title: Biological effects of a de novo designed myxoma virus peptide analogue: evaluation of cytotoxicity on tumor cells.
abstract: BACKGROUND
  The Resonant Recognition Model (RRM) is a physico-mathematical model that interprets protein sequence linear information using digital signal processing methods. In this study the RRM concept was employed for structure-function analysis of myxoma virus (MV) proteins and the design of a short bioactive therapeutic peptide with MV-like antitumor/cytotoxic activity.
  
  
  RESULTS
  The analogue RRM-MV was designed by RRM as a linear 18 aa 2.3 kDa peptide. The biological activity of this computationally designed peptide analogue against cancer and normal cell lines was investigated. The cellular cytotoxicity effects were confirmed by confocal immunofluorescence microscopy, by measuring the levels of cytoplasmic lactate dehydrogenase (LDH) and by Prestoblue cell viability assay for up to 72 hours in peptide treated and non-treated cell cultures. Our results revealed that RRM-MV induced a significant dose and time-dependent cytotoxic effect on murine and human cancer cell lines. Yet, when normal murine cell lines were similarly treated with RRM-MV, no cytotoxic effects were observed. Furthermore, the non-bioactive RRM designed peptide RRM-C produced negligible cytotoxic effects on these cancer and normal cell lines when used at similar concentrations. The presence/absence of phosphorylated Akt activity in B16F0 mouse melanoma cells was assessed to indicate the possible apoptosis signalling pathway that could be affected by the peptide treatment. So far, Akt activity did not seem to be significantly affected by RRM-MV as is the case for the original viral protein.
  
  
  CONCLUSIONS
  Our findings indicate the successful application of the RRM concept to design a bioactive peptide analogue (RRM-MV) with cytotoxic effects on tumor cells only. This 2.345 kDa peptide analogue to a 49 kDa viral protein may be suitable to be developed as a potential cancer therapeutic. These results also open a new direction to the rational design of therapeutic agents for future cancer treatment.
authors.affiliation: School of Applied Sciences, Science Engineering and Health College, RMIT University, Melbourne, Australia. taghrid.istivan@rmit.edu.au
authors.forename: Taghrid S
authors.initials: TS
authors.lastname: Istivan
authors.forename: Elena
authors.initials: E
authors.lastname: Pirogova
authors.forename: Emily
authors.initials: E
authors.lastname: Gan
authors.forename: Nahlah M
authors.initials: NM
authors.lastname: Almansour
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Coloe
authors.forename: Irena
authors.initials: I
authors.lastname: Cosic
mesh_terms: D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D019295:Computational Biology; D005455:Fluorescent Antibody Technique; D006801:Humans; D051379:Mice; D018613:Microscopy, Confocal; D008958:Models, Molecular; D009233:Myxoma virus / Q000737:chemistry* / Q000187:drug effects; D009369:Neoplasms / Q000473:pathology*; D010455:Peptides / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D011419:Propidium / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry / Q000494:pharmacology
chemical_list: D017304:Annexin A5; D010455:Peptides; D014764:Viral Proteins; D011419:Propidium; D051057:Proto-Oncogene Proteins c-akt