Review Article

Bacteriophages in Cancer Biology and Therapies

Swapnil Ganesh Sanmukh and Sérgio Luis Felisbino*
Laboratory of Extracellular Matrix Biology, Department of Morphology, Institute of Biosciences Botucatu, Sao Paulo State University, Sao Paulo, Brazil


*Corresponding author: Sérgio L Felisbino, Laboratory of Extracellular Matrix Biology, Department of Morphology, Institute of Biosciences Botucatu, Sao Paulo State University, Sao Paulo, Brazil


Published: 16 May, 2017
Cite this article as: Sanmukh SG, Felisbino SL. Bacteriophages in Cancer Biology and Therapies. Clin Oncol. 2017; 2: 1295.

Abstract

Cancer is one of the most challenging disease because of its unique individual identity and nonuniform progression within the same host. This confronting situation demands individual treatment therapies namely “personalized medicine”. Besides the improved success of available cancer treatments in the majority of cancer cases, for many of them, they have proved ineffective as the cancer cells evolve with treatment and show adaptive behavior. Bacteriophage (phage) therapy and phage-based therapies have gained special attention as a promising approach for cancer therapy beyond their action against multi-drug resistant bacterial infections. Considering the remarkable development in the phage research, mainly in biomedical sciences, molecular biology and nanobiotechnology; until now few conceptual reports about cancer and bacteriophage interactions and/or detailed study for phage-based therapies in cancer are available. These kind of information are valuable for reducing prejudices and to fundament discussions at regulatory agencies and departments for its safe use in humans for cancer therapy. Here we have tried to summarize the important works carried out about phage-cancer research and provide updated information about its ongoing progress and possible future perspectives.
Keywords: Cancer; Bacteriophage; Nanobiotechnology; Regulatory agencies


Abbreviations

KGD Motifs: Tripeptide Lysine-Glycine-Asparagine Motifs; HAP1 Phage: A T4 Sub-Strain With A High Affinity To Melanoma Cells; IL: Interleukin; PMN: Polymorphonuclear Leukocytes; RGD: Tripeptide; Arginine-Glycine-Asparagine; HRP: Horseradish Peroxidase; PC3: Prostate Cancer Cell Line; Grp78: Glucose Regulated Protein; Hsp70: Heat Shock Protein 70; AAVP: Adeno-Associated Virus/Phage; IF: Interferon; USFDA: United States Food And Drug Administration; EMA: European Medicines Agency; IND: Investigational New Drug; CTA: Clinical Trial Application; Tsips: Tumor- Specific Internalizing Peptides; Thps: Tumor Homing Peptides


Introduction

Presently, various types of virus are used for broad range applications in biomedical fields including many mammalian and bacterial viruses (bacteriophages) [1-3]. Among them, bacteriophages have been explored for their range of applications since its discovery in 1915. Besides being a “rosette stone” for molecular biology [4] they have shown various promising application including phage therapy [5-8], phage display (expression of proteins or peptides on the phage capsid) [9-13]| nanoparticles for practical applications in physical and material sciences [10], and industrial applications [14]. The rise of antibiotic resistant bacteria has increased our interest to focus on bacteriophages application as a potential treatment method for bacterial disinfection through phage therapy and phage enzyme applications [15]. There is a need for intensive research on bacteriophages and their role in cancer detection, treatment, diagnosis, etc. as reported recently by researchers worldwide [16]. The phages have been identified as a genetically modifiable molecule in various areas including medicine [17]..
Considering the interest exhibited by various researchers through direct or indirect approaches for bacteriophage applications in medical sciences such as cancer research in last few decades, it is very essential to explore it further to know in which way bacteriophages may prove useful. The possible specificity of bacteriophage for certain cell and tissue components makes them a crucial candidate against cancer. Irrespective to the wide use of bacteriophages for their potential to kill many species of pathogenic bacteria through phage therapies; Nowadays, approaches for using bacteriophages against cancer cells are emerging a lot and further understanding on how mammalian cells responds in vivo to these phage agents with conjugates or through modification requires serious attention [18].


Figure 1

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Figure 1
Ontology of bacteriophage applications for cancer biology and treatment.

History of Bacteriophage in Cancer Research

Since, bacteriophages had proved to be one of the most important tools for various breakthroughs in molecular biology [19,20]. They are mostly studied for their derived applications worldwide along with their lytic life cycle which is well known and their lytic enzymes responsible for bacterial lyses events. The therapeutic potential of phages lies in their structure itself. Its minute and homogeneous size makes it the most capable nanoparticle for drug delivery as well as for various others purposes including phage display and targeting. Their capsid can be used directly to induce immune response naturally (due to their ubiquitous nature) or artificially (through genetic or protein engineering) in any organisms to express desired proteins on its surface to trigger immune response[21]. Due to the limitations of the current review article, we have restricted the broad applications of bacteriophages and just summarized the history of bacteriophage applications in cancer research and their possible outcome in Table 1.


Bacteriophage Interacts with Cancer Cell!

