title: Improving the therapeutic potential of endostatin by fusing it with the BAX BH3 death domain.
abstract: Endostatin (ES) inhibits angiogenesis, reducing tumor growth in animal models. However, it has low therapeutic effect in human clinical trials. BAX is a member of the BCL-2 family of proteins; its proapoptotic (BH3) domain interacts with other members of the family in the cytoplasm, to induce apoptosis. Here, we fused the BAX BH3 domain with murine ES, to enhance ES potency. Endothelial cells specifically internalize the fusion protein ES-BAX. The presence of the BAX domain enhances endothelial cell death by apoptosis by 1.8-fold and diminishes microvessel outgrowth in the rat aortic ring assay by 6.5-fold. Daily injections of 15 μg of ES-BAX/g in tumor-bearing mice reduce tumor weight by 86.9% as compared with ES-treated animals. Co-immunoprecipitation assays confirmed that ES-BAX interacts with members of the BCL-2 family. Also, ES interacts with BCL-2, BCL-XL, and BAK in endothelial cell lysates, suggesting a potential new mechanism for the apoptosis induction by ES. The superiority of the ES-BAX antiangiogenic effect indicates that this fusion protein could be a promising therapeutic alternative to treat cancer.
authors.affiliation: Centro de Biotecnologia, Instituto de Pesquisas Energéticas e Nucleares - IPEN - CNEN/SP, São Paulo, Brazil.
authors.forename: R M
authors.initials: RM
authors.lastname: Chura-Chambi
authors.affiliation: Centro de Biotecnologia, Instituto de Pesquisas Energéticas e Nucleares - IPEN - CNEN/SP, São Paulo, Brazil.
authors.forename: M H
authors.initials: MH
authors.lastname: Bellini
authors.affiliation: Laboratório de Investigação Médica LIM62, Faculdade de Medicina da, Universidade de São Paulo, São Paulo, Brazil.
authors.forename: J F
authors.initials: JF
authors.lastname: Jacysyn
authors.affiliation: Departamento de Radiologia e Oncologia, Centro de Investigação Translacional em Oncologia, Instituto do Câncer do Estado de São Paulo, São Paulo, Brazil.
authors.forename: L N
authors.initials: LN
authors.lastname: Andrade
authors.affiliation: Departamento de Imunologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, Brazil.
authors.forename: L P
authors.initials: LP
authors.lastname: Medina
authors.affiliation: Laboratório de Imunopatologia, Instituto Butantan, São Paulo, Brazil.
authors.forename: A R B
authors.initials: AR
authors.lastname: Prieto-da-Silva
authors.affiliation: Departamento de Imunologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, Brazil.
authors.forename: G P
authors.initials: GP
authors.lastname: Amarante-Mendes
authors.affiliation: Centro de Biotecnologia, Instituto de Pesquisas Energéticas e Nucleares - IPEN - CNEN/SP, São Paulo, Brazil.
authors.forename: L
authors.initials: L
authors.lastname: Morganti
mesh_terms: D000595:Amino Acid Sequence; D020533:Angiogenesis Inhibitors / Q000627:therapeutic use / Q000633:toxicity*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D045744:Cell Line, Tumor; D043169:Endostatins / Q000235:genetics / Q000627:therapeutic use / Q000633:toxicity*; D004926:Escherichia coli / Q000378:metabolism; D007680:Kidney Neoplasms / Q000188:drug therapy; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000627:therapeutic use / Q000633:toxicity; D014184:Transplantation, Homologous; D051028:bcl-2-Associated X Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D020533:Angiogenesis Inhibitors; D043169:Endostatins; D011993:Recombinant Fusion Proteins; D051028:bcl-2-Associated X Protein