Identification of Novel Targeting Peptides for PC3 Cells by the Screening of a Phage Display Peptide Library

Authors
1 Department of Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran
2 Department of Medical Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
Abstract
Objective: Prostate cancer is the second cause of cancer-associated death in men. In recent years, targeted therapy for cancer has attracted the attention of researchers. Targeted therapy leads to a decrease in drug adverse effects. Studies indicate that targeting peptides for cancer cells represent valuable tools for diagnostics and therapeutics. Recently, phage display peptide libraries have been used to identify target peptides to a variety of cancer cells. In the current study, we aim to isolate peptides that target PC3 cells (human prostate adenocarcinoma cells).
Methods: Four rounds of subtractive panning on control cells that included 5637 (bladder), Huh-7 (liver), SW480 (colon), AGS (stomach) and human fibroblast normal in addition to four rounds of positive panning on PC3 (target cell) were performed. Polyclonal phage ELISA was used to evaluate the process of enrichment during biopanning. Subsequently, phage clones were randomly selected from titer plates, amplified by plaque-PCR, and their genomic DNA was sequenced. We conducted bioinformatic analysis for further characterization of the isolated peptides.
Results: Several rounds of panning resulted in the enrichment of a number of peptides. The results of polyclonal phage ELISA indicated that the biopanning process was successful. In silico analysis showed the presence of several consensus amino acid motifs in the peptides.
Conclusion: The peptides identified through biopanning can be considered as potential specific binders to PC3 cells. Peptides with specificity binding to target cells can be used for targeted gene and drug delivery to malignant tumor cells. Further analyses of these peptides are required to show their capacity for targeted delivery of various genes and drugs into prostate cancer cells.

