Generation and characterization of novel anti-DR4 and anti-DR5 antibodies developed by genetic immunization

Abstract

Development of therapeutic antibodies in oncology has attracted much interest in the past decades. More than 30 of them have been approved and are being used to treat patients suffering from cancer. Despite encouraging results, and albeit most clinical trials aiming at evaluating monoclonal antibodies directed against TRAIL agonist receptors have been discontinued, DR4 or DR5 remain interesting targets, since these receptors are overexpressed by tumour cells and are able to trigger their death. In an effort to develop novel and specific anti-DR4 and anti-DR5 antibodies with improved properties, we used genetic immunization to express native proteins in vivo. Injection of DR4 and DR5 cDNA into the tail veins of mice elicited significant humoral anti-DR4 and anti-DR5 responses and fusions of the corresponding spleens resulted in numerous hybridomas secreting antibodies that could specifically recognize DR4 or DR5 in their native forms. All antibodies bound specifically to their targets with a very high affinity, from picomolar to nanomolar range. Among the 21 anti-DR4 and anti-DR5 monoclonal antibodies that we have produced and purified, two displayed proapoptotic properties alone, five induced apoptosis after cross-linking, four were found to potentiate TRAIL-induced apoptosis and three displayed antiapoptotic potential. The most potent anti-DR4 antibody, C#16, was assessed in vivo and was found, alone, to inhibit tumour growth in animal models. This is the first demonstration that DNA-based immunization method can be used to generate novel monoclonal antibodies targeting receptors of the TNF superfamily that may constitute new therapeutic agents.

Tumor Necrosis Factor Family Member Profile Predicts Prognosis and Adjuvant Chemotherapy Benefit for Patients With Small-Cell Lung Cancer

“…TNFRSF10B (DR5) is primarily localized on the plasma membrane, where it functions as an apoptotic receptor to induce cell apoptosis. A DR5-targeting antibody has been developed and tested in clinical trials as a therapeutic intervention for cancer patients ( 50 , 51 ). TNFSF14 (LIGHT, HVEML, CD258), a secreted protein, is a highly effective stimulator of antitumor immune responses, which also influences the plasticity of the TIME ( 52 ).…”

First-in-human DR5 PET reveals insufficient DR5 expression in patients with gastrointestinal cancer

“…In the current study, over 50.0% (10/18) of patients had no DR5 expression, and only 16.7% (3/18) of Open access patients had DR5 overexpression in tumors, which indicated that the rate of DR5 expression was not as high as previously reported. 26 This finding may also be the reason why only a subset of patients in the previous clinical trials showed antitumor effects, and no trials could achieve the endpoint of improving the efficacy in unselected patients. 27 It might be because of the inaccurate detection of DR5 by IHC in previous studies, resulting in the overestimation of the DR5 expression rate in tumors.…”

CD13 inhibition augments DR4-induced tumor cell death in a p-ERK1/2-independent manner

“…To date, dulanermin (AMG 951), a recombinant soluble TRAIL, is in an ongoing phase III trial, and phase I and phase II trials have been completed 13,14 . In addition, 7 tests have been performed on DR agonistic monoclonal antibodies, including mapatumumab (HGS-ETR1), the only DR4 targeting antibody assessed in phase I/II trials 15 . Although these treatments have exhibited efficacy in triggering apoptosis for sensitizing tumor cells to radiotherapy, targeted therapy and chemotherapy, the outcomes have been disappointing, owing to the development of intrinsic and/or acquired resistance in many tumor cell lines; this drawback remains a crucial challenge for researchers 16,17 .…”