The biological activity of bispecific trastuzumab/pertuzumab plant biosimilars may be drastically boosted by disulfiram increasing formaldehyde accumulation in cancer cells

Abstract

Studies of breast cancer therapy have examined the improvement of bispecific trastuzumab/pertuzumab antibodies interacting simultaneously with two different epitopes of the human epidermal growth factor receptor 2 (HER2). Here, we describe the creation and production of plant-made bispecific antibodies based on trastuzumab and pertuzumab plant biosimilars (bi-TPB-PPB). Using surface plasmon resonance analysis of bi-TPB-PPB antibodies binding with the HER2 extracellular domain, we showed that the obtained Kd values were within the limits accepted for modified trastuzumab and pertuzumab. Despite the ability of bi-TPB-PPB antibodies to bind to Fcγ receptor IIIa and HER2 oncoprotein on the cell surface, a proliferation inhibition assay did not reveal any effect until α1,3-fucose and β1,2-xylose in the Asn297-linked glycan were removed. Another approach to activating bi-TPB-PPB may be associated with the use of disulfiram (DSF) a known aldehyde dehydrogenase 2 (ALDH2) inhibitor. We found that disulfiram is capable of killing breast cancer cells with simultaneous formaldehyde accumulation. Furthermore, we investigated the capacity of DSF to act as an adjuvant for bi-TPB-PPB antibodies. Although the content of ALDH2 mRNA was decreased after BT-474 cell treatment with antibodies, we only observed cell proliferation inhibiting activity of bi-TPB-PPB in the presence of disulfiram. We concluded that disulfiram can serve as a booster and adjuvant for anticancer immunotherapy.

Development of Plant-Derived Bispecific Monoclonal Antibody Targeting PD-L1 and CTLA-4 against Mouse Colorectal Cancer

“…Plant transformation has also been uti-lized to express bifunctional antibodies specific for CHIKV and DENV, and the plant-made bsAb demonstrated potent neutralization of both viruses [31]. Further, transient expression of bispecific trastuzumab and pertuzumab plant biosimilars (bi-TPB-PPB) was also reported in N. benthamiana, and bi-TPB-PPB and disulfiram treatment successfully inhibited breast cancer cell proliferation with even higher efficiency than trastuzumab or pertuzumab plant biosimilars [13]. Thus, this research additionally establishes the robustness of plant expression systems for the production of high-value complex immune-oncology antibody therapeutics.…”

“…Many reports have shown that plants can be an efficient system for expressing various types of antibodies, including secretory immunoglobulins (IgG and IgA) [27,28] and bsAbs [13,29]. For example, secretory IgA (sIgA) variants of ustekinumab and 2G12 antibodies against inflammatory bowel disease and HIV have been made in planta [28,30].…”

“…On the other hand, plants represent an alternative platform for producing antibodies due to their ability to synthesize proteins and perform posttranslational modifications somewhat similar to animal cells [13,14]. In addition, plant systems have several advantages when used as a factory for pharmaceutical protein production, such as low risk of contamination with adventitious agents, simplicity, and low production cost [13]. In fact, the cost of growing the plants commonly used for protein manufacturing is far cheaper than the cost of cultivating other expression systems [15][16][17].…”

A Physiologically Based Pharmacokinetic Framework for Quantifying Antibody Distribution Gradients from Tumors to Tumor-Draining Lymph Nodes

“… R01: baseline target concentration in PT [ 29 ]; R02: baseline target concentration in TDLN [ 29 ]; K d : antibody affinity to target [ 30 , 31 , 32 ]; K d,FcRn : antibody affinity to FcRn [ 33 ]; dln: shape factor [ 28 , 34 ]; CL p : antibody clearance from plasma [ 35 , 36 , 37 , 38 ]. Antibody/tumor specific parameters that were shared by all organ systems include: k deg : target degradation rate, 0.01 1/h; k int : antibody-target complex internalization rate, 0.01 1/h.…”

Teaching new tricks to old dogs: A review of drug repositioning of disulfiram for cancer nanomedicine

“…Besides, the tumor cell (BT‐474) proliferation was inhibited by trastuzumab and pertuzumab plant biosimilars in combination with DSF that served as a booster and adjuvant for the therapeutic antibodies. [ 49 ] Therefore, DSF‐based FROUNT targeting is a promising approach for cancer immunotherapy via regulating TAMs.…”