Evaluation of the Safety and Biodistribution of M032,an Attenuated Herpes Simplex Virus Type 1 Expressing hIL-12,After Intracerebral Administration to Aotus Nonhuman Primates
Restricted accessResearch articleFirst published online March, 2014
Evaluation of the Safety and Biodistribution of M032,an Attenuated Herpes Simplex Virus Type 1 Expressing hIL-12,After Intracerebral Administration to Aotus Nonhuman Primates
Herpes simplex virus type 1 (HSV-1) mutants lacking the γ134.5 neurovirulence loci are promising agents for treating malignant glioma. Arming oncolytic HSV-1 to express immunostimulatory genes may potentiate therapeutic efficacy. We have previously demonstrated improved preclinical efficacy, biodistribution, and safety of M002, a γ134.5-deleted HSV-1 engineered to express murine IL-12. Herein, we describe the safety and biodistribution of M032, a γ134.5-deleted HSV-1 virus that expresses human IL-12 after intracerebral administration to nonhuman primates, Aotus nancymae. Cohorts were administered vehicle, 106, or 108 pfu of M032 on day 1 and subjected to detailed clinical observations performed serially over a 92-day trial. Animals were sacrificed on days 3, 31, and 91 for detailed histopathologic assessments of all organs and to isolate and quantify virus in all organs. With the possible exception of one animal euthanized on day 16, neither adverse clinical signs nor sex- or dose-related differences were attributed to M032. Elevated white blood cell and neutrophil counts were observed in virus-injected groups on day 3, but no other significant changes were noted in clinical chemistry or coagulation parameters. Minimal to mild inflammation and fibrosis detected, primarily in meningeal tissues, in M032-injected animals on days 3 and 31 had mostly resolved by day 91. The highest viral DNA levels were detected at the injection site and motor cortex on day 3 but decreased in central nervous system tissues over time. These data demonstrate the requisite safety of intracerebral M032 administration for consideration as a therapeutic for treating malignant brain tumors.
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References
1.
AghiM., ChouT.C., SulingK., et al. (1999). Multimodal cancer treatment mediated by a replicating oncolytic virus that delivers the oxazaphosphorine/rat cytochrome P450 2B1 and ganciclovir/herpes simplex virus thymidine kinase gene therapies. Cancer Res., 59, 3861–3865.
2.
AlbiniA., BrigatiC., VenturaA., et al. (2009). Angiostatin anti-angiogenesis requires IL-12: the innate immune system as a key target. J. Transl. Med., 7, 5.
3.
AndreanskyS., HeB., Van CottJ., et al. (1998). Treatment of intracranial gliomas in immunocompetent mice using herpes simplex viruses that express murine interleukins. Gene Ther., 5, 121–130.
4.
BarahonaH., MelendezL.V., HuntR.D., and DanielM.D. (1976). The owl monkey (Aotus trivirgatus) as an animal model for viral diseases and oncologic studies. Lab. Anim. Sci., 26, 1104–1112.
5.
CassadyK.A., and ParkerJ.N. (2010). Herpesvirus vectors for therapy of brain tumors. Open. Virol. J, 4, 103–108.
6.
ChambersR., GillespieG.Y., SoroceanuL., et al. (1995). Comparison of genetically engineered herpes simplex viruses for the treatment of brain tumors in a scid mouse model of human malignant glioma. Proc. Natl. Acad. Sci. USA, 92, 1411–1415.
7.
ChaseM., ChungR.Y., and ChioccaE.A. (1998). An oncolytic viral mutant that delivers the CYP2B1 transgene and augments cyclophosphamide chemotherapy. Nat. Biotechnol., 16, 444–448.
8.
ChouJ., KernE.R., WhitleyR.J., and RoizmanB. (1990). Mapping of herpes simplex virus-1 neurovirulence to gamma 134.5, a gene nonessential for growth in culture. Science, 250, 1262–1266.
9.
FarassatiF., YangA.D., and LeeP.W. (2001). Oncogenes in Ras signalling pathway dictate host-cell permissiveness to herpes simplex virus 1. Nat. Cell Biol., 3, 745–750.
10.
