Title: Use of oncolytic viruses as therapeutic strategy for cancer A dissertation presented by: Name: Farhath Jabien Student

number: S87 !"#7J U$%: & '(!"7 %atch code: %%S)# # #*A For the %Sc +,ons- in %iomedical Sciences in the University of %radford )ivision of %iomedical Sciences School of .ife Sciences University of %radford %radford )AT/ of SU%01SS1$N: *8th February * #' 2$3) 4$UNT: ''" 5ro6ect Supervisor: )r7J7%oyne

Abstract: Cancer is one of the deadly diseases in the world, claiming millions of lives. There are many reasons behind the complications which cancer putforth to the medical field.With improvement of knowledge of oncology and biotechnology, researchers have begun working on creating a novel therapeutic strategy, with minimal side effects. One of which is the usage of oncolytic viruses against cancer cells. The oncolytic viruses are able to take advantage of cancer cells strengths and break through several obstacles to infect and lyse the tumour. Moreover, with own intelligence and capabilities or with man-made modifications, the foe! which causes many deadly diseases or even cancer, can now be trusted as friend!. The once forsaken research in "#$%s, has now steadily gained confidence and funding to carry out more e&periments and even clinical trials. This article aims to introduce the rescuers! of cancer and what are the mechanisms by which they operate, inclusive of clinical trials. Introduction Cancer is one of the ma'or causes of death worldwide, accounting for about "() of all deaths worldwide. (WHO, 2013) The causes of cancer could physical, chemical or biological carcinogens. *&amples of physical carcinogens are e&cessive, harmful ultra-violet or ionising radiation. *&amples of chemical carcinogens are tobaccoor coal smoke, alcohols, aflato&ins and asbestos. *&amples of biological are parasites, bacteria and viruses. Chronic illness and genetic inheritance also contribute to tumor formation. +mongst various reasons, pathogenic, cancer-causing viruses alone give rise to estimated "") of total cases of death due to cancer. (Parkin,D.M.,2002) ,ome

e&amples of such viruses are, *pstein -arr virus causes -urkitts .ymphoma, /uman /erpes virus-0 causes 1aposi ,arcoma and cervical cancer is caused by /uman 2apilloma virus. Over the years, the medical field has fought hard against time and comple&ity of cancers, to create a conventional, prescribed course of medical treatment. The treatment consists of chemotherapy, radiotherapy, immuno-therapy, surgery or any combination. /owever, the treatment is severely challenged and becomes failure due to narrow therapeutic inde& and side effects. Moreover, treatments have very high probability of incomplete eradication of invasive primary tumor cells or dissemination of cancer cells. +s such, there is a greater need for more effective modalities. The improvement of biotechnology and understanding of oncology, scientists are thriving towards usage of virotherapy and gene therapy. +s such the oncolytic viruses are created using genetic engineering. 3figure "4

Fi ur! 1: o"!r"i!# o$ %!c&anis%s on onco'(tic "irus.)akas&i%a,H. !t a'.,(2010) Dir!ctin s(st!%ic onco'(tic "ira' d!'i"!r( to tu%ors "ia carri!r c!''s Cytokine & Growth Factor Reviews .5"354 , ""#-"56

W&at ar! onco'(tic "irus!s*

Oncolytic viruses are either naturally occuring or genetically modified viruses designed to kill cancer cells, without harming the normal cells. The viral genes act as tumor destructive agent and the capsids act as nano-si7ed nucleic acid delivery vehicle. (Waki%oto, H et al.,2002) The oncolytic viruses operate via several mechanisms which will be discussed in later sections. +(,!s o$ onco'(tic "irus!s: *nveloped8non-enveloped 9iruses which are classified as enveloped are ones that use the lipid bilayer of the cancer cells and form a coat around them. +n e&ample would be viruses from the 2o& family, such as vaccinia virus or 2aramy&ovirus family, such as Measles virus. The advantage of enveloped virus is that the glycoproteins present at the surface of virus could detect and bind to specific receptors on cancers. Two main glycoproteins are /emagglutinin and :usion proteins. The prior helps in attaching to receptors and the latter aids in fusion of virus and the host cell like cancer cells. +lso, it was noticed that enveloped viruses could utili7e larger specificity domain like that of a single-chain antibody. (-attan!o,.., 2010) /!no%!(sin '!0doub'! strand!d 1 D)A0.)A) There are various types of viral genomes. ,ome are double stranded ;<+ like adenoviruses and /erpes simple& viruses. ,ome are single stranded ;<+ viruses like 2arvoviruses. *&amples of double stranded =<+ viruses are reoviruses and single stranded =<+ are poliovirus 3positive >stranded4 and paramy&ovirus like measles virus 3negative-stranded4. )atura' 0 !n!tica''( %odi$i!d