Although some studies have been conducted to know how the bacteriophages can interact with cancer cells through in vivo and in in vitro studies, their impact on influencing host immune response as well as cancer cell physiology are less understood. In 1958, in-vivo and in-vitro demonstration of bacteriophage binding to the cancer cells was reported [22-24]. Similarly, the possible interactions between phage capsid proteins (KGD motifs) as well as β-1 and β-3 integrin receptors on target cancer cells was also confirmed along with the binding mechanism of bacteriophage T4 to Melanoma cells [25,26]. The most puzzling aspect of cancer cell is some of their behaviors, which are similar to a viral infected cell including apoptosis, cell cycle deregulation, metastasis, etc. [27]. Moreover, the 'oncogene', the genes which are attributed to most of the cancers have their origin from viruses. It is by now well established concept that cancer is a systemic biology disease and hence has a very deep root at the gene level [28]. Taking into account the inert properties of phages due to their non pathogenic nature for mammalian host, they can be explored more extensively for further studies for their applications in cancer research.
Current gold standard treatment for cancer includes surgery, chemotherapy, radiation therapy and hormonal therapy. Other advanced treatment methods have shown promising response such as immunotherapy which includes tumor-targeting mAbs, dendritic cell-based interventions, peptide-based vaccines, immunostimulatory cytokines, immuno-modulatory monoclonal antibody, pattern recognition receptor agonists, immunogenic cell death inducers, etc. [29-33], are the emerging nanobiotechnologies recently developed for the detection and treatment of different cancers. Even though these methods are responsible for favorable response in cancer patients, patients now have the option for a personalized medicine approach [34], in which precise knowledge of the tumor molecular signature is accessible making personalized medicine mandatory [35]. In this scenario, bacteriophage, being a nanoparticle, can best fit to be a unique component for various molecular therapies and can be explored as a nanobiomedicine for cancer-treatment.


Why Bacteriophages for Cancer?

Bacteriophages are the most abundant entities in the biosphere that leads to its licensing for human use, mostly due to its non pathogenic nature and inert properties [36,37]. Being ubiquitous, we are frequently consuming large amount of phage populations through different sources [38]. Their detection in saliva, serum and stool samples confirms this finding and validates this observation [39-41]. Various results which have been published earlier showing wide applications of bacteriophages in cancer research and newly proposed applications seems to be very exciting (Figure 1). Some of the most preliminary and established bacteriophage applications are discussed below for getting an in depth idea about its current status and possible future applications.


Bacteriophage as a Natural and Involuntary Medicine

Bacteriophages seem to be an involuntary medicine as they are circulating in the mammalian body controlling invading pathogens and regulating our immune system naturally [42-44]. Furthermore, they can block β3 integrin activity on neoplastic cells thus preventing growth and metastasis formation [42]. Moreover, they also restrict angiogenesis in the developing cancerous tissues or organs and prevent metastasis by inhibiting the adhesion of platelets and T-cells to the fibrinogens [42,45,46]. As initially, it was noticed that phage phagocytes is mostly occurs in liver and spleen [47], hence it can be said that being antigenic in nature they naturally behave as an antigen presenting cells and constantly act as a source for antibody production. Moreover, some phages have been observed to induce the secretion of interleukins (eg. IL-12) and interferons (eg. IF-c) leading to the destruction of tumor tissues through cytokines; which initiate the cascade for neutrophil activation releasing reactive oxidative species (ROS) and other cytotoxic agents, ultimately leading to tissue damage [48]. In our ongoing studies, based on our preliminary results related to natural bacteriophage and Prostate Cancer cell (PC3) interactions it was observed that phages can stimulate internal cellular signaling and were responsible for gene expression alterations. The increased expressions for mRNAs corresponding to integrins (namely integrin αV, α5, β3 and β5) were detected significantly though quantitative PCR along with increase in AKT and PI3K signaling within these cells. Interestingly, we also got down-regulation for Hsp27 and Hsp90 genes after phage treatment, which are on contrary highly expressed in cancer cells and are responsible for migration, proliferation and prevention of apoptosis through survivin up regulation (unpublished). This broad phage binding characteristic as well as its role in specific gene expression mostly through their proteinaceous external coat represents a natural way of phage display.


Table 1

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Table 1
Time line for phage-cancer studies.

Bacteriophage for Gene Delivery, Gene Therapy, Cancer Detection, Targeting and Imaging

Recently, it has been showed that, bacteriophages can be harnessed for their potential in gene delivery to eukaryotic cells through transection of toxic genes to cancer cells. Such gene delivery along with chemotherapeutic agents have great prospective in the near future [49-51]. Similarly, phage display is useful in cancer treatment because it is easy to modify the phage by gene insertion to express desired anticancer proteins and/or peptides on its capsids showing anticancer activities or induce immune response [52].
Similarly, various homing peptides have been reported against cancer specific cells, tissues and organs. The M13/T7 phages were used for tumor cell targeting through tumor homing peptides identification (eg. RGD-4C Peptide) on the cancer cells and tissue specific endothelial cells [53-58]. Moreover, organ specific targeting has been reported to be achieved by using organ homing peptides and antigen detection approaches (eg. prostate-specific membrane antigen) by using M13 as well as other phages [52,59-61]. It was also demonstrated that anticancer drugs efficiency against breast cancer was increased when homing peptides were used with integrins for drug delivery without much toxic effect [62,63]. Identification of tumour homing peptides and organ homing peptides can be helpful for targeting cancer cells, antigen detection and drug delivery using nanoparticles [61,62,64-66]. This approach facilitates the specific targeting of cancer cells, which can be optimized for targeted treatment or drug delivery through bacteriophages.
Nowadays, various databases and online tools are available for predicted, experimentally validated and manually curated comprehensive database for tumor homing and internalizing peptides (Table 2). Many Tumor-Specific Internalizing Peptides (TSIPs) or Tumor Homing Peptides (THPs) have been reported by using in vitro and in vivo phage display technologies; which specifically target tumor along with its microenvironments (eg., tumor vasculature) through their systematic delivery. Such internalizing peptides are of 3 to 15 amino acids length and are smaller than tumor homing peptides. Screening of proteins or peptides for protein-protein interactions can be carried out by displaying them on the surface of a phage through phage coat proteins for identification of internalizing or homing peptides [66].
Furthermore, promiscuous tumor targeting phage proteins have been used for specifically targeting different cancer cells; It is reported that intercellular accumulation of phage clones in cytoplasm, mitochondria as well as nuclear region can be utilized as a potential ligands for intracellular drug delivery and/or molecular imaging techniques as they have few hours of pronuclear accumulation time [67].
Gene therapy is a technique in which functional gene is delivered or introduced into the target cells to restore, over express or inhibit desired gene products [68], which is now used to treat different cancers (e.g., peroxidases whose over expression can lead to the death of cancer cells). Similarly, telomerase is extremely dynamic in most of the cancer and fetal cells, but is almost undetectable in normal tissues hence can be applicable as a good tool for targeting cancer cells [69,70]. Furthermore, glucose regulated protein (Grp78) categorized within the heat shock protein (Hsp70) family can be used as a biomarker in stressed cells such as in tumors as a target for drug delivery or gene therapy by phages [71-73]. The chimeric phage vector, named Adeno-Associated Virus/Phage (AAVP) is a promising candidate in targeted gene therapy [74]. It is a hybrid of eukaryotic adeno associated virus and the filamentous M13 bacteriophage both having single stranded DNA. It expresses 3-5 copies of the cyclic RGD-4C ligand as in RGD-GC peptides on the phage pIII minor coat protein targeting αvβ3-integrin receptor mostly associated with tumor cells [75,76]. Considering this scenario and potential of bacteriophages, they exhibit enormous prospectives in cancer detection, targeting and imaging.


