity at the level of E2F. J Biol Chem 272:791–97

    Article  PubMed  CAS  Google Scholar 

  31. Lawlor MA, Rotwein P (2000) Coordinate control of muscle cell survival by distinct insulin-like growth factor activated signaling pathways. J Cell Biol 151:1131–140

    Article  PubMed  CAS  Google Scholar 

  32. Lawlor MA, Rotwein P (2000) Insulin-like growth factor-mediated muscle cell survival: central roles for Akt and cyclin-dependent kinase inhibitor p21. Mol Cell Biol 20:8983–995

    Article  PubMed  CAS  Google Scholar 

  33. Lawlor MA, Feng X, Everding DR, Sieger K, Stewart CE, Rotwein P (2000) Dual control of muscle cell survival by distinct growth factor-regulated signaling pathways. Mol Cell Biol 20:3256–265

    Article  PubMed  CAS  Google Scholar 

  34. Reuveny M, Heller H, Bengal E (2004) RhoA controls myoblast survival by inducing the phosphatidylinositol 3-kinase-Akt signaling pathway. FEBS Lett 569:129–34

    Article  PubMed  CAS  Google Scholar 

  35. Jin Z, El-Deiry WS (2005) Overview of cell death signaling pathways. Cancer Biol Ther 4:139–63

    Article  PubMed  CAS  Google Scholar 

  36. Chinnaiyan AM, O’Rourke K, Tewari M, Dixit VM (1995) FADD, a novel death domain-containing protein, interacts with the death domain of Fas and initiates apoptosis. Cell 81:505–12

    Article  PubMed  CAS  Google Scholar 

  37. Ray S, Almasan A (2003) Apoptosis induction in prostate cancer cells and xenografts by combined treatment with Apo2 ligand/tumor necrosis factor-related apoptosis-inducing ligand and CPT-11. Cancer Res 63:4713–723

    PubMed  CAS  Google Scholar 

  38. Dee K, DeChant A, Weyman CM (2003) Differential signaling through NFkappaB does not ameliorate skeletal myoblast apoptosis during differentiation. FEBS Lett 545:246–52

    Article  PubMed  CAS  Google Scholar 

  39. Kawahara A, Enari M, Talanian RV, Wong WW, Nagata S (1998) Fas-induced DNA fragmentation and proteolysis of nuclear proteins. Genes Cells 3:297–06

    Article  PubMed  CAS  Google Scholar 

  40. Wolfman JC, Palmby T, Der CJ, Wolfman A (2002) Cellular N-Ras promotes cell survival by downregulation of Jun N-terminal protein kinase and p38. Mol Cell Biol 22:1589–606

    Article  PubMed  CAS  Google Scholar 

  41. Almasan A, Ashkenazi A (2003) Apo2L/TRAIL: apoptosis signaling, biology, and potential for cancer therapy. Cytokine Growth Factor Rev 14:337–48

    Article  PubMed  CAS  Google Scholar 

  42. MacFarlane M, Ahmad M, Srinivasula SM, Fernandes-Alnemri T, Cohen GM, Alnemri ES (1997) Identification and molecular cloning of two novel receptors for the cytotoxic ligand TRAIL. J Biol Chem 272:25417–5420

    Article  PubMed  CAS  Google Scholar 

  43. Roth W, Reed JC (2004) FLIP protein and TRAIL-induced apoptosis. Vitam Horm 67:189–06

    Article  PubMed  CAS  Google Scholar 

  44. Zhang K, Kaufman RJ (2006) The unfolded protein response: a stress signaling pathway critical for health and disease. Neurology 66:S102–S109

    Article  PubMed  CAS  Google Scholar 

  45. Wu GS, Burns TF, McDonald ER 3rd et al (1997) KILLER/DR5 is a DNA damage-inducible p53-regulated death receptor gene. Nat Genet 17:141–43

    Article  PubMed  CAS  Google Scholar 

  46. Burns TF, Bernhard EJ, El-Deiry WS (2001) Tissue specific expression of p53 target genes suggests a key role for KILLER/DR5 in p53-dependent apoptosis in vivo. Oncogene 20:4601–612