<aturally occurring viruses mean the virus re?uires no addition of genes /!n!ra'i2!d %od! o$ actions o$ onco'(tic "irus!s @n general, there are a few ways by which the oncolytic viruses work. ,ome of the oncolytic viruses are designed or chosen to manipulate fre?uent tumour-specific mutations, some are engineered to target signalling pathways and transcriptional mechanisms that are turned-on! in tumours and some are aimed at antigens that are overe&pressed or uni?ue on tumor cells. ;espite the different mode of action, the ultimate function of the oncolytic viruses is to disrupt the cancer cells! transcriptional or translational mechanism and cause apoptosis.(Parato,3.A !t a'.,2004) Dir!ct c!'' '(sis: ;irect cell lysis is carried out by naturally occuring or selective-repllicating viruses.The aim of this mechanism is to replicate and multiply inside the cancer cell and lyse. :irstly, the viruse would adsorp to a receptor on cancer cell membrane and creates a path through to penetrate its genetic materials to the host. The genetic material would intergrate with that of the cancer cell genome. The virus then uses the cancer cell!s replicating machinery to amplify viral components. Then maturation and lysis occurs when there are surplus of viral components, many progeny are created. The new virions then produce lytic en7ymes and burst the cell or activate apoptotic pathways by generating death proteins to induce cell death. ()aka%ouri,M.!t a'., 2005) 3:igure "4

:igure "A The lytic cycle of viruses such as /,9-" or adeno viruses shows the various stages of infection. +dapted from sourceA Chiocca, +.*., 35%%54 Oncolytic viruses .Nature Reviews Cancer( 2), #(0-#$%

+n e&ample of e&periment involves /erpes ,imple& 9irus 3/,94 against murine colorectal carcinoma cells. @t was observed that there was a trigger of tumourspecific immune response and reduction in si7e. ()aka%ouri,M.!t a'., 2005)

Anti6tu%our %!c&anis%s: "4 Targetting tumour environmentsA a4 +nti-angiogenesisA +ngiogenesis is one is the hallmarks of cancer. (Hana&an.D., W!inb!r ...A.,2000).Cancer cells are able to form their own blood vessels by secreting growth factors like 9*B:. <ormally, in a mass of tumor, the inner most cells 3core region4 are deprived from o&ygen and nutrients. Therefore, such survival techni?ue is used. Oncolytic viruses are designed to trigger loss of blood flow to the core of tumor, causing depletion of o&ygen supply 3hypo&ia4 and nutrients. @nhibition of blood flow is done by blocking the synthesis of growth factors by viral genomes. .ack of blood flow would induce apoptosis in cancer cells, e&empting normal functioning cells. (-aro'in! !t a'.,2007) @n a research e&periment, vaccinia virus was used, with deletion of thymidine kinase and and its own growth factor, to reduce pathogenicity to normal cell. The virus was incorporated with soluble vascular endothelial growth factor 39*B:4 receptor >@g fusion protein 3monoclonal antibody4. to demonstrate anti-angiogenic effect 3figure 54 (/us!.3 !t a'.,2010)

:igure 5A ;iagram of how vaccinia virus perform anti-angiogenesis +dapted fromA1irn,;./. and Thorne,,./., 35%""4 Targeted and armed oncolytic po&virusesA a novel multi-mechanistic therapeutic class for cancer.