Table 2

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Table 2
Databases and websites for tumor homing/internalizing peptides.

Bacteriophage as an Immunomodulatory

The first study on interactions between bacteriophage and immune response were conducted by Felix d’Herelle demonstrating that phages have the potential to influence immune response [77]. Some phages like phi X174 are considered to be a standard antigen and can trigger humoral responses when administered intravenously; Since then, phage phi X174 have been extensively studied to evaluate humoral immunity in immunosuppressed patients [78,79].
Moreover, due to the omnipresent nature of bacteriophages in the biosphere, phage-neutralizing antibodies are reported to be present in the serum even before phage administration and with systemic administration can increase the phage neutralizing antibody titers [80-83]. Interestingly, considering the antigenic nature of phages, they can stimulate cellular response as whole phage particles and can induce stronger sensitization than their conjugate counterparts as reported in the case of phage sensitized lymphocytes in animals [84]. T-cells do not inactivate phages in vivo, which seems to be a very good indication for further exploring phages for their potential as an immuno modulator [85].
Similarly, rodents show humoral response against phage coat proteins when introduced through oral route [86]. It was also reported that regardless of the route of phage administration, T4 phage particles are cleared considerably from the mice by spleen [86,87]. However, in vivo experimentation studies in mouse have led us to the observation that, T4 phage can be cleared from mouse circulatory system due to removal of Hoc protein (Highly immunogenic outer capsid protein) or defect in their genes [88,89]. As the results seems quite selfcontradictory due to lack of more detailed investigation, there is a need for reevaluation and further in-depth studies for understanding this mechanism. Nevertheless, the therapeutic potential of the phages seems promising and demands urgent attention.
The most interesting thing to be explored is how phages modulate immune system of the diseased host (in vivo) and also cancer cell line responses (in vitro). The first report of phage demonstrating the immune response in animals dated back to 1956, have shown their potential as a new way of treatment methods in the near future [90]. The antibodies produced against phage T4 head proteins (i.e. Hoc and gp23) have been demonstrated to interfere in T4 phage activity [91]. Therefore, it is not a surprise to know that anti-phage antibodies are always present in the healthy individuals [90,92-94]. The studies on bacteriophages and their interaction in mammalian immune system are not yet explored thoroughly. Very few researchers have shown that phages can induce immune response due to their antigenic nature and many more accomplishments are yet to be made [95].


Patents on Bacteriophage Mediated Cancer Treatment

Recent increase in therapeutic approach for cancer treatment have facilitated the use of bacteriophages for cancer detection and treatment as various patents have been granted in the last few decades to phage based cancer technologies. The main aspect of developing the phage based technologies is not just restricted to its nanostructure, immunogenicity or ubiquitous nature but its flexibility, which can be utilized for different applications and therapies. Filamentous phages have been used as a modulator for angiogenesis (Patent: 20150110745) in which phages carrying bacterial lipopollysaccharide endotoxin on its capside surface can be used for treating cancer by modulating angiogenesis process, which can be upregulated or down regulated depending upon the requirement for treating the cancer. Similarly, bacteriophages and their prophage proteins can be used in vitro to express poly-peptides having a proliferation inhibitory activity in the mammalian cell lines as well as it can be used along with pharmaceutical compositions and gene therapeutic approach (Patents: US2009117084, US20050572260 and EP 1853291B1). A patent (US8507445) explaining the methodology as well as targeting peptide composition for diagnosis and therapy of human cancer was also introduced in last decade. Furthermore, patent (WO2002020822 A2) for biopanning and rapid analysis of selective interactive ligands (brasil) was also developed which helps for rapid and efficient separation of specific target binding phages.
Recent patent (Patent: US9034312B2) on extracellular matrix composition for the treatment of cancers looks very promising, as there are various reports of different poly-peptides, mRNAs and microRNAs interfering in cancer metabolic process at cellular and genetic level [52]. Even though few cytotoxic suicide genes are known, there is a requirement for an effective and efficient gene for target oriented killing of cancer cells in a patient (Patent EP 1618886 A1). If we are able to deliver a suicide gene within the specific cancer tissues or organs, nanoparticles like bacteriophage can assist us to specifically detect and target the cancer cells. Another example of combinatory therapy is using miRNAs, which have been reported to modify tumor phenotypes by regulating the genetic mechanism and also regulate cell growth and apoptosis [95]. Considering this potential applicability and possible approaches for targeting cancerous cells, bacteriophages proves to be a good candidate for cancer research encompassing various areas like cancer imaging, detection, targeting and treatment by both in vivo and in vitro applications.


