Synergistic Antitumor Effect of Recombinant Adeno-Associated Virus-Mediated Pigment Epithelium-Derived Factor with Hyperthermia on Solid Tumor

Abstract

Adeno-associated virus (AAV) is an ideal choice for gene delivery; however, its further development has been limited owing to its low transduction efficiency. DNA-damaging agents can improve AAV-mediated transgene expression. Hyperthermia, as one of the oldest documented tumor treatment modalities, can cause DNA damage as well. However, combined treatment consisting of hyperthermia and AAV-mediated gene therapy has not been reported yet. In this work we investigated whether therapy consisting of AAV-mediated pigment epithelium-derived factor (PEDF) delivery combined with hyperthermia has synergistic antitumor effect on established solid tumors. We produced the recombinant AAV encoding PEDF (rAAV-PEDF). The therapeutic effect of rAAV-PEDF plus hyperthermia was evaluated in a subcutaneous fibrosarcoma mouse model, and the possible mechanism of antitumor effect was investigated. We found that rAAV-PEDF could infect a murine fibrosarcoma cell line (Meth-A) and express PEDF protein with bioactivity in vitro. In addition, in vivo experiments suggested that the combination of rAAV-PEDF with hyperthermia could significantly suppress tumor growth and prolong survival time of treated mice. Immunofluorescence studies indicated that the combination therapy could inhibit angiogenesis and induce apoptosis in tumor tissues. An immunohistochemistry assay of tumor tissue showed that PEDF expression in the combined treatment group was significantly higher than in the rAAV-PEDF group, which implied that hyperthermia could improve the expression of PEDF protein in vivo. No significant differences were observed in each group by hematoxylin–eosin staining of major organs, serum chemistry test, and complete blood assay. These results indicate that the combination of rAAV-PEDF with hyperthermia has synergistic therapeutic effects on established solid tumors, with no side effects. In addition, hyperthermia could improve AAV-mediated transgene expression, which suggests that hyperthermia could serve as a promising adjunctive therapy for AAV-mediated gene therapy.

Get full access to this article

View all access options for this article.

References

Abdollahi

, and Folkman

(2010). Evading tumor evasion: Current concepts and perspectives of anti-angiogenic cancer therapy. Drug Resist. Updat., 13, 16.

Abdollahi

, Lipson

K.E.

, Sckell

, et al. (2003). Combined therapy with direct and indirect angiogenesis inhibition results in enhanced antiangiogenic and antitumor effects. Cancer Res., 63, 8890–8898.

Alexander

I.E.

, Russell

D.W.

, and Miller

(1994). DNA-damaging agents greatly increase the transduction of nondividing cells by adeno-associated virus vectors. J. Virol., 68, 8282–8287.

Bettaieb

, Wrzal

P.K.

, and Averill-Bates

D.A.

(2013). Hyperthermia: Cancer treatment and beyond. In Cancer Treatment-Conventional and Innovative Approaches. Rangel

, ed. (InTech Publishers, Rijeka, Croatia) pp. 257–283.

Blouin

, Brument

, Toublanc

, et al. (2004). Improving rAAV production and purification: Towards the definition of a scaleable process. J. Gene Med., 6, S223–S228.

Chen

, Yang

L.-l.

, Yang

H.-s.

, et al. (2008). Synergistic antitumor effect of CXCL10 with hyperthermia. J. Cancer Res. Clin. Oncol., 134, 679–687.

Chung

D.C.

, Fogelgren

, Park

K.M.

, et al. (2011). Adeno-associated virus-mediated gene transfer to renal tubule cells via a retrograde ureteral approach. Nephron Extra, 1, 217–223.

E.T.

, Dass

C.R.

, and Choong

P.F.

(2006). Pigment epithelium-derived factor: A multimodal tumor inhibitor. Mol. Cancer Ther., 5, 1641–1646.

Eppink

, Krawczyk

P.M.

, Stap

, et al. (2012). Hyperthermia-induced DNA repair deficiency suggests novel therapeutic anti-cancer strategies. Int. J. Hyperthermia, 28, 509–517.

10.

Ferrari

F.K.

, Samulski

, Shenk

, et al. (1996). Second-strand synthesis is a rate-limiting step for efficient transduction by recombinant adeno-associated virus vectors. J. Virol., 70, 3227–3234.

11.

Fischer

A.H.

, Jacobson

K.A.

, Rose

, et al. (2008). Hematoxylin and eosin staining of tissue and cell sections. CSH Protoc., 2008, pdb.prot4986.

12.

Fisher

K.J.

, Gao

G.-P.

, Weitzman

M.D.

, et al. (1996). Transduction with recombinant adeno-associated virus for gene therapy is limited by leading-strand synthesis. J. Virol., 70, 520–532.

13.

Folkman

(1971). Tumor angiogenesis: Therapeutic implications. N. Engl. J. Med., 285, 1182–1186.

14.

Grieger

J.C.

, Choi

V.W.