Keywords


[1]     Carter HB, Partin AW. Diagnosis and staging of prostate cancer. Campbell’s Urology. 8th ed. Sydney: Elsevier Science Publishers, 2002.
[2]     Jemal A, Murray T, Ward E, Samuels A, Tiwari RC, Ghafoor A, Feuer EJ, Thun MJ. Cancer statistics, 2005. CA Cancer J Clin 2005; 55(1): 10-30.
[3]     Abate-Shen C, Shen MM. Molecular genetics of prostate cancer. Genes Dev 2000; 14(19): 2410-34.
[4]     Mohagheghi MA, Mosavi-Jarrahi A, Malekzadeh R, Parkin M. Cancer incidence in Tehran metropolis: the first report from the Tehran Population-based Cancer Registry, 1998-2001. Arch Iran Med 2009; 12(1): 15-23.
[5]     Frydenberg M, Wijesinha S. Diagnosing prostate cancer - what GPs need to know. Aust Fam Physician 2007; 36(5): 345-7.
[6]     Herschman JD, Smith DS, Catalona WJ. Effect of ejaculation on serum total and free prostate-specific antigen concentrations. Urology 1997; 50(2): 239-43.
[7]     Nadler RB, Humphrey PA, Smith DS, Catalona WJ, Ratliff TL. Effect of inflammation and benign prostatic hyperplasia on elevated serum prostate specific antigen levels. J Urol 1995; 154(2 Pt 1): 407-13.
[8]     Carter HB. Assessing risk: does this patient have prostate cancer? J Natl Cancer Inst 2006; 98(8): 506-7.
[9]     Thompson IM, Pauler DK, Goodman PJ, Tangen CM, Lucia MS, Parnes HL, Minasian LM, Ford LG, Lippman SM, Crawford ED, Crowley JJ, Coltman CA Jr. Prevalence of prostate cancer among men with a prostate-specific antigen level < or =4.0 ng per milliliter. N Engl J Med 2004; 350(22): 2239-46.
[10] Tinzl M, Marberger M, Horvath S, Chypre C. DD3PCA3 RNA analysis in urine--a new perspective for detecting prostate cancer. Eur Urol 2004; 46(2): 182-6.
[11] Fradet Y, Saad F, Aprikian A, Dessureault J, Elhilali M, Trudel C, Mâsse B, Piché L, Chypre C. uPM3, a new molecular urine test for the detection of prostate cancer. Urology 2004; 64(2): 311-5.
[12] Landon LA, Zou J, Deutscher SL. Is phage display technology on target for developing peptide-based cancer drugs? Curr Drug Discov Technol 2004; 1(2): 113-32.
[13] Romanov VI, Durand DB, Petrenko VA. Phage display selection of peptides that affect prostate carcinoma cells attachment and invasion. Prostate 2001; 47(4): 239-51.
[14] Petrenko VA, Jayanna PK. Phage protein-targeted cancer nanomedicines. FEBS Lett 2014; 588(2): 341-9.
[15] Jain RK. The next frontier of molecular medicine: delivery of therapeutics. Nat Med. 1998; 4(6): 655-7.
[16] Poul MA, Marks JD. Targeted gene delivery to mammalian cells by filamentous bacteriophage. J Mol Biol 1999; 288(2): 203-11.
[17] Cwirla SE1, Peters EA, Barrett RW, Dower WJ. Peptides on phage: a vast library of peptides for identifying ligands. Proc Natl Acad Sci U S A 1990; 87(16): 6378-82.
[18] Smith GP. Filamentous fusion phage: novel expression vectors that display cloned antigens on the virion surface. Science 1985; 228(4705): 1315-7.
[19] Brown KC. Peptidic tumor targeting agents: the road from phage display peptide selections to clinical applications. Curr Pharm Des 2010; 16(9): 1040-54.
[20] Ph.D.™-7 Phage Display Peptide Library Instruction Manual at www.neb.com/products/ e8102-phd-7-phage-display-peptide-library. New England Biolabs.
[21] Newton JR, Deutscher SL, In vivo bacteriophage display for the discovery of novel peptide-based tumor-targeting agents. Methods Mol Biol 2009; 504: 275-90.
[22] Newton JR1, Kelly KA, Mahmood U, Weissleder R, Deutscher SL. In vivo selection of phage for the optical imaging of PC-3 human prostate carcinoma in mice. Neoplasia 2006; 8(9): 772-80.
[23] Arap W, Kolonin MG, Trepel M, Lahdenranta J, Cardó-Vila M, Giordano RJ, Mintz PJ, Ardelt PU, Yao VJ, Vidal CI, Chen L, Flamm A, Valtanen H, Weavind LM, Hicks ME, Pollock RE, Botz GH, Bucana CD, Koivunen E, Cahill D, Troncoso P, Baggerly KA, Pentz RD, Do KA, Logothetis CJ, Pasqualini R. Steps toward mapping the human vasculature by phage display. Nat Med 2002; 8(2): 121-7.
[24] Agris PF, Marchbank MT, Newman W, Guenther R, Ingram P, Swallow J, Mucha P, Szyk A, Rekowski P, Peletskaya E, Deutscher SL. Experimental models of protein-RNA interaction: isolation and analyses of tRNA(Phe) and U1 snRNA-binding peptides from bacteriophage display libraries. J Protein Chem 1999; 18(4): 425-35.
[26] Isaacs WB, Carter BS, Ewing CM. Wild-type p53 suppresses growth of human prostate cancer cells containing mutant p53 alleles. Cancer Res 1991; 51(17): 4716-20.
[27] Kevin JH, Richard GV, Hardev SP. Viral Therapy of Cancer. John Wiley & Sons, Ltd. 2008; p: 114-8.
[28] Truong LD, Kadmon D, McCune BK, Flanders KC, Scardino PT, Thompson TC. Association of transforming growth factor-beta with prostate cancer: An immunochemical study. Hum Pathol 1993; 24(1): 4-9.
[29] Kelly KA, Jones DA. Isolation of a colon tumor specific binding peptide using phage display selection. Neoplasia 2003; 5(5): 437-44.
[30] Lee TY, Lin CT, Kuo SY, Chang DK, Wu HC. Peptide-mediated targeting to tumor blood vessels of lung cancer for drug delivery. Cancer Res 2007; 67(22): 10958-65.
[31] Zhang J, Spring H, Schwab M. Neuroblastoma tumor cell-binding peptides identified through random peptide phage display. Cancer Lett 2001; 171(2): 153-64.
[32] Lee SM, Lee EJ, Hong HY, Kwon MK, Kwon TH, Choi JY, Park RW, Kwon TG, Yoo ES, Yoon GS, Kim IS, Ruoslahti E, Lee BH. Targeting bladder tumor cells in vivo and in the urine with a peptide identified by phage display. Mol Cancer Res 2007; 5(1): 11-9.
[33] Zhang Y, Chen J, Zhang Y, Hu Z, Hu D, Pan Y, Ou S, Liu G, Yin X, Zhao J, Ren L, Wang J. Panning and identification of a colon tumor binding peptide from a phage display peptide library. J Biomol Screen 2007; 12(3): 429-35.
[34] Bakhshinejad B1, Karimi M, Sadeghizadeh M. Bacteriophages and medical oncology: targeted gene therapy of cancer. Med Oncol 2014; 31(8): 110.
[35] Pande J, Szewczyk MM, Grover AK. Phage display: concept, innovations, applications and future. Biotechnol Adv 2010; 28(6): 849-58.