Moreover, another oncolytic virus,adeno virus has been used to inhibit angiogenesis is cancer. The adeno virus strain dl#558#CD of serotype $ and which is replicative, had deletion of C=-5 region in *"+, which binds to cellular retinoblastoma protein 3p=-4E. The deletion of *"+ C=-5 regionE in the strain was crucial in order to ensure that viral replication is only in cancerous cells. Concurrently, another strain of adeno virus +d-:lk"-:cE, which was replication-deficient, encoding soluble 9*B: receptor antibody, was used to co infect the tumour cells. The dl#558#CD strain became a helper virus and rendered its support by providing *"E functions in trans to +d-:lk"-:c strain, which increased in strength. The end result in vivo was desirable as anti-tumor and anti-angiogenic effect was observed as +d :lk-:c replicated successfully, intratumoral levels of :lk-:c proteins increased and microvessel density declined. (+&orn!.8.H., !t a'., 2009)

b4 +nti-tumoral immunity induction The immune system has two types of defense mechanism. One is non-specific or innate while the other is adaptive. The innate immune system recognises foreign particles and release "st line defense molecules like neutrophils and other pro inflammatory molecules. Then a complement cascade gets activated. On the other hand, adaptive immune system holds the postiton to eliminate pathogens in late phase of infection and creates memory cells after being triggered by presentation of antigen presenting cells 3+2C4 like macrophage and dendritic cells, from the innate immune system. One of the ma'or barriers for oncolytic viruses is to fight against effectively or evade promptly from the immune cells.under normal circumstances, despite antigens being

present on tumour epitopes, the efficiency of C;0F T cellsE was insignificant, due to strong immunosuppressive environment around tumours.(-!ru''o,:. !t a', 2010)

Oncolytic viruses are found to have the capacity to alter tumour microenvironment via oncolysis with release of molecules like tumour-associated antigen 3T++4, tumour-derived cytokine, viral nucleic acid, viral coat proteins which trigger immune response. 3 /all,1.et al,5%"54 Therefore, to activate the immune system to destroy tumour, a significantly strong firing must be released, to override the defense set by the tumour,by the oncolytic virus.

@n an e&periment, /,9-derived strains 3:us-On-/5, ,ynco-5;, -aco-"4 were used to observe antitumour immunity in neuroblastoma cells which displayed =as signalling. @t was noted that there was significant in vivo antitumour response and destruction of cells in in vitro and omission of growth of new tumors at areas further from viral-introduction site. Moreover, the strains were able to induce a strong inflammation, leading to possible maturation of dendritic cell, which is needed to act as antigen-presenting cell to signal cytoto&ic T cells. ,uch weakening of the GfirewallH created by tumour, would be favourable to the host as they would be infiltration of more immune cells to degrade tumour. (;i,H. !t a' 2007) +nother e&ample would be the e&periment done on adeno virus strain +d-;5CBMC,:, which is p"6-=b pathwayE selective and had its *" region removed and had granulocyte macrophage colony-stimulating factor 3BMC,:4 incorporated into. BMC,: aids in attracting natural killer cells to act directly on tumor and dendritic

cells which would recruit cytoto&ic T cells, signalling from lymph nodes. The tumour would be more vulnerable to the immune cells and it could be degraded. Tumour cells! immunologic tolerance was weakened as +d-;5C-BMC,: replicates within tumour and was able to present survivinE as epitope, gaining the attention of T cells. (-!ru''o,:. !t a' 2010) There is another method by which the oncolytic viruses cruise! through the sea of immune defence molecules. The usage of T-cells as cheperonesE to escort viral vectors into tumours was tested on a research. T-cells have the capability of tumour homing. Therefore, a replication-defective retroviral vector, which was incorporated with apoptotic chemokines, was able to reach its target in tumour cells. 31ottke,T.et al, 5%%04 +dding on, another interesting mode of effectively evading immune system attack and traveling through systemically to tumour site was the the clever tactic of naturally occuring virus, vaccinia, a member of 2o& virus family. @t e&ists as e&ternal-enveloped virionE3**94, whereby it would be engulfed in lipid bilayer like its host cell and would be released into the system at early stage of infection. This was seen as a great advantage because the virus was able to GpretendH it was part of the tumour as it could use the tumour!s outer lipid membrane as a cloak! and was able to infect more tumour cells. Therefore, there was increased biodistribution of virus within tumor and its microenvironment. -esides intratumoural infection, intertumoural spread was more prominent. +s such, metastatic tumours were could be reached ( 3irn,D.H.,!t a', 200<) 3figure (4.

