Expression of plant-produced anti-PD-L1 antibody with anoikis sensitizing activity in human lung cancer cells via., suppression on epithelial-mesenchymal transition

Abstract

Immune checkpoint antibodies in cancer treatment are receptor-ligand pairs that modulate cancer immunity. PD-1/PD-L1 pathway has emerged as one of the major targets in cancer immunotherapy. Atezolizumab, the first anti-PD-L1 antibody approved for the treatment of metastatic urothelial, non-small cell lung, small cell lung and triple-negative breast cancers, is produced in Chinese Hamster Ovary (CHO) cells with several limitations i.e., high-production costs, low-capacity yields, and contamination risks. Due to the rapid scalability and low production costs, the transient expression in Nicotiana benthamiana leaves was investigated by co-infiltration of Agrobacterium tumefaciens GV3101 cultures harboring the nucleic acid sequences encoding for Atezolizumab heavy chain and light chain in this study. The transient expression of Atezolizumab in transformed N. benthamiana accumulated up to 86.76 μg/g fresh leaf weight after 6 days of agroinfiltration (OD 600 nm: 0.4) with 1:1 ratio of heavy chain to light chain. The structural and functional characteristics of plant-produced Atezolizumab was compared with commercially available Tecentriq® from CHO cells with similar binding efficacies to PD-L1 receptor. The direct anti-cancer effect of plant-produced anti-PD-L1 was further performed in human lung metastatic cancer cells H460 cultured under detachment condition, demonstrating the activity of anti-PD-L1-antibody on sensitizing anoikis as well as the suppression on anti-apoptosis proteins (Bcl-2 and Mcl-1) and modulation of epithelial to mesenchymal regulating proteins (E-cadherin, N-cadherin, Snail and Slug). In conclusion, this study manifests plants as an alternative cost-effective platform for the production of functional monoclonal antibodies for use in cancer therapy.

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

“…Our findings demonstrate that a recombinant bsAb can be produced transiently in plants within 4 days, with up to 81.63 ± 17.56 µg of DVD-AT2C obtained per g of fresh leaf mass. This data is consistent with our prior studies [23,24] for producing plant-based antibodies in less than a week, which is faster than other expression platforms [25] and transgenic plants [26].…”

“…Plant-produced bsAb DVD-AT2C was further assessed by an SEC method using a 2.5-µM particle size SEC BEH 200 column (4.6 × 300 mm; Waters) mounted on a UHPLC system (Waters), as previously described [21,23]. A bsAb protein sample was injected onto the purification system and eluted with PBS buffer (pH 7.4).…”

Anoikis‐related genes as potential prognostic biomarkers in gastric cancer: A multilevel integrative analysis and predictive therapeutic value

“…Other investigations discovered that the overexpressed anoikis gene CEACAM6 was not only oncogenic, but its monoclonal antibodies and fragments acted as immune checkpoint inhibitors in haematological malignancies [37]. And the anti-PD-L1 antibody had anoikis sensitising activity to suppress EMT in human lung cancer cells [38]. Due to the anti-apoptotic phenomenon and significant immune gene expression differences, the mechanism of cancer metastasis away from the anchorage-dependent cell cycle was probably linked to the escape of TIICs and immune checkpoints.…”

A novel anoikis-related gene signature to predict the prognosis, immune infiltration, and therapeutic outcome of lung adenocarcinoma

“…Cancer cells could overcome anoikis which would contribute to tumor progression, metastasis, and resistance of therapy (21). Previous studies (13,22) have demonstrated that anoikis plays great roles in the development of lung cancer. Cellular retinoic acid binding protein 2 (CRABP2) was shown to promote integrin β1/FAK/ERK signaling and inhibit anoikis; overexpressed CRABP2 was found to increase lymph node and liver metastasis through in vivo tail vein injection of lung cancer cell models (23).…”