, and Samulski

R.J.

(2006). Production and characterization of adeno-associated viral vectors. Nat. Protocols, 1, 1412–1428.

15.

Hanahan

, and Folkman

(1996). Patterns and emerging mechanisms of the angiogenic switch during tumorigenesis. Cell, 86, 353–364.

16.

Jaffe

E.A.

, Nachman

R.L.

, Becker

C.G.

, et al. (1973). Culture of human endothelial cells derived from umbilical veins: Identification by morphologic and immunologic criteria. J. Clin. Invest., 52, 2745–2756.

17.

Jiang

, Liu

, Xiang

, et al. (2011). Synergistic antitumor effect of AAV-mediated TRAIL expression combined with cisplatin on head and neck squamous cell carcinoma. BMC Cancer, 11, 54.

18.

Kay

M.A.

, Glorioso

J.C.

, and Naldini

(2001). Viral vectors for gene therapy: The art of turning infectious agents into vehicles of therapeutics. Nat. Med., 7, 33–40.

19.

, Tian

, Hou

J.-m.

, et al. (2005). Improved therapeutic effectiveness by combining recombinant CXC chemokine ligand 10 with Cisplatin in solid tumors. Clin. Cancer Res., 11, 4217–4224.

20.

, and Huang

(2000). Nonviral gene therapy: Promises and challenges. Gene Ther., 7, 31–34.

21.

, Lin

, Chiang

, et al. (2002). Intratumoral gene therapy of malignant brain tumor in a rat model with angiostatin delivered by adeno-associated viral (AAV) vector. Gene Ther., 9, 2–11.

22.

Moss

R.B.

, Milla

, Colombo

, et al. (2007). Repeated aerosolized AAV-CFTR for treatment of cystic fibrosis: A randomized placebo-controlled phase 2B trial. Hum. Gene Ther., 18, 726–732.

23.

Pang

, Yi

, et al. (2009). Morelloflavone, a biflavonoid, inhibits tumor angiogenesis by targeting rho GTPases and extracellular signal-regulated kinase signaling pathways. Cancer Res., 69, 518–525.

24.

Patan

(2004). Vasculogenesis and angiogenesis. Cancer Treat. Res., 117, 3–32.

25.

Pennock

G.K.

, Waterfield

, and Wolchok

J.D.

(2012). Patient responses to ipilimumab, a novel immunopotentiator for metastatic melanoma: How different are these from conventional treatment responses?. Am. J. Clin. Oncol., 35, 606–611.

26.

Ponnazhagan

, Curiel

D.T.

, Shaw

D.R.

, et al. (2001). Adeno-associated virus for cancer gene therapy. Cancer Res., 61, 6313–6321.

27.

Roca

, Primo

, Valdembri

, et al. (2003). Hyperthermia inhibits angiogenesis by a plasminogen activator inhibitor 1-dependent mechanism. Cancer Res., 63, 1500–1507.

28.

Samulski

, Zhu

, Xiao

, et al. (1991). Targeted integration of adeno-associated virus (AAV) into human chromosome 19. EMBO J., 10, 3941–3950. [Published erratum in EMBO J. 11:1228.]

29.

Song

C.W.

(1984). Effect of local hyperthermia on blood flow and microenvironment: A review. Cancer Res., 44, 4721s.

30.

Speit

, and Rothfuss

(2012). The comet assay: A sensitive genotoxicity test for the detection of DNA damage and repair. In DNA Repair Protocols (Springer, New York) pp. 79–90.

31.

Tombran-Tink

, and Barnstable

C.J.

(2003a). PEDF: A multifaceted neurotrophic factor. Nat. Rev. Neurosci., 4, 628–636.

32.

Tombran-Tink

, and Barnstable

C.J.

(2003b). Therapeutic prospects for PEDF: More than a promising angiogenesis inhibitor. Trends Mol. Med., 9, 244–250.

33.

Verma

I.M.

, and Somia

(1997). Gene therapy—promises, problems and prospects. Nature, 389, 239–242.

34.

Weis

S.M.

, and Cheresh

D.A.

(2011). Tumor angiogenesis: Molecular pathways and therapeutic targets. Nat. Med., 17, 1359–1370.

35.

Wright

J.F.

(2011). New adeno-associated virus strategies to support momentum in the clinic. Hum. Gene Ther., 22, 519–521.

36.

Q.J.

, Gong

C.Y.

, Luo

S.T.

, et al. (2012). AAV-mediated human PEDF inhibits tumor growth and metastasis in murine colorectal peritoneal carcinomatosis model. BMC Cancer, 12, 129.

37.

Xiao

, Li

, and Samulski

R.J.

(1996). Efficient long-term gene transfer into muscle tissue of immunocompetent mice by adeno-associated virus vector. J. Virol., 70, 8098–8108.

Supplementary Material

Please find the following supplemental material available below.

For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.

For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.

0.00 MB

0.31 MB

0.17 MB

0.05 MB

0.12 MB

0.14 MB