:igure (A anti-tumoral immunity induction. ,ource adapted fromA Choi, @.1. and Iun,C.O., 35%"(4 =ecent developments in oncolytic adenovirus-based immunotherapeutic agents for use against metastatic cancers Cancer Gene Therapy 35%4, D%>D6

c4 Cytokines and signalling pathwaysA @:< pathwayA Cancers often result from defective ma'or signalling pathways which control normal cellular functions, proliferation, immune response or programmed cell death 3apoptosis4. +n e&ample of an important cascade is @nterferon 3@:<4 pathway. @nterferons 3@:<4 are versatile cytokines which activate transcription of genes whose products are mainly antiviral, anti-proliferative or immunomodulatory in function. (d! :!!r.M.= !t a'., 2001) :or e&ample, a type " cytokine, @:<-, is produced due to viral infection, as first line of defense to protect normal neighbouring cells. (;i.>. and +aink(.M.A.,2011). The @:<- increases the e&pression and transcription of @:<stimulated anti viral genes 3@,B4, such as @:<- and double stranded =<+dependent protein kinase 321=4. The @,B molecules influence the inhibition of viral genome replication and protect neighbouring cells. (8a'oura.:. !t a'.,2010) /owever in tumour, cancer cells, due to mutation of type " @:< pathway, an oncolytic virus could take advantage of the defect and replicate. @n a research, it was observed that 21=-null mice were highly susceptible to 9,9 infection. (d! :!!r.M.= !t a'., 2001) The reason being, 21= in normal cell would phosphorylate translational initiation factor e@f5a, which inhibits cellular translation and prevents host and viral protein synthesis. Jpon omission of 21=, virus could synthesi7e proteins and carry out cytolysis. (8a'oura.:. !t a'.,2010)

Moreover, in another e&periment, with respect to disrupted @:< signalling pathway, it was observed that two important @nterferon regulatory factors 3@=:4E $ and D plays a crucial role in making a cell susceptible to 9,9 infection. @n normal cell, @:< pathway has several important functions such as obstructing viral replication, activating immune cells, increasing or up-regulating antigen presentation to T cells, keeping uninfected cells safe. /owever, in cancer cells, the @:< pathway is unregulated as @=:s are silenced. Thus, anti-proliferation and cancer surveilance by the immune cells are halted. d4 +poptosis-induced oncolysisA The p$( protein is a very important protein that is the name Gguardian of the genomeH is given. (.ao, ?. !t a'., 2010) The p$( protein and a transcriptional factor, plays a crucial role in cell cycle check points and behaves as tumour suppressor, ensuring the genome is tightly conserved. Cancer cells have a defect in apoptosis mechanism, which is the reason why there is an increased cell proliferation and no programmed cell death. /owever, it was found that more than $%) of tumours e&press the wild-type p$(. Iet, the cancer cells inhibit the function of p$( by several mechanisms. (3oo,+. !t., 2011) @t has been observed that despite having a defective p$( signalling pathway, the pathway could be restored. (-&i,uk,=.@. !t a'., 2005) +n e&periment was conducted to research if oncolytic viruses could restore wildtype p$( functions in tumour cells. Tumour cells are found to have overe&pression of Mdm5 molecule which is a proto-oncoprotein and a negative regulator of p$(. +s such, p$( would be degraded, allowing cancer cells to

multiply non-stop and escape apoptosis. Thus, using replicative-competent adennovirus strains 3which were modified at either C or < terminusE which had p$( gene 3variant and wild-type4 incorporated within, tumours cells were infected. The outcome showed efficient transcriptional activity of p$( and Mdm5 inhibitory effect on p$( was observed. Therefore, with functional p$(, apoptotic molecules like -a& from -cl-5 family could be upregulated and apoptosis could occur. (3oo,+. !t., 2011) +nother e&periment done on the famous Ony&-%"$ strain of adenovirus which had *"- proteinE removed. @t was observed that irregardless of p$(-deficient or p$(mutated cancer cells, the Ony&-%"$ could infect and trigger apoptosis to occur. Clinical trialsA Clinical trials are very important part of a research for a novel therapeutic modality. The reason being, laboratory results are based on animal models, whose immune system could be compromised in order to study any pathway and behaviour of the oncolytic viruses, even when human tumor cell lines are to be used, most e&periments are done in vitro to avoid any unwanted reactions within a host. There are C main phases under clinical trial. Jnder phase ", ma&imal dose and pharmacokinetics are the focus. @n phase 5, the dose-response, type of patients, fre?uency of dose and efficacy is observed. 2hase ( is more on submission of paper works and formalities, to obtain permission from ;rug regulation authorities of the country. 2hase C would be last one, where product can be marketed. + clinical trial has to follow many rules and strict regulation as to

