Immunoglobulin Fc Domain Fusion to TRAIL Significantly Prolongs Its Plasma Half-Life and Enhances Its Antitumor Activity

Abstract

TRAIL (Apo2L) is a potent inducer of cell death. Interest in TRAIL has increased, following the observation that TRAIL can selectively kill a wide variety of human cancer cells without killing normal cells both in vitro and when grown as xenografts. Therefore, TRAIL has been proposed as a promising anticancer agent and currently is being tested in clinical trials. However, recombinant TRAIL has a very short plasma half-life, which limits its therapeutic potential. To overcome this limitation, we investigated the ability of the human IgG1 fragment crystallizable region (Fc) to enhance TRAIL stability. In this report, we show that Fc-TRAIL chimeric protein displays higher specific activity in vitro and a significantly longer half-life in mice than recombinant human TRAIL (rh-TRAIL). No short-term toxicity, especially liver toxicity, was observed. More importantly, Fc-TRAIL was much more effective in inhibiting tumor growth in a xenograft tumor model compared with rh-TRAIL. Our data suggest that fusion of Fc to TRAIL is able to improve the bioavailability and activity of TRAIL both in vitro and in vivo, and Fc-TRAIL may be explored for future clinical applications in cancer treatment and prevention. Mol Cancer Ther; 13(3); 643-50. Ó2014 AACR.

Recombinant human CD19L-sTRAIL effectively targets B cell precursor acute lymphoblastic leukemia

“…reported to have a modestly prolonged plasma half-life of 100 minutes (43). Despite its larger size, which should limit its renal clearance, that Fc fusion protein was likely cleared by the Fc receptorpositive elements of the reticuloendothelial system.…”

Preclinical study of rAAV2-sTRAIL: pharmaceutical efficacy, biodistribution and safety in animals

“…Briefly, BALB/c (nu/nu) nude mice were subcutaneously injected at the right axillary space with 0.1 ml of cell suspension containing 2 × 10 6 tumor cells. When tumors were grown up to about 300 mm 3 , the animals were killed. The tumors were excised and cut into small pieces with about 2 mm diameter, and then subcutaneously inoculated at the right axillary space of another group of mice.…”

“…The tumors were excised and cut into small pieces with about 2 mm diameter, and then subcutaneously inoculated at the right axillary space of another group of mice. When the size of xenografts reached about 100 mm 3 , a single dose of rAAV2-TRAIL virus at 1 × 10 9 , 1×10 10 and…”

“…It was shown that neither Ad5-null nor AAV2-null virus possessed an influence on the tested tumor cell growth (data not shown), indicating that the cytotoxicity mediated by rAAV2-sTRAIL virus infection was caused by AAV-mediated sTRAIL expression, not by the AAV2-null or Ad5-null helper virus. Antitumor efficacy in mouse xenografts of human cancers To explore the antitumor efficacy of rAAV2-sTRAIL virus infection, when the size of the xenograft of HCT-116, NCI-H460 and BEL-7402 cancer cells reached about 100 mm 3 , a single dose of rAAV2-sTRAIL virus (1 × 10 9 , 1 × 10 10 and 1 × 10 11 vg) was injected (s.c.), respectively. Then tumor growth was monitored for 19-24 days depending on the mouse life conditions.…”

“…A couple of recombinant soluble TRAIL has been tested in phase I and/or II clinical trials; however, the very short half-life (3-5 min in rodents and 23-31 min in non-human primates) limited its therapeutic potentials. 3 TRAIL derivants such as iz-TRAIL, 4 TNC-TRAIL, 5 HSA-TRAIL, 6 PEG-TRAIL 7,8 , etc., prolong the half-life and enhance stability. 9 However, the outcomes of monodrug of rTRAIL derivants in the clinical trials for cancer therapy still were not satisfied.…”