Designed tumor necrosis factor-related apoptosis-inducing ligand variants initiating apoptosis exclusively via the DR5 receptor.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a potential anticancer drug that selectively induces apoptosis in a variety of cancer cells by interacting with death receptors DR4 and DR5. TRAIL can also bind to decoy receptors (DcR1, DcR2, and osteoprotegerin receptor) that cannot induce apoptosis. The occurrence of DR5-responsive tumor cells indicates that a DR5 receptor-specific TRAIL variant will permit tumor-selective therapies. By using the automatic design algorithm FOLD-X, we successfully generated DR5-selective TRAIL variants. These variants do not induce apoptosis in DR4-responsive cell lines but show a large increase in biological activity in DR5-responsive cancer cell lines. Even wild-type TRAIL-insensitive ovarian cancer cell lines could be brought into apoptosis. In addition, our results demonstrate that there is no requirement for antibody-mediated cross-linking or membrane-bound TRAIL to induce apoptosis through DR5.

[1]  D. Baker,et al.  Computational redesign of protein-protein interaction specificity , 2004, Nature Structural &Molecular Biology.

[2]  J. Tschopp,et al.  The molecular architecture of the TNF superfamily. , 2002, Trends in biochemical sciences.

[3]  S. S. Strom,et al.  Differential hepatocyte toxicity of recombinant Apo2L/TRAIL versions , 2001, Nature Medicine.

[4]  D. Lawrence,et al.  Safety and antitumor activity of recombinant soluble Apo2 ligand. , 1999, The Journal of clinical investigation.

[5]  Z. Wang,et al.  Tumoricidal activity of a novel anti-human DR5 monoclonal antibody without hepatocyte cytotoxicity , 2001, Nature Medicine.

[6]  Eugene A Zhukovsky,et al.  Inactivation of TNF Signaling by Rationally Designed Dominant-Negative TNF Variants , 2003, Science.

[7]  L. Serrano,et al.  Prediction of water and metal binding sites and their affinities by using the Fold-X force field. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[8]  J. Tschopp,et al.  The Tumor Necrosis Factor-related Apoptosis-inducing Ligand Receptors TRAIL-R1 and TRAIL-R2 Have Distinct Cross-linking Requirements for Initiation of Apoptosis and Are Non-redundant in JNK Activation* , 2000, The Journal of Biological Chemistry.

[9]  M. Ultsch,et al.  Triggering cell death: the crystal structure of Apo2L/TRAIL in a complex with death receptor 5. , 1999, Molecular cell.

[10]  Karen Billeci,et al.  Receptor-selective Mutants of Apoptosis-inducing Ligand 2/Tumor Necrosis Factor-related Apoptosis-inducing Ligand Reveal a Greater Contribution of Death Receptor (DR) 5 than DR4 to Apoptosis Signaling* , 2005, Journal of Biological Chemistry.

[11]  J. Blenis,et al.  FADD/MORT1 and caspase-8 are recruited to TRAIL receptors 1 and 2 and are essential for apoptosis mediated by TRAIL receptor 2. , 2000, Immunity.

[12]  A. Ashkenazi,et al.  Apo2L/TRAIL and its death and decoy receptors , 2003, Cell Death and Differentiation.

[13]  D. Stuart,et al.  Structure of the TRAIL–DR5 complex reveals mechanisms conferring specificity in apoptotic initiation , 1999, Nature Structural Biology.

[14]  R. Locksley,et al.  The TNF and TNF Receptor Superfamilies Integrating Mammalian Biology , 2001, Cell.

[15]  V. Rybin,et al.  Computer-aided design of a PDZ domain to recognize new target sequences , 2002, Nature Structural Biology.

[16]  Julia M. Shifman,et al.  Modulating calmodulin binding specificity through computational protein design. , 2002, Journal of molecular biology.

[17]  L. Serrano,et al.  Predicting changes in the stability of proteins and protein complexes: a study of more than 1000 mutations. , 2002, Journal of molecular biology.

[18]  D. Lawrence,et al.  Apo2L/TRAIL-dependent recruitment of endogenous FADD and caspase-8 to death receptors 4 and 5. , 2000, Immunity.

[19]  M. Ultsch,et al.  A unique zinc-binding site revealed by a high-resolution X-ray structure of homotrimeric Apo2L/TRAIL. , 2000, Biochemistry.

[20]  G. Screaton,et al.  Following a TRAIL: Update on a ligand and its five receptors , 2004, Cell Research.

[21]  T L Chenevert,et al.  Combined effect of tumor necrosis factor-related apoptosis-inducing ligand and ionizing radiation in breast cancer therapy. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[22]  A. Thorburn,et al.  The C-terminal Tails of Tumor Necrosis Factor-related Apoptosis-inducing Ligand (TRAIL) and Fas Receptors Have Opposing Functions in Fas-associated Death Domain (FADD) Recruitment and Can Regulate Agonist-specific Mechanisms of Receptor Activation* , 2004, Journal of Biological Chemistry.

[23]  M. S. Lee,et al.  Crystal Structure of TRAIL-DR5 Complex Identifies a Critical Role of the Unique Frame Insertion in Conferring Recognition Specificity* , 2000, The Journal of Biological Chemistry.

[24]  J. Tschopp,et al.  TRAIL receptor-2 signals apoptosis through FADD and caspase-8 , 2000, Nature Cell Biology.

[25]  K. Bhalla,et al.  Antileukemic drugs increase death receptor 5 levels and enhance Apo-2L-induced apoptosis of human acute leukemia cells. , 2000, Blood.

[26]  Christina Kiel,et al.  A detailed thermodynamic analysis of ras/effector complex interfaces. , 2004, Journal of molecular biology.

[27]  P. Harbury,et al.  Automated design of specificity in molecular recognition , 2003, Nature Structural Biology.

[28]  Joost Schymkowitz,et al.  Recognizing and defining true Ras binding domains II: in silico prediction based on homology modelling and energy calculations. , 2005, Journal of molecular biology.

[29]  H. Koeppen,et al.  Isotype-Dependent Inhibition of Tumor Growth In Vivo by Monoclonal Antibodies to Death Receptor 4 , 2001, The Journal of Immunology.

[30]  G Vriend,et al.  WHAT IF: a molecular modeling and drug design program. , 1990, Journal of molecular graphics.

[31]  L. Serrano,et al.  Stabilization of TRAIL, an all-beta-sheet multimeric protein, using computational redesign. , 2004, Protein engineering, design & selection : PEDS.

[32]  P. Scheurich,et al.  Differential activation of TRAIL-R1 and -2 by soluble and membrane TRAIL allows selective surface antigen-directed activation of TRAIL-R2 by a soluble TRAIL derivative , 2001, Oncogene.

[33]  C. Smith,et al.  Functional analysis of TRAIL receptors using monoclonal antibodies. , 1999, Journal of immunology.