comply to medical ethics. There are some key factors to abide to during clinical trials.

VolunteersA The sample si7e of the number of test sub'ects to be used and drawing out eligibility of the test sub'ects is very important 3Beletneky et al.5%"54.The volunteers should not have any other medical problems that would affect the administration of the viruses and they should be well versed with the aim and procedures of the trials. They must be withdrawn upon any ma'or complications and adverse effects. Standards: Most clinical trials compare data by following the response evaluation criteria soild tumour 3=*C@,T4. The sum of biggest diameter of the target tumour is measured. +lso, partial response, complete response, progressive disease, stable disease, immune cell counts are all factors that would determine if the therapy is effective. (:an d!r :!'dt, A.A.M. !t a'., 2010)(;i, =.-. !t a'., 200<) AimA a4 2rimaryA :irstly, the safety and tolerability of the investigational medicinal product 3@M24 is important. =esearchers need to find out the ma&imum tolerance dose for each oncolytic virus therapy used. Then, the ma&imum tolerance dose is calculated. @t relates to determining whether prolonged use of the therapy would cause any

adverse effects to the mass. <ormally, an estimated ma&imum dose is given to reduce number of test sub'ects and cost of administration. b4 ,econdaryA The antitumour efficacy is monitored via tests like tumor markers, Computered Topography 3CT4 scan, cytokines assay. +lso, the survival of the test sub'ect and the period the sub'ect is free from progression or recurrance is recorded. Till date, there are several clinical trials after the re surfacing of research of oncolytic viruses. 2hase @A The first trial of 2arvOry&, a 2arvovirus /-"strain, was carried out on patients with recurrent Blioblastoma Multiforme 3B-M4 which is high-grade gliomas. The oncolytic virus was administered intravenously to study the systemic delivery and gather data to serve as platform for future trials. (/!'!tn!k( !t a'.2012) @n another trial, KL-$#C, a po& virus strain with Thymidine kinase gene removed was used against /epatitis - virus which causes /epato cellular carcinoma 3advanced refractory stage4. The po& virus is relatively well-tolerated as there is previous e&posure to most people who have encountered chicken po&! by varicella 7oster virus. The dose was in'ected intratumorally. The host cells retained thymidine kinase gene to allow replication of the oncolytic virus. The outcome of the result was satisfactory as distal tumours were also targetted despite presence of large number of antibodies. 2hase@@A

+n e&ample of phase @@ trial consists of replication-competent adenovirus strain, Ony&-%"$. @t was tested against s?uamous cell carcinoma of head and neck 3,CC/<4. @t was observed the virus remains in the blood stream for only a short time and some discomforts of pain at site of in'ection and mild fever was felt by the test sub'ects. ()!%uaitis,=. !t a'., 2001) 2hase @@@A =eolysin is commercial product of Oncolytics -iotech, is a name for reovirus strain, in combination with two drugs, 2aclita&el and Carboplatin. -eing able to clear phase @@ would mean that the therapy was efficient in reducing tumour si7e and ma&imal dose has been determined. ()!t r!sourc!s int!rnationa', 2012) +dding on to the list of victory, Onco9e& BM-C,:by -io9e& company, a strain of /,9-" with genetic modification, has also progressed to phase @@@. @n phase @@, the tumour si7e reduced to more than $%) 3melanoma4. Onco9e&BM-C,: was able to infect cancer cells and replicate and lyse, recruiting immune cells to clear out the debris. +s a strong oncolytic virus, it has shown remarkable results in breast , pancreatic, head and neck cancer and melanoma. With no modifications and tests, the strain stands a great chance to tackle more cancer types. (?io:!A Inc, 2012)

Conclusion There are several types of oncolytic viruses with various types of mechanisms by which they function. /owever, there are several drawbacks and contradictions and problems laying which each e&periment done. :or instance, there could transient

viremia, where by the oncolytic virus is removed from blood stream ?uickly before it could e&ert its impact or there could incomplete incorporation of viral genome with the cancer cell, failing to turn on apoptosis . ;espite these drawbacks, there is hope that oncolytic viruses would be a novel therapeutic strategy earlier than once e&pected.