FeingoldK.R., and GrunfeldC. (2010). The acute phase response inhibits reverse cholesterol transport. J. Lipid Res., 51, 682–684.
11.
GoodwinJ.M., SchmittA.D., McginnC.M., et al. (2012). Angiogenesis inhibition using an oncolytic herpes simplex virus expressing endostatin in a murine lung cancer model. Cancer Invest., 30, 243–250.
12.
GozaloA., and MontoyaE. (1990). Mortality causes of owl monkeys (Aotus nancymae and Aotus vociferans) in captivity. J. Med. Primatol., 19, 69–72.
13.
GuffeyM.B., ParkerJ.N., LuckettW.S.Jr., et al. (2007). Engineered herpes simplex virus expressing bacterial cytosine deaminase for experimental therapy of brain tumors. Cancer Gene Ther., 14, 45–56.
14.
HanZ.Q., AssenbergM., LiuB.L., et al. (2007). Development of a second-generation oncolytic Herpes simplex virus expressing TNFalpha for cancer therapy. J. Gene Med., 9, 99–106.
15.
HarrowS., PapanastassiouV., HarlandJ., et al. (2004). HSV1716 injection into the brain adjacent to tumour following surgical resection of high-grade glioma: safety data and long-term survival. Gene Ther., 11, 1648–1658.
16.
HeB., GrossM., and RoizmanB. (1997). The gamma(1)34.5 protein of herpes simplex virus 1 complexes with protein phosphatase 1alpha to dephosphorylate the alpha subunit of the eukaryotic translation initiation factor 2 and preclude the shutoff of protein synthesis by double-stranded RNA-activated protein kinase. Proc. Natl. Acad. Sci. USA, 94, 843–848.
17.
HeB., GrossM., and RoizmanB. (1998). The gamma134.5 protein of herpes simplex virus 1 has the structural and functional attributes of a protein phosphatase 1 regulatory subunit and is present in a high molecular weight complex with the enzyme in infected cells. J. Biol. Chem., 273, 20737–20743.
18.
HellumsE.K., MarkertJ.M., ParkerJ.N., et al. (2005). Increased efficacy of an interleukin-12-secreting herpes simplex virus in a syngeneic intracranial murine glioma model. Neuro Oncol., 7, 213–224.
19.
HunterW.D., MartuzaR.L., FeigenbaumF., et al. (1999). Attenuated, replication-competent herpes simplex virus type 1 mutant G207: safety evaluation of intracerebral injection in nonhuman primates. J. Virol., 73, 6319–6326.
20.
KatzinD.S., ConnorJ.D., WilsonL.A., and SextonR.S. (1967). Experimental herpes simplex infection in the owl monkey. Proc. Soc. Exp. Biol. Med., 125, 391–398.
21.
KennedyP.G., AdamsJ.H., GrahamD.I., and ClementsG.B. (1988). A clinico-pathological study of herpes simplex encephalitis. Neuropathol. Appl. Neurobiol., 14, 395–415.
22.
KhovidhunkitW., KimM.S., MemonR.A., et al. (2004). Effects of infection and inflammation on lipid and lipoprotein metabolism: mechanisms and consequences to the host. J. Lipid Res., 45, 1169–1196.
23.
KochF., StanzlU., JenneweinP., et al. (1996). High level IL-12 production by murine dendritic cells: upregulation via MHC class II and CD40 molecules and downregulation by IL-4 and IL-10. J. Exp. Med., 184, 741–746.
24.
KrammC.M., ChaseM., HerrlingerU., et al. (1997). Therapeutic efficiency and safety of a second-generation replication-conditional HSV1 vector for brain tumor gene therapy. Hum. Gene Ther., 8, 2057–2068.
25.
LiuB.L., RobinsonM., HanZ.Q., et al. (2003). ICP34.5 deleted herpes simplex virus with enhanced oncolytic, immune stimulating, and anti-tumour properties. Gene Ther., 10, 292–303.
26.
LiuT.C., ZhangT., FukuharaH., et al. (2006). Oncolytic HSV armed with platelet factor 4, an antiangiogenic agent, shows enhanced efficacy. Mol. Ther., 14, 789–797.
27.