Regulatory Framework for Phage Therapy and Bacteriophage-based Therapy

The bacteriophage therapy is been used for the treatment of various pathogenic bacteria as well as multi drug resistant bacterial infections and they have also shown potential applications for agricultural, animal and human use [96]. However, bacteriophagebased technologies are emerging significantly against various infectious diseases with the advancement in the biotechnological applications and provide options through improved phage-based therapies and diagnostics tools [97].
The main issue is in accepting phage based therapies or products for human use, which are restricted due to regulatory frameworks in different countries. In Poland, the clinical applications of bacteriophages are accepted for treatment as per Polish Law Gazette Number 28 of 1997 and Declaration of Helsinki [98]. Presently, the commercial phage preparations are approved against bacterial pathogens in food for human consumption as per USFDA and are restricted to phage therapy [101,103]. As per standard industrial production criteria, phage-therapy products have been recognized for Investigational New Drug (IND) or Clinical Trial Application (CTA) submission [99-102]. However, bacteriophage based therapies are not yet fully permitted for human use by some important regulatory bodies like United States Food and Drug Administration (USFDA) and the European Medicines Agency (EMA) as very limited randomized controlled clinical trials for bacteriophages have been conducted till date. As an alternative, some US companies have tried to develop phage based products for the decontamination purposes in agricultural sectors [103]. If the possible recommendation of phage based therapies are well accepted, it would be easier to have great awareness for accepting phage based products not only limited for use as an alternative for antibiotics, but also, for other phage derived products for humans including those for cancer treatments.


Future Prospective

The bacteriophages are presently used as a vector for various applications and have been reported to be important constituents of mammalian body playing a vital role in influencing their host immune response. As we know that extracellular matrix composition plays an important role in suppressing cancerous tumor growth, bacteriophages can be used to intensify this effect through their direct or indirect applications [104-109]. The phages can be used to display ploy-peptides and modulate useful constituent of extracellular matrix for anti-cancerous activities by regulating different poly-peptides, mRNAs and microRNAs interfering in cancer mechanisms. Similarly, phage mediated naked eye counting by miRNA molecule in petri dish can be implemented for detection of cancer biomarkers [110-114], which can prove helpful in understanding the cancer cell cycle and pathways. Recently, Anti-bacterial antibody-M13-Dye-probe was used to detect bacterial infections in living host, similar applications can be implemented for detecting cancerous tissues or organs in humans [115]. If we are able to optimize the therapeutic potential of bacteriophages for their direct or indirect applications, we can possibly revolutionize a whole new field of cancer treatment therapy. Bacteriophages can be an efficient cancer research tool and provide a better option for cancer treatment. As observed from the previous research works and results, the applicability of phages in medical field is still not fully explored and hence, in future tremendous breathtaking achievements and technological innovations awaits.


Acknowledgment

This article comprises part of the Ph.D. thesis of SGS, supported by a CAPES funding (processes No. 963-14-2). We would also like to thank Dr. Krystyna Dabrowska from Institute of Immunology and Experimental Therapy at Polish Academy of Science, for providing important literatures for this review.