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.iu, T.C., /wang, T., 2ark, -./., -ell, K and 1irn, ;./. 35%%04. The Targeted Oncolytic 2o&virus KL-$#C ;emonstrates +ntitumoral, +ntivascular, and +nti-/-9 +ctivities in 2atients With /epatocellular Carcinoma. Molecular Therapy 1("4, "6(D>"6C5. Marcato, 2., ;ean, C., Biacomantonio, C.+., and .ee, 2.W.1. 35%%#4. Oncolytic =eovirus *ffectively Targets -reast Cancer ,tem Cells. American Society o/ Gene Therapy )(1#$")', ")". <akamori, M., :u, L., =ousseau, =., Chen, ,.I., and Ohang, L. 35%%C4. ;estruction of nonimmunogenic mammary tumor cells by a fusogenic oncolytic herpes simple& virus induces potent antitumor immunity. Molecular Therapy "(!#$ 1!0,11! )akas&i%a,H. !t a'.,(2010) ;irecting systemic oncolytic viral delivery to tumors via carrier cells Cytokine & Growth Factor Reviews .21(24 , ""#-"56 )!%unaitis, =., 3&uri, F., /an'(, I., Ars!n!au, =., Posn!r, M., :ok!s, @., 3u&n, =., McCarty. T., ;and!rs, 8., ?'ackburn, A., .o%!', ;., .and'!", ?., 3a(!, 8., and 3irn. D,. 35%%04. 2hase @@ trial of intratumoral administration of O<IL-%"$, a replicationselective adenovirus, in patients with refractory head and neck cancer. Gene Therapy 1(&#$ (D6-(05. <emunaitis, K., 1huri, :., Banly, @., +rseneau, K., 2osner, M., 9okes, *., 1uhn, K., McCarty, T., .anders, ,., -lackburn, +., =omel, .., =andlev, -., 1aye, ,., 1irn, ;. 35%"%4. 2hase @@ trial of intratumoral administration of O<IL-%"$, a replication-selective adenovirus, in patients with refractory head and neck cancer. .ournal o/ Clinical 9ncolo%y !( "#$ 50#-5#0. 2restwich, =.K., *rrington, :., ,teele, ..2. ilett, *.K., Morgan, =.,.M., /arrington, 1.K., 2andha, /.,., ,elby, 2.K., 9ile, =.B., and Melcher, +.+. 35%%#4. *valuation of an +ttenuated 9esicular ,tomatitis =eciprocal /uman ;endritic Cell><atural 1iller Cell @nteractions @nduce +ntitumor +ctivity :ollowing Tumor Cell @nfection by Oncolytic =eovirus. The .ournal o/ 7mmunolo%y 0&, C("5>C(5". 2restwich, =.K., @lett, *.K., *rringtton, :., ;ia7, =.M., ,teele, ..2., Ottke, T.1., Thompson, K., Balivo, :., /arrington, 1., 2andha, /., ,elby, 2., 9ile, =., and Melcher, +. 35%%#4.@mmuneMediated +ntitumor+ctivity of =eovirus @s =e?uired for Therapy and @s @ndependent of ;irect 9iral Oncolysis and =eplication. Clinical Cancer Research !( &#$ C("5-C(5". 2restwich,=.K.,*rrington,:., ,teele,..2.,@lett,*.K., Morgan,=.,.M.,/arrington,1.K., 2andha,/.,.,,elby,2.K.,=ichard B. 9ile,and +lan +. Melcher(E ,aloura, 9., Wang, ..C.,, :ridlender, O.B., ,un, K., Chang, B., 1apoor, 9., ,terman, ;./., /arty, =.<., Okumura, +., -arber, B .<., 9ile, =.B., :ederspiel, M.K., =ussell, ,.K., .it7ky, .., and +lbelda, ,.M. 35%"%4. 9irus 9ector *&pressing @nterferon-b for Jse in Malignant 2leural MesotheliomaA /eterogeneity in @nterferon =esponsiveness ;efines 2otential *fficacy. 3uman Gene Therapy ' $ $"-6C.