LowenstineL.J. (2003). A primer of primate pathology: lesions and nonlesions. Toxicol. Pathol., 31Suppl, 92–102.
28.
ManettiR., GerosaF., GiudiziM.G., et al. (1994). Interleukin 12 induces stable priming for interferon gamma (IFN-gamma) production during differentiation of human T helper (Th) cells and transient IFN-gamma production in established Th2 cell clones. J. Exp. Med., 179, 1273–1283.
29.
MarkertJ.M., MalickA., CoenD.M., and MartuzaR.L. (1993). Reduction and elimination of encephalitis in an experimental glioma therapy model with attenuated herpes simplex mutants that retain susceptibility to acyclovir. Neurosurgery, 32, 597–603.
30.
MarkertJ.M., MedlockM.D., RabkinS.D., et al. (2000). Conditionally replicating herpes simplex virus mutant, G207 for the treatment of malignant glioma: results of a phase I trial. Gene Ther., 7, 867–874.
31.
MarkertJ.M., LiechtyP.G., WangW., et al. (2009). Phase Ib trial of mutant herpes simplex virus G207 inoculated pre-and post-tumor resection for recurrent GBM. Mol. Ther., 17, 199–207.
32.
MarkertJ.M., CodyJ.J., ParkerJ.N., et al. (2012). Preclinical evaluation of a genetically engineered herpes simplex virus expressing interleukin-12. J. Virol., 86, 5304–5313.
33.
MartuzaR.L., MalickA., MarkertJ.M., et al. (1991). Experimental therapy of human glioma by means of a genetically engineered virus mutant. Science, 252, 854–856.
34.
MeignierB., MartinB., WhitleyR.J., and RoizmanB. (1990). In vivo behavior of genetically engineered herpes simplex viruses R7017 and R7020. II. Studies in immunocompetent and immunosuppressed owl monkeys (Aotus trivirgatus). J. Infect. Dis., 162, 313–321.
35.
MinetaT., RabkinS.D., YazakiT., et al. (1995). Attenuated multi-mutated herpes simplex virus-1 for the treatment of malignant gliomas. Nat. Med., 1, 938–943.
36.
MurphyW.J., WelniakL., BackT., et al. (2003). Synergistic anti-tumor responses after administration of agonistic antibodies to CD40 and IL-2: coordination of dendritic and CD8+ cell responses. J. Immunol., 170, 2727–2733.
37.
NakamoriM., FuX., PettawayC.A., and ZhangX. (2004). Potent antitumor activity after systemic delivery of a doubly fusogenic oncolytic herpes simplex virus against metastatic prostate cancer. Prostate, 60, 53–60.
38.
NakamuraH., MullenJ.T., ChandrasekharS., et al. (2001). Multimodality therapy with a replication-conditional herpes simplex virus 1 mutant that expresses yeast cytosine deaminase for intratumoral conversion of 5-fluorocytosine to 5-fluorouracil. Cancer Res., 61, 5447–5452.
39.
PapanastassiouV., RamplingR., FraserM., et al. (2002). The potential for efficacy of the modified (ICP 34.5(−)) herpes simplex virus HSV1716 following intratumoural injection into human malignant glioma: a proof of principle study. Gene Ther., 9, 398–406.
40.
ParkerJ.N., GillespieG.Y., LoveC.E., et al. (2000). Engineered herpes simplex virus expressing IL-12 in the treatment of experimental murine brain tumors. Proc. Natl. Acad. Sci. USA, 97, 2208–2213.
41.
ParkerJ.N., MelethS., HughesK.B., et al. (2005). Enhanced inhibition of syngeneic murine tumors by combinatorial therapy with genetically engineered HSV-1 expressing CCL2 and IL-12. Cancer Gene Ther., 12, 359–368.
42.
ParkerJ.N., BauerD.F., CodyJ.J., and MarkertJ.M. (2009). Oncolytic viral therapy of malignant glioma. Neurotherapeutics, 6, 558–569.
43.
PrabhakarS., BrennerG.J., SungB., et al. (2010). Imaging and therapy of experimental schwannomas using HSV amplicon vector-encoding apoptotic protein under Schwann cell promoter. Cancer Gene Ther., 17, 266–274.