References

  1. Agrawal P, Bhalla S, Usmani SS, Singh S, Chaudhary K, Raghava GP, et al. CPPsite 2.0: a repository of experimentally validated cell penetrating peptides. Nucleic Acids Research. 2016; 44(D1): D1098-D1103.
  2. Arap W, Kolonin MG, Trepel M, Lahdenranta J, Cardo-Vila M, Giordano RJ, et al. Steps toward mapping the human vasculature by phage display. 2002; 8(2): 121-127.
  3. Arap W, Pasqualini R, Ruoslaht E. Cancer treatment by targeted drug delivery to tumor vasculature in a mouse model. Science. 1998; 279(5349): 377-380.
  4. Arter JA, Diaz JE, Donavan KC, Yuan T, Penner RM, Weiss GA. Virus- polymer hybrid nanowires tailored to detect prostate-specific membrane antigen. Anal Chem. 2012; 84(6): 2776-2783.
  5. Au GG1, Lincz LF, Enno A, Shafren DR. Oncolytic Coxsackievirus A21 as a novel therapy for multiple myeloma. Br J Haematol. 2007; 137(2): 133-141.
  6. Bachrach G, Leizerovici-Zigmond M, Zlotkin A, Naor R, Steinberg D. Bacteriophage isolation from human saliva. Lett Appl Microbiol. 2003; 36(1): 50-53.
  7. Bais S, Bartee E, Rahman MM, McFadden G, Cogle CR. Oncolytic Virotherapy for Hematological Malignancies. Adv Virol. 2012; 2(1): 1-5.
  8. Bardhan N M, Ghosh D, Belcher A M. M13 Virus based detection of Bacterial Infections in Living Hosts. J Biophotonics. 2014; 7(8): 617-623.
  9. Bearden CM, Agarwal A, Book BK, Vieira CA, Sidner RA, Ochs HD, et al. Rituximab inhibits the in vivo primary and secondary antibody responses to a neoantigen, bacteriophage phi X174. Am J Tranplant. 2005; 5(1): 50-57.
  10. Bloch H. Experimental investigation on the relationships between bacteriophages and malignant tumors. Arch Virol. 1940; 1(2): 481-496.
  11. BoratyŠski J, Syper D, Weber-Dabrowska B, Łusiak-Szelachowska M, Poźniak G, Górski A. Preparation of endotoxin-free bacteriophages. Cell Mol Biol Lett. 2004; 9(2): 253-259.
  12. Broccoli D, Young JW, De Lange T. Telomerase activity in normal and malignant hematopoietic cells. Proc Natl Acad Sci USA. 1195; 92(20): 9082-9086.
  13. Burrows FJ and Thorpe PE. Vascular targeting a new approach to the therapy of solid tumors. Pharmac. Ther. 1994; 64: 155-174.
  14. Cao B, Yang M, and Mao C. Phage as a Genetically Modifiable Supramacromolecule in Chemistry, Materials and Medicine. Accounts of Chemical Research. 2016.
  15. Chen PY, Dang X, Klug MT, Qi J, Dorval Courchesne NM, Burpo FJ, et al. Versatile three-dimensional virus-based template for dye-sensitized solar cells with improved electron transport and light harvesting. 2013; 27(8): 6563-6574.
  16. Chen WR, Adams RL, Carubelli R, Nordquist RE. Laser-photosensitizer assisted immunotherapy: a novel modality for cancer treatment. Cancer Lett. 1997; 115(1): 25-30.
  17. Citorik RJ, Mimee M, Lu TK. Bacteriophage-based synthetic biology for the study of infectious diseases. Curr Opin Microbiol. 2014; 19: 59-69.
  18. Clark JR, March JB. Bacterial viruses as human vaccines? Expert Rev Vaccines. 2004; 3(4): 463-476.
  19. Cooper C J, Khan Mirzaei M, Nilsson, A S. Adapting drug approval pathways for bacteriophage-based therapeutics. Front. Microbiol. 2016; 7(7): 1209.
  20. D’Herelle F. Opsonic power of the lysins. In ‘‘Bacteriophage: It’s Role in Immunity’’, p. 125. Williams & Wilkins, Baltimore. 1922.
  21. Dabrowska K, Opolski A, Wietrzyk J, Switala-Jelen K, Boratynski J, Nasulewicz A, et al. Antitumour activity of bacteriophages in murine experimental cancer models caused possibly by inhibition of ß 3 integrin signaling pathway. Acta Virol. 2004; 48(4): 241-248.
  22. Dabrowska K, Opolski A, Wietrzyk J, Switala-Jelen K, Godlewska J, Boratynski J, Syper et al. Anticancer activity of bacteriophage T4 and its mutant HAP1 in mouse experimental tumour models. Anticancer Res. 2004; 24(6): 3991-3995.
  23. Dabrowska K, Switała-Jelen K, Opolski A, Weber-Dabrowska B, Gorski A. Bacteriophage penetration in vertebrates. J Appl Microbiol. 2005; 98(1): 7-13.
  24. Dabrowska Krystyna, Miernikiewicz Paulina, Piotrowicz Agnieszka, Hodyra Katarzyna, Owczarek Barbara, Lecion Dorota, et al. Immunogenicity studies of protein forming the T4 phage head surface. J Virol. 2014; 88(21): 12551-12557.
  25. Denekamp J. Angiogenesis, neovascular proliferation and vascular pathophysiology as targets for cancer therapy. Br J Radiol. 1193; 66(783): 181-196.
  26. DePorter SM. McNaughton BR. Engineered M13 Bacteriophage Nanocarriers for Intracellular Delivery of Exogenous Proteins to Human Prostate Cancer Cells. Bioconjugate Chem. 2014; 25(9): 1620-1625.
  27. Di Giovine M, Salone B, Martina Y, Amati V, Zambruno G, Cundari E. Saggio Binding properties, cell delivery, and gene transfer of adenoviral penton base displaying bacteriophage. Virology. 2001; 282(1): 102-112.
  28. Diamandis M, White NMA, Yousef GM. Personalized Medicine: Marking a New Epoch in Cancer Patient Management. 2010; 8(9): 1175-1187.
  29. Doll TAPF, Raman S, Dey R, Burkhard P. Nanoscale assemblies and their biomedical applications. J R Soc Interface. 2013; 10(80): 657.