=ao, -., .eeuwen, @.M.M.9., /iggins, M., Campbell, K., Thompson, +.M., .ane, ;.2., and .ain, ,. 35%"%4. *valuation of an +ctinomycin ;89L-60% aurora kinase inhibitor combination in p$(-based cyclotherapy. Oncotarget "3D4, 6(#-6$%. =ogulski, 1.=., :reytag, ,.O., Ohang, 1., Bilbert, K.;., 2aielli, ;..., 1im, K./., /eise, C.C., and 1im, ;./. 35%%%4. @n 9ivo +ntitumor +ctivity of O<IL-%"$ @s @nfluenced by p$( ,tatus and @s +ugmented by =adiotherapy. Cancer Research 1*$ ""#(-""#6. ,obol, 2., -oudreau, K., ,tephenson, 1., Wan, I., .itchy, -., and Mossman, 1. 35%""4. +daptive +ntiviral @mmunity @s a ;eterminant of the Therapeutic ,uccess of Oncolytic 9irotherapy. Molecular Therapy "# 354, (($>(CC. +&orn!, 8H., +a%, ?.C, 3irn, D.H., -onta , -.H., and 3uo, -.=. 35%%64. ,elective intratumoral amplification of an antiangiogenic vector by an oncolytic virus produces enhanced antivascular and anti-tumor efficacy. Molecular Therapy &(!#, #(0-#C6. 9an der 9eldt, +.+.M., Mei'erink, M.=., van der *artwegh, +.K.M., /aanen, K.-.+.B., and -oven, *. 35%"%4. Choi response criteria for early prediction of clinical outcome in patients with metastatic renal cell cancer treated with sunitinib. -ritish .ournal o/ Cancer *', 0%(> 0%#.

9eer, M.K., /olko, M., :revel, M., Walker, *., ;er, ,., 2aran'ape, K.M., ,ilverman, =./., and Wiliiams -.=.B. 35%%"4. :unctional classification of interferon-stimulated genes identified using microarrays. .ournal o/ 2eu:ocyte -iolo%y 1"$ #"5-#5%. ,pilker, ,., "#0C. Buide to Clinical Trials. =aven 2ress PonlineQ. +vailable atA RhttpA88www.virginia.edu8vpr8irb8/,=Sdocs8C.@<@C+.ST=@+.,S2hases.pdfT P+ccessed 5Dth :ebruary 5%"(Q. Wang, I., Tian, 2.K., and Wei, K. .ytic cycleA a defining process in oncolytic 9irotherapy. PonlineQ. +vailable atA RhttpA88www.math.wm.edu8U'ptian8preprints8pr-#-dodevorith.pdfT P+ccessed 5%th :ebruary 5%"(Q -io9e& Commences Onco9*L 3BM-C,:4 2hase ( Trial in Metastatic Melanoma. PonlineQ. +vailable atA R&tt,:00###.,rn!#s#ir!.co%0n!#s6r!'!as!s0bio"!A6co%%!nc!s6 onco"!A6 %6cs$6,&as!636tria'6in6%!tastatic6%!'ano%a691<045D2.&t%'T P+ccessed ;ateA ""th Kanuary 5%"(Q

5%"5. Oncolytics reports early positive data from 2hase @@@ study of =eolysin. PonlineQ. +vailable atA RhttpA88www.drugdevelopment-technology.com8news8newsoncolytics-reportsearly-positive-data-from-phase-iii-study-of-reolysinT P+ccessed ;ateA 5"st Kanuary 5%"(Q ,pilker, ,., "#0C. Buide to Clinical Trials. =aven 2ress PonlineQ. +vailable atA RhttpA88www.virginia.edu8vpr8irb8/,=Sdocs8C.@<@C+.ST=@+.,S2hases.pdfT P+ccessed 5Dth :ebruary 5%"(Q.