44.
PreusserM., De RibaupierreS., WohrerA., et al. (2011). Current concepts and management of glioblastoma. Ann. Neurol., 70, 9–21.
45.
PylesR.B., WarnickR.E., ChalkC.L., et al. (1997). A novel multiply-mutated HSV-1 strain for the treatment of human brain tumors. Hum. Gene Ther., 8, 533–544.
46.
RamplingR., CruickshankG., PapanastassiouV., et al. (2000). Toxicity evaluation of replication-competent herpes simplex virus (ICP 34.5 null mutant 1716) in patients with recurrent malignant glioma. Gene Ther., 7, 859–866.
47.
SgadariC., AngiolilloA.L., and TosatoG. (1996). Inhibition of angiogenesis by interleukin-12 is mediated by the interferon-inducible protein 10. Blood, 87, 3877–3882.
48.
ShahA.C., BenosD., GillespieG.Y., and MarkertJ.M. (2003). Oncolytic viruses: clinical applications as vectors for the treatment of malignant gliomas. J. Neurooncol., 65, 203–226.
49.
SimpsonG.R., and CoffinR.S. (2009). Construction and characterization of an oncolytic HSV vector containing a fusogenic glycoprotein and prodrug activation for enhanced local tumor control. Methods Mol. Biol., 542, 551–564.
50.
SmithK.D., MezhirJ.J., BickenbachK., et al. (2006). Activated MEK suppresses activation of PKR and enables efficient replication and in vivo oncolysis by Deltagamma(1)34.5 mutants of herpes simplex virus 1. J. Virol., 80, 1110–1120.
51.
StewartV.A. (2003). Plasmodium vivax under the microscope: the Aotus model. Trends Parasitol., 19, 589–594.
52.
StuppR., HegiM.E., MasonW.P., et al. (2009). Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase III study: 5-year analysis of the EORTC-NCIC trial. Lancet Oncol., 10, 459–466.
53.
SuzukiS., UmezuY., SaijoY., et al. (1998). Exogenous recombinant human IL-12 augments MHC class I antigen expression on human cancer cells in vitro. Tohoku J. Exp. Med., 185, 223–226.
54.
TakaokaH., TakahashiG., OgawaF., et al. (2011). A novel fusogenic herpes simplex virus for oncolytic virotherapy of squamous cell carcinoma. Virol. J., 8, 294.
55.
TodoT., FeigenbaumF., RabkinS.D., et al. (2000). Viral shedding and biodistribution of G207, a multimutated, conditionally replicating herpes simplex virus type 1, after intracerebral inoculation in Aotus. Mol. Ther., 2, 588–595.
56.
TrinchieriG. (2003). Interleukin-12 and the regulation of innate resistance and adaptive immunity. Nat. Rev. Immunol., 3, 133–146.
57.
VargheseS., NewsomeJ.T., RabkinS.D., et al. (2001). Preclinical safety evaluation of G207, a replication-competent herpes simplex virus type 1, inoculated intraprostatically in mice and nonhuman primates. Hum. Gene Ther., 12, 999–1010.
58.
VoestE.E., KenyonB.M., O'ReillyM.S., et al. (1995). Inhibition of angiogenesis in vivo by interleukin 12. J. Natl. Cancer Inst., 87, 581–586.
59.
WalkerJ.D., SehgalI., and KousoulasK.G. (2011). Oncolytic herpes simplex virus 1 encoding 15-prostaglandin dehydrogenase mitigates immune suppression and reduces ectopic primary and metastatic breast cancer in mice. J. Virol., 85, 7363–7371.
60.
WiggintonJ.M., LeeJ.K., WiltroutT.A., et al. (2002). Synergistic engagement of an ineffective endogenous anti-tumor immune response and induction of IFN-gamma and Fas-ligand-dependent tumor eradication by combined administration of IL-18 and IL-2. J. Immunol., 169, 4467–4474.
61.
YooJ.Y., HaseleyA., BrataszA., et al. (2012). Antitumor efficacy of 34.5ENVE: a transcriptionally retargeted and “Vstat120”-expressing oncolytic virus. Mol. Ther., 20, 287–297.
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