  30. Drulis-Kawa Z, Majkowska-Skrobek G, Maciejewska B, Delattre AS, Lavigne, R. Learning from Bacteriophages - Advantages and Limitations of Phage and Phage-Encoded Protein Applications. Current Protein & Peptide Science. 2012; 13(8): 699-722.
  31. Duelli D1, Lazebnik Y. Cell-to-cell fusion as a link between viruses and cancer. Nat Rev Cancer. 2007; 7(12): 968-796.
  32. Edelstein ML, Abedi MR; Wixon J. Gene therapy clinical trials worldwide to 2007- An update. J Gene Med. 2007; 9: 833-842.
  33. Eriksson F, Tsagozis P, Lundberg K, Parsa R, Mangsbo SM, Persson MA, et al. Tumor-specific bacteriophages induce tumor destruction through activation of tumor-associated macrophages. J Immunol. 2009; 182(5): 3105-3111.
  34. Fossati G, Bucknall RC, Edwards SW. Insoluble and soluble immune complexes activate neutrophils by distinct activation mechanisms: changes in functional responses induced by priming with cytokines. Ann Rheum Dis.2002; 61(11): 13-19.
  35. Galluzzi L, Vacchelli E, Bravo-San Pedro JM, Buqué A, Senovilla L, Baracco EE, et al. Classification of current anticancer immunotherapies. Oncotarget. 2014; 5(24): 12472-12508.
  36. Gantzer C, Henny J, Schwartzbrod L. Bacterioides fragilis and Escherichia coli bacteriophages in human faeces. Int J Hyg Environ Health. 2002; 205(4): 324-328.
  37. Garzon R, Marcucci G, Croce CM. Targeting microRNAs in cancer: rationale, strategies and challenges. Nat Rev Drug Discov. 2010; 9(10): 775-789.
  38. Gautam A, Singh H, Tyagi A, Chaudhary K, Kumar R, Kapoor P, Raghava GP. CPPsite: a curated database of cell penetrating peptides. Database (Oxford). 2012.
  39. Geier MR, Trigg ME, Merril CR. Fate of bacteriophage lambda in non-immune germ-free mice. Nature. 1973; 246(5430): 221-223.
  40. Gorski A, Dabrowska K, Switala-Jelen´ K, Nowaczyk M, Weber- Dabrowska B, Boratynski J, et al. New insights into the possible role of bacteriophages in host defense and disease. Med Immunol. 2003; 14: 2.
  41. Gorski A, Kniotek M, Perkowska-Ptasinska A, Mroz A, Przerwa A, Gorczyca W. Bacteriophages and transplantation tolerance. Transplant Proc. 2006; 38(1): 331-333.
  42. Górski A, Wazna E, Dabrowska BW, Dabrowska K, Switała-Jeleń K, Miedzybrodzki R. Bacteriophage translocation. FEMS Immunol Med Microbiol. 2006; 46(3): 313-319.
  43. Gorski A, Weber-Dabrowska B. The potential role of endogenous bacteriophages in controlling invading pathogens. Cell Mol Life Sci. 2005; 62: 511-519.
  44. Goubran HA, Elemary M, Radosevich M, Seghatchian J, El-Ekiaby M, Burnouf T. Impact of Transfusion on Cancer Growth and Outcome. Cancer Growth and Metastasis. 2016.
  45. Gross AL, Gillespie JW, Petrenko VA. Promiscuous tumor targeting phage proteins. Protein Eng Des Sel. 2016; 29(3): 93-103
  46. Hajitou A, Rangel R, Trepel M, Soghomonyan S, Gelovani JG, Alauddin MM, et al. Design and construction of targeted aavp vectors for mammalian cell transduction. Nat Protoc. 2007; 2(3): 523-531.
  47. Hajitou A, Trepel M, Lilley CE, Soghomonyan S, Alauddin MM. Marini FC3rd, Rest el al. A hybrid vector for ligand-directed tumor targeting and molecular imaging. Cell. 2006; 125: 385-398.
  48. Hedstrom SA, Kamme C. Antibodies against staphylococcal bacteriophages in human sera. II. Assay of antibodies in exacerbation and regression of chronic staphylococcal osteomyelitis. Acta Path Microbiol Scand. 1973; 81(6): 749-752.
  49. Henry M, Debarbieux L. Tools from viruses: Bacteriophage successes and beyond. Virology. 2012; 434(2): 151-161.
  50. Hornberg JJ, Bruggeman FJ, Westerhoff HV, Lankelma J. Cancer: a Systems Biology disease. Biosystems. 2006; 83(2-3): 81-90.
  51. http://www.bioprocessintl.com/manufacturing/nonantibody-therapeutics/bacteriophages-an-alternative-to-antibiotics-challenges-and-possible-solutions-for-bringing-them-to-market.
  52. Ibrahim SA, Hassan H, Götte M. MicroRNA-dependent targeting of the extracellular matrix as a mechanism of regulating cell behavior. Biochim Biophys Acta. 2014; 1840(2): 2609-2620.
  53. Inchley CJ. The activity of mouse kupffer cells following intravenous injection of T4 bacteriophage. Clin Exp Immunol. 1969; 5(2):173-187.
  54. Jerne NK, Avegno P. The development of the phage-inactivating properties of serum during the course of specific immunization of an animal: reversible and irreversible inactivation. J Immunol. 1956; 76(3): 200-208.
  55. Jerne NK. The presence in normal serum of specific antibody against bacteriophage T4 and its increase during the earliest stages of immunization. J Immunol. 1956; 76(3): 209-216.
  56. Jin SE, Jin HE, Hong SS. Targeted delivery system of nanobiomaterials in anticancer therapy: from cells to clinics. Biomed Res Int. 2014; 814208(6): 1-23.
  57. Kamme C. Antibodies against staphylococcal bacteriophages in human sera. I. Assay of antibodies in healthy individuals and in patients with staphylococcal infections. Acta Pathol Microbiol Scand (B) Microbiol Immunol. 1973; 81(6): 741-748.
