Selection and Characterization of a Novel Agonistic Human Recombinant Anti-Trail-R2 Minibody with Antileukemic Activity

Abstract

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising natural anticancer therapeutic agent because through its death receptors, TRAIL-R1 and TRAIL-R2, it induces apoptosis in many transformed tumor cells, but not in the majority of normal cells. Hence, agonistic compounds directed against TRAIL death receptors have the potential of being excellent cancer therapeutic agents, with minimal cytotoxicity in normal tissues. Here, we report the selection and characterization of a new single-chain fragment variable (scFv) to TRAIL-R2 receptor isolated from a human phage-display library, produced as minibody (MB), and characterized for the in vitro anti-leukemic tumoricidal activity. The anti-TRAIL-R2 MB2.23 efficiently and specifically bound to membrane-associated TRAIL-R2 on different leukemic cell lines and could act as a direct agonist in vitro, initiating apoptotic signaling as well as complement-dependent cytotoxicity and antibody-dependent cell cytotoxicity, providing a rationale for further investigations of MB2.23 in anticancer therapy.

Nanoparticles Engineered with Rituximab and Loaded with Nutlin-3 Show Promising Therapeutic Activity in B-Leukemic Xenografts

“…For complement-dependent cytotoxicity (CDC) assay, experiments were carried out as previously described (28,29) with some modifications. After cell exposure to the different NP preparations and after the addition of pooled normal AB HS (25%), cultures were incubated at 37 C for 24 and 48 hours before determination of the percentage of induced dead cells.…”

In vivo anti-lymphoma activity of an agonistic human recombinant anti-TRAIL-R2 minibody

“…P values were calculated by long-rank test. Differences were considered significant when P value was <0.05. expressed in CHO cells and purified by affinity column chromatography as previously described [5]. The in vivo evaluation of the potential anti-tumor activity of MB2.23 was investigated by using human disseminated lymphoma bearing SCID mouse xenografts.…”

“…After production in CHO cells and purification from culture supernatants, the previously characterized MB2.23 [5] was diluted in sterile PBS and dosed at 10 mg/kg and for the intraperitoneal (i.p.) injections.…”

“…In a recent study, we have described a human anti-TRAIL-R2 minibody, named MB2.23, which specifically recognizes and binds TRAIL-R2 without exhibiting crossreactivity with TRAIL-R1 [5]. In the present study, we have investigated the MB2.23 minibody for its tumoricidal properties in vivo, in a xenograft model of lymphoma.…”

Prediction and extraction of microRNA2target interactions associated with leukemia

“…Nuclear accumulation of STAT5A as a result of activation by FLT3-ITD is an oncogene found in acute myeloid leukemia (Chatain et al, 2012). TNFRSF10B-induced apoptosis in chronic lymphocytic leukemia cells is one of the most promising candidates for cancer therapeutics (Natoni et al, 2007;Secchiero et al, 2009). MAPK1 is involved in a wide variety of cellular processes, such as proliferation, differentiation, transcription regulation and development, correlated with acute myelogenous leukemia (Vey et al, 2004).…”