    Article  PubMed  CAS  Google Scholar 

  47. Cerone MA, Marchetti A, Bossi G, Blandino G, Sacchi A, Soddu S (2000) p53 is involved in the differentiation but not in the differentiation-associated apoptosis of myoblasts. Cell Death Differ 7:506–08

    Article  PubMed  CAS  Google Scholar 

  48. Algeciras-Schimnich A, Griffith TS, Lynch DH, Paya CV (1999) Cell cycle-dependent regulation of FLIP levels and susceptibility to Fas-mediated apoptosis. J Immunol 162:5205–211

    PubMed  CAS  Google Scholar 

  49. Mercer SE, Ewton DZ, Deng X, Lim S, Mazur TR, Friedman E (2005) Mirk/Dyrk1B mediates survival during the differentiation of C2C12 myoblasts. J Biol Chem 280:25788–5801

    Article  PubMed  CAS  Google Scholar 

  50. Peschiaroli A, Figliola R, Coltella L et al (2002) MyoD induces apoptosis in the absence of RB function through a p21(WAF1)-dependent re-localization of cyclin/cdk complexes to the nucleus. Oncogene 21:8114–127

    Article  PubMed  CAS  Google Scholar 

  51. Fukazawa T, Fujiwara T, Uno F et al (2001) Accelerated degradation of cellular FLIP protein through the ubiquitin-proteasome pathway in p53-mediated apoptosis of human cancer cells. Oncogene 20:5225–231

    Article  PubMed  CAS  Google Scholar 

  52. Zhang HG, Wang J, Yang X, Hsu HC, Mountz JD (2004) Regulation of apoptosis proteins in cancer cells by ubiquitin. Oncogene 23:2009–015

    Article  PubMed  CAS  Google Scholar 

  53. Suhara T, Mano T, Oliveira BE, Walsh K (2001) Phosphatidylinositol 3-kinase/Akt signaling controls endothelial cell sensitivity to Fas-mediated apoptosis via regulation of FLICE-inhibitory protein (FLIP). Circ Res 89:13–9

    Article  PubMed  CAS  Google Scholar 

  54. Skurk C, Maatz H, Kim HS et al (2004) The Akt-regulated forkhead transcription factor FOXO3a controls endothelial cell viability through modulation of the caspase-8 inhibitor FLIP. J Biol Chem 279:1513–525

    Article  PubMed  CAS  Google Scholar 

  55. Karasarides M, Dee K, Schulman D, Wolfman A, Weyman CM (2006) Active Ras-induced effects on skeletal myoblast differentiation and apoptosis are independent of constitutive PI3-kinase activity. Cell Biol Int

  56. Panka DJ, Mano T, Suhara T, Walsh K, Mier JW (2001) Phosphatidylinositol 3-kinase/Akt activity regulates c-FLIP expression in tumor cells. J Biol Chem 276:6893–896

    Article  PubMed  CAS  Google Scholar 

  57. Wang Q, Wang X, Hernandez A, Hellmich MR, Gatalica Z, Evers BM (2002) Regulation of TRAIL expression by the phosphatidylinositol 3-kinase/Akt/GSK-3 pathway in human colon cancer cells. J Biol Chem 277:36602–6610

    Article  PubMed  CAS  Google Scholar 

  58. Gilley J, Coffer PJ, Ham J (2003) FOXO transcription factors directly activate bim gene expression and promote apoptosis in sympathetic neurons. J Cell Biol 162:613–22

    Article  PubMed  CAS  Google Scholar 

  59. Cretney E, Takeda K, Yagita H, Glaccum M, Peschon JJ, Smyth MJ (2002) Increased susceptibility to tumor initiation and metastasis in TNF-related apoptosis-inducing ligand-deficient mice. J Immunol 168:1356–361