  58. Kantoch M, Mordarski M. Binding of bacterial viruses by cancer cells in vitro. Postepy Hig Med Dosw. 1958; 12(2): 191-192.
  59. Kapoor P, Singh H, Gautam A, Chaudhary K, Kumar R, Raghava GP. TumorHoPe: a database of tumor homing peptides. PLoS One. 2012; 7(4): e35187.
  60. Keen EC. A century of phage research: bacteriophages and the shaping of modern biology. Bioessays. 2015; 37(1): 6-9.
  61. Kim M, Madlambayan GJ, Rahman MM. Myxoma virus targets primary human leukemic stem and progenitor cells while sparing normal hematopoietic stem and progenitor cells. Leukemia. 2009; 23(3): 2313-2317.
  62. Kleinschmidt WJ, Douthart RJ, Murphy EB. Interferon production by T4 coliphage. Nature. 1970; 228(5266): 27-30.
  63. Kniotek M, Ahmed AMA, Dabrowska K, Switala-Jelen K, Opolski A, Gorski A. Bacteriophage interactions with T cells and platelets. In cytokine network, Regulatory, cells signalling, and apoptosis (immunology). Monduzzi Editors, Bologna. 2004; 89-192.
  64. Kucharzewicz-Krukowska A, Slopek S. Immunogenic effects of bacteriophage in patients subjected to phage therapy. Arch Immun Ther Exp. 1987; 35(7): 553-561.
  65. Kurzepa A, Dabrowska K, Skaradziński G, Górski A. Bacteriophage interactions with phagocytes and their potential significance in experimental therapy. Clin Exp Med. 2009; 9(2): 93-100.
  66. Kurzepa-Skaradzinska A, Lusiak-Szelachowska M, Skaradzinski G, Jonczyk-Matysiak E, Weber-Dabrowska B, Zaczek M, et al. Influence of bacteriophage preparations on intracellular killing of bacteria by human phagocytes in vitro. Viral Immunol. 2013; 26(2): 150-162.
  67. Larocca D, Burg MA, Jensen-Pergakes K, Ravey EP, Gonzalez AM, Baird A. Evolving phage vectors for cell targeted gene delivery. Curr Pharm Biotechnol. 2002; 3(1): 45-57.
  68. López MB, Teijeiro A, Rivas J. Magnetic nanoparticle-based hyperthermia for cancer treatment. Reports of Practical Oncology & Radiotherapy 2013; 18(6): 397-400.
  69. Mactier CE, Islam MS2. Haematopoietic stem cell transplantation as first-line treatment in myeloma: a global perspective of current concepts and future possibilities. Oncol Rev. 2012; 6(2): e14.
  70. Matsumoto M, Saeki S, Kajiwara K, Ishihara H, Amano T, Nishizaki T, et al. ERvelopment of brain tumor demarcation technique using two-color laser-induced fluorescence. 2014; 33(4): 260-278.
  71. Merabishvili M, Pirnay JP, Verbeken G, Chanishvili N, Tediashvili M, Lashkhi N, et al. Quality-controlled small-scale production of a well-defined bacteriophage cocktail for use in human clinical trials. PLoS One. 2009; 4(3): e4944.
  72. Merril CR, Friedman TB, Attallah AF, Geier MR, Krell K, Yarkin R. Isolation of bacteriophages from commercial sera. In Vitro. 1972; 8(2): 91-93.
  73. Mohan K, Weiss G A. Engineering chemically modified viruses for prostate cancer cell recognition. Mol. Biosyst. 2015; 11: 3264-3272.
  74. Mohan K, Weiss GA1. Chemically Modifying Viruses for Diverse Applications. ACS Chem Biol. 2016; 11(5): 1167-1179.
  75. Nilsson AS. Phage therapy--constraints and possibilities. Ups J Med Sci. 2014; 119(2): 192-198.
  76. Oliveira H, Sillankorva S, Merabishvili M, Kluskens LD, Azeredo J. Unexploited opportunities for phage therapy. Front Pharmacol. 2015; 6(2): 180.
  77. Pajtasz-Piasecka E, Rossowska J, Dum D, Weber-Dabrowska B, Zablocka A, Gorski A. Bacteriophages support anti-tumor response initiated by DC-based vaccine against murine transplantable colon carcinoma. Immunol Lett. 2008; 116: 24-32.
  78. Pasqualini R, Ruoslahti E. Tissue targeting with phage peptide libraries. Mol Psychiatry. 1996; 1: 423-423.
  79. Pasqualini R. Vascular targeting with phage peptide libraries. Q J Nucl Med. 1999; 43(2): 159-162.
  80. Pelfrene E, Willebrand E2, Cavaleiro Sanches A3, Sebris Z4, Cavaleri M2. Bacteriophage therapy: a regulatory perspective. J Antimicrob Chemother. 2016; 71(8): 2071-2074.
  81. Pirnay JP, Blasdel BG, Bretaudeau L, Buckling A, Chanishvili N, Clark JR, et al. Quality and Safety Requirements for Sustainable Phage Therapy. Products. Pharm. Res. 2015(7): 2173-2179.
  82. Pranjol Md, Zahidul I and Hajitou A. Bacteriophage-Derived Vectors for Targeted Cancer Gene Therapy. Viruses. 2015; (1): 268-284.
  83. Przerwa A, Kniotek M, Nowaczyk M, Weber-Dabrowska B, Switala-Jelen K, Dabrowska K,et al. Bacteriophages inhibit interleukin-2 production by human T lymphocytes. In ‘‘12th Congress of the European Society for Organ Transplantation, Geneva Switzerland. 2005.
  84. Przerwa A, Zimecki M, Switala-Jelen´ K, Dabrowska K, Krawczyk E, Luczak M. Effects of bacteriophages on free radical production and phagocytic functions. Med Microbiol Immunol. 2006; 195(6): 143-150.
  85. Rama AR, Prados J, Melguizo C, Alvarez PJ, Ortiz R, Madeddu R, et al. The phage gene transfection associated to chemotherapeutic agents increases apoptosis in lung and colon cancer cells. Bioeng Bugs. 2011; 2(2): 163-167.
