title: Tissue inhibitor of metalloproteinase-3 expression from an oncolytic adenovirus inhibits matrix metalloproteinase activity in vivo without affecting antitumor efficacy in malignant glioma.
abstract: Oncolytic adenoviruses exhibiting tumor-selective replication are promising anticancer agents. Insertion and expression of a transgene encoding tissue inhibitor of metalloproteinase-3 (TIMP-3), which has been reported to inhibit angiogenesis and tumor cell infiltration and induce apoptosis, may improve the antitumor activity of these agents. To assess the effects of TIMP-3 gene transfer to glioma cells, a replication-defective adenovirus encoding TIMP-3 (Ad.TIMP-3) was employed. Ad.TIMP-3 infection of a panel of glioma cell cultures decreased the proliferative capacity of these cells and induced morphologic changes characteristic for apoptosis. Next, a conditionally replicating adenovirus encoding TIMP-3 was constructed by inserting the TIMP-3 expression cassette into the E3 region of the adenoviral backbone containing a 24-bp deletion in E1A. This novel oncolytic adenovirus, AdDelta24TIMP-3, showed enhanced oncolytic activity on a panel of primary cell cultures and two glioma cell lines compared with the control oncolytic virus AdDelta24Luc. In vivo inhibition of matrix metalloproteinase (MMP) activity by AdDelta24TIMP-3 was shown in s.c. glioma xenografts. The functional activity of TIMP-3 was imaged noninvasively using a near-IR fluorescent MMP-2-activated probe. Tumoral MMP-2 activity was significantly reduced by 58% in the AdDelta24TIMP-3-treated tumors 24 hours after infection. A study into the therapeutic effects of combined oncolytic and antiproteolytic therapy was done in both a s.c. and an intracranial model for malignant glioma. Treatment of s.c. (U-87MG) or intracranial (U-87deltaEGFR) tumors with AdDelta24TIMP-3 and AdDelta24Luc both significantly inhibited tumor growth and prolonged survival compared with PBS-treated controls. However, expression of TIMP-3 in the context of AdDelta24 did not significantly affect the antitumor efficacy of this oncolytic agent.
authors.affiliation: Department of Neurosurgery, Division of Gene Therapy, VU University Medical Center, Amsterdam, The Netherlands. M.Lamfers@vumc.nl
authors.forename: Martine L M
authors.initials: ML
authors.lastname: Lamfers
authors.forename: Davide
authors.initials: D
authors.lastname: Gianni
authors.forename: Ching-Hsuan
authors.initials: CH
authors.lastname: Tung
authors.forename: Sander
authors.initials: S
authors.lastname: Idema
authors.forename: Frederik H E
authors.initials: FH
authors.lastname: Schagen
authors.forename: Jan E
authors.initials: JE
authors.lastname: Carette
authors.forename: Paul H A
authors.initials: PH
authors.lastname: Quax
authors.forename: Victor W
authors.initials: VW
authors.lastname: Van Beusechem
authors.forename: W Peter
authors.initials: WP
authors.lastname: Vandertop
authors.forename: Clemens M F
authors.initials: CM
authors.lastname: Dirven
authors.forename: E Antonio
authors.initials: EA
authors.lastname: Chiocca
authors.forename: Winald R
authors.initials: WR
authors.lastname: Gerritsen
mesh_terms: D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D000818:Animals; D048708:Cell Growth Processes / Q000235:genetics; D045744:Cell Line, Tumor; D018014:Gene Transfer Techniques; D005910:Glioma / Q000201:enzymology / Q000235:genetics / Q000628:therapy* / Q000821:virology*; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D061965:Matrix Metalloproteinase Inhibitors; D051379:Mice; D008819:Mice, Nude; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication; D023041:Xenograft Model Antitumor Assays
chemical_list: D061965:Matrix Metalloproteinase Inhibitors; D019717:Tissue Inhibitor of Metalloproteinase-3; D020778:Matrix Metalloproteinase 2