    PubMed  CAS  Google Scholar 

  60. Finnberg N, Gruber JJ, Fei P et al (2005) DR5 knockout mice are compromised in radiation-induced apoptosis. Mol Cell Biol 25:2000–013

    Article  PubMed  CAS  Google Scholar 

  61. Zhang J, Cado D, Chen A, Kabra NH, Winoto A (1998) Fas-mediated apoptosis and activation-induced T-cell proliferation are defective in mice lacking FADD/Mort1. Nature 392:296–300

    Article  PubMed  CAS  Google Scholar 

  62. Yeh WC, Pompa JL, McCurrach ME et al (1998) FADD: essential for embryo development and signaling from some, but not all, inducers of apoptosis. Science 279:1954–958

    Article  PubMed  CAS  Google Scholar 

  63. Yeh WC, Itie A, Elia AJ et al (2000) Requirement for Casper (c-FLIP) in regulation of death receptor-induced apoptosis and embryonic development. Immunity 12:633–42

    Article  PubMed  CAS  Google Scholar 

  64. Varfolomeev EE, Schuchmann M, Luria V et al (1998) Targeted disruption of the mouse Caspase 8 gene ablates cell death induction by the TNF receptors, Fas/Apo1, and DR3 and is lethal prenatally. Immunity 9:267–76

    Article  PubMed  CAS  Google Scholar 

  65. Ellis RE, Yuan JY, Horvitz HR (1991) Mechanisms and functions of cell death. Annu Rev Cell Biol 7:663–98

    Article  PubMed  CAS  Google Scholar 

  66. Taylor DA, Atkins BZ, Hungspreugs P et al (1998) Regenerating functional myocardium: improved performance after skeletal myoblast transplantation. Nat Med 4:929–33

    Article  PubMed  CAS  Google Scholar 

  67. Scorsin M, Hagege A, Vilquin JT et al (2000) Comparison of the effects of fetal cardiomyocyte and skeletal myoblast transplantation on postinfarction left ventricular function. J Thorac Cardiovasc Surg 119:1169–175

    Article  PubMed  CAS  Google Scholar 

  68. Suzuki K, Murtuza B, Smolenski RT et al (2001) Cell transplantation for the treatment of acute myocardial infarction using vascular endothelial growth factor-expressing skeletal myoblasts. Circulation 104:I207–I212

    Article  PubMed  CAS  Google Scholar 

  69. Skuk D, Tremblay JP (2003) Cell therapies for inherited myopathies. Curr Opin Rheumatol 15:723–29

    Article  PubMed  Google Scholar 

  70. Skuk D, Tremblay JP (2003) Myoblast transplantation: the current status of a potential therapeutic tool for myopathies. J Muscle Res Cell Motil 24:285–00

    Article  PubMed  CAS  Google Scholar 

  71. Skuk D, Caron NJ, Goulet M, Roy B, Tremblay JP (2003) Resetting the problem of cell death following muscle-derived cell transplantation: detection, dynamics and mechanisms. J Neuropathol Exp Neurol 62:951–67

    PubMed  CAS  Google Scholar 

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Acknowledgment

We thank A. Almasan (Learner Research Institute, Cleveland Clinic), V. Dixit (Genetech, Inc.) and E.S.Alnemri (Thomas Jefferson University) for generously providing the dnFADD and dnDR5 constructs. This work was supported by an NIH RO1 CA84212 awarded to C.M. Weyman.

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  1. Department of Biological, Geological, and Environmental Sciences, Cleveland State University, Cleveland, OH, 44115

    J. O’Flaherty, Y. Mei, M. Freer & C. M. Weyman

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  1. J. O’Flaherty
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  2. Y. Mei
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  3. M. Freer
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  4. C. M. Weyman
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Correspondence to C. M. Weyman.

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O’Flaherty, J., Mei, Y., Freer, M. et al. Signaling through the TRAIL receptor DR5/FADD pathway plays a role in the apoptosis associated with skeletal myoblast differentiation. Apoptosis 11, 2103–2113 (2006). https://doi.org/10.1007/s10495-006-0196-4

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