  86. Rivinoja A, Laakkonen P. Identification of homing peptides using the in vivo phage display technology. Methods Mol Biol. 2011; 683(3): 401-415.
  87. Sathaliyawala T, Islam MZ, Li Q, Fokine A, Rossmann MG, Rao VB. Functional analysis of the highly antigenic outer capsid protein, Hoc, a virus decoration protein from T4-like bacteriophages. Mol Microbiol. 2010; 77(2): 444-455.
  88. Schilsky RL. Personalized medicine in oncology: the future is now. Nature Reviews Drug Discovery. 2010; 9(5): 363-366.
  89. Sharma A, Kapoor P, Gautam A, Chaudhary K, Kumar R, Chauhan JS, et al. Computational approach for designing tumor homing peptides. Sci Rep. 2013; 3: 1607.
  90. Shu D, Li H, Shu Y, Xiong G, Carson W E, Haque F, et al. Systemic Delivery of Utilizing RNA Nanotechnology Anti-miRNA for Suppression of Triple Negative Breast Cancer. ACS Nano; Ahead of printing. 2015.
  91. Sinkovics JG, Horvath JC. Natural and genetically engineered viral agents for oncolysis and gene therapy of human cancers. Arch Immunol Ther Exp (Warsz). 2008; 56(1): 3s-59s.
  92. Srivastava AS, Kaido T, Carrier E. Immunological factors that affect the in vivo fate of T7 phage in the mouse. J Virol Methods. 2004; 115(1): 99-104.
  93. Stone R. Stalin's forgotten cure. Science. 2002; 298(5594): 728-731.
  94. Sugahara KN, Teesalu T, Karmali PP, Kotamraju VR, Agemy L, Girard OM, et al. Tissue-penetrating delivery of compounds and nanoparticles into tumors. Cancer Cell. 2009; 16(6): 510-520.
  95. Sulakvelidze A, Alavidze Z, Morris JG Jr. Bacteriophage therapy. Antimicrob Agents Chemother. 2001; 45(3): 649-659.
  96. Srivastava AS, Kaido T, Carrier E. Badanie mechanizmu dziaiania przeciwnowotworowego preparatu fagowego BP T4 u myszy obarczonych przeszczepialnym czerniakiem B16 (In Polish). Dissertation, Polish Academy of Science, Institute of Immunology and Experimental Therapy, Wroclaw, Poland. 2004.
  97. Thiel K. Old dogma, new tricks--21st Century phage therapy. Nat Biotechnol. 2004; 22(1): 31-36.
  98. Thirukkumaran CM, Luider JM, Stewart DA, et al. Reovirus oncolysis as a novel purging strategy for autologous stem cell transplantation. Blood. 2003; 102(1): 377-387.
  99. Torring T, Helmig S, Ogilby PR, Gothelf KV. Singlet Oxygen in DNA Nanotechnology. Acc Chem Res. 2014; 47(2): 1799-1806.
  100. Twort F. An investigation on the nature of ultra-microscopic viruses. Lancet. 1915; 186(4): 4814.
  101. Verbeken G, De Vos D, Vaneechoutte M, Merabishvili M, Zizi M, Pirnay J P. European regulatory conundrum of phage therapy. Future Microbiology. 2007; 2(5): 485-491.
  102. Verbeken G, Pirnay J P, De Vos D, Jennes S, Zizi M, Lavigne R, et al. Call for a dedicated European legal framework for bacteriophage therapy. Arch. Immunol. Ther. Exp. 2014; 62(2): 117-129.
  103. Verbeken G, Pirnay JP, De Vos D, Jennes S, Zizi M, Lavigne R, et al. Optimizing the European Regulatory Framework for Sustainable Bacteriophage Therapy in Human Medicine. Arch Immunol Ther Exp. 2012; 60: 161.
  104. Wang G, Li X, Wang Z. APD2: The updated antimicrobial peptide database and its application in peptide design. Nucleic Acids Research. 2009.
  105. Wang, G Li, X Wang Z. APD3: The antimicrobial peptide database as a tool for research and education. Nucleic Acids Research. 2016; 44.
  106. Wang Z, Wang G. APD: the Antimicrobial Peptide Database. Nucleic Acids Res. 2004; 32: D590-592.
  107. Warner C, Barker N, Lee SW, Perkins E. M13 bacteriophage production for large-scale applications. Bioprocess Biosyst Eng. 2014; 37(10): 2067-2072.
  108. Weber-Dabrowska B, Mulczyk M, Górski A. Bacteriophage therapy of bacterial infections: an update of our institute's experience. Arch Immunol Ther Exp. 2000; 48(6): 547-551.
  109. Wedgwood R J, Ochs H D, Davis SD. The recognition and classification of immunodeficiency diseases with bacteriophage phi X174. Birth Defects Orig Artic Ser. 1975; 11(2): 331-338.
  110. Wenger SL, Turner JH, Petricciani JC. The cytogenetic, proliferative and viability effects of four bacteriophages on human lymphocytes. In Vitro. 1979; 14(6): 543-549.
  111. Yang XL, Fan CH, Zhu HS. Photo-induced cytotoxicity of malonic acid [C60] fullerene derivatives and its mechanism. Toxicology in Vitro. 2002; 16(1): 41-46.
  112. Zagar TM, Oleson JR, Vujaskovic Z, Dewhirst MW, Craciunescu OI, Blackwell KL, et al. Hyperthermia for locally advanced breast cancer. Int J Hyperthermia. 2010; 26(7): 618-624.
  113. Zhang LH, Zhang X. Roles of GRP78 in physiology and cancer. J Cell Biochem. 2010; 110(6): 1299-1305.
  114. Zheng Y, Yin G, Le V, Zhang A, Chen S, Liang X, Liu J. Photodynamic-therapy Activates Immune Response by disrupting Immunity Homeostasis of Tumor Cells, which Generates Vaccine for Cancer Therapy. Int J Biol Sci. 2016; 12(1): 120-132.
  115. Zhou X, Cao P, Zhu Y, Lu W, Gu N and Mao C. Phage-mediated counting by the naked eye of miRNA molecules at attomolar concentrations in a Petri dish. Nature Materials. 2015; 14(10): 1058-1064.