System biology analysis to develop diagnostic biomarkers, monitoring pathological indexes, and novel therapeutic approaches for immune targeting based on maggot bioactive compounds and polyphenolic cocktails in mice with gastric cancer.

Early diagnosis and prognosis are prerequisites for mitigating mortality in gastric cancer (GaCa). Identifying some causative or sensitive elements (coding RNA (cRNA)-non-cRNAs (ncRNAs)) can be very helpful in the early diagnosis of GaCa. Notably, despite significant development in the GaCa treatment, the outcome of patients does not remain satisfactory due to limitations such as multi-drug resistance and tumor relapse. Therefore, more attention has been drawn to complementary therapies and the use of supplements. In this regard, Polyphenol natural compounds (PNC) and maggot larvae (MaLa) alone or in combination were administered along with chemotherapy (paclitaxel) to N-methyl-N-nitrosourea (MNU)- induced murine tumor model. In addition, in order to identify potential diagnostic or prognostic biomarkers, transcriptomics analysis was performed through a bioinformatics approach. Then transcription profile of ncRNAs with their target hub genes was assessed through qPCR Real-Time, Western blot, and ELISA. According to the bioinformatics results, 17 hub genes (e.g., IL-6, CXCL8, MKI67, IL-2, IL-4, IL-10, IL-1ß, SPP1, LOX, COL1A1, and IFN-γ) were explored that contribute towards inflammation and oxidative stress and ultimately GaCa development. Upstream of the mentioned hub genes, regulatory factors (lncRNA XIST and NEAT1) were also identified and introduced as prognosis and diagnosis biomarkers for GaCa. Our results showed that PNC alone and in combination with MaLa was able to reduce the size and number of tumors, which is related to the reduction of genes expression levels (including IL-6, CXCL8, MKI67, IL-2, IL-4, IL-10, IL-1ß, SPP1, LOX, COL1A1, IFN-γ, NEAT1, and XIST). In conclusion, PNC and MaLa have the potential to be considered as complementary and improving chemotherapy due to their effective compounds. Also, the introduced hub gene and lncRNA in addition to diagnostic and prognostic biomarkers can be used as druggable proteins for novel therapeutic targeting of GaCa.

Production of a biosimilar version of aflibercept to improve VEGF blocker cytotoxicity on endothelial cells.

This project aimed to produce a biosimilar version of aflibercept (AFL) and evaluate the effect of the co-treatment of AFL with other vascular endothelial growth factor (VEGF) blocker drugs. For this purpose, the optimized gene was inserted into the pCHO1.0 plasmid and transfected into the CHO-S cell line. The final concentration of biosimilar-AFL for the selected clone was 782 mg/L. Results revealed that the inhibition potential of the biosimilar-AFL on HUVEC cells was significant at 10 and 100 nM concentrations and in a dose-dependent manner. Furthermore, co-treatment of biosimilar-AFL with Everolimus (EVR), Lenvatinib (LEN), and Sorafenib (SOR) could reduce HUVEC cell viability/proliferation, more than when used alone. When LEN and SOR were co-treated with biosimilar-AFL, their cytotoxicity increased 10-fold. The most and least efficient combination was seen when biosimilar-AFL combined with LEN and EVR, respectively. Finally, biosimilar-AFL may improve the efficiency of LEN, EVR, and SOR in reducing the VEGF effect on endothelial cells.

Triggering of lymphocytes by CD28, 4-1BB, and PD-1 checkpoints to enhance the immune response capacities.

Tumor infiltrating lymphocytes (TILs) usually become exhausted and dysfunctional owing to chronic contact with tumor cells and overexpression of multiple inhibitor receptors. Activation of TILs by targeting the inhibitory and stimulatory checkpoints has emerged as one of the most promising immunotherapy prospectively. We investigated whether triggering of CD28, 4-1BB, and PD-1 checkpoints simultaneously or alone could enhance the immune response capacity of lymphocytes. In this regard, anti-PD-1, CD80-Fc, and 4-1BBL-Fc proteins were designed and produced in CHO-K1 cells as an expression host. Following confirmation of the Fc fusion proteins' ability to bind to native targets expressed on engineered CHO-K1 cells (CHO-K1/hPD-1, CHO-K1/hCD28, CHO-K1/hCTLA4, and CHO-K1/h4-1BB), the effects of each protein, on its own and in various combinations, were assessed in vitro on T cell proliferation, cytotoxicity, and cytokines secretion using the Mixed lymphocyte reaction (MLR) assay, 7-AAD/CFSE cell-mediated cytotoxicity assay, and a LEGENDplex™ Human Th Cytokine Panel, respectively. MLR results demonstrated that T cell proliferation in the presence of the combinations of anti-PD-1/CD80-Fc, CD80-Fc/4-1BBL-Fc, and anti-PD-1/CD80-Fc/4-1BBL-Fc proteins was significantly higher than in the untreated condition (1.83-, 1.91-, and 2.02-fold respectively). Furthermore, anti-PD-1 (17%), 4-1BBL-Fc (19.2%), anti-PD-1/CD80-Fc (18.6%), anti-PD-1/4-1BBL-Fc (21%), CD80-Fc/4-1BBL-Fc (18.5%), and anti-PD-1/CD80-Fc/4-1BBL-Fc (17.3%) significantly enhanced cytotoxicity activity compared to untreated condition (7.8%). However, concerning the cytokine production, CD80-Fc and 4-1BBL-Fc alone or in combination significantly increased the secretion of IFN-γ, TNF-α, and IL-2 compared with the untreated conditions. In conclusion, this research establishes that the various combinations of produced anti-PD-1, CD80-Fc, and 4-1BBL-Fc proteins can noticeably induce the immune response in vitro. Each of these combinations may be effective in killing or destroying cancer cells depending on the type and stage of cancer.

Caspase-3: Structure, function, and biotechnological aspects.

Caspase-3, a cysteine-aspartic acid protease, has recently attracted much attention because of its incredible roles in tissue differentiation, regeneration, and neural development. This enzyme is a key zymogen in cell apoptosis and is not activated until it is cleaved by initiator caspases during apoptotic flux. Since caspase-3 has represented valuable capabilities in the field of medical research, biotechnological aspects of this enzyme, including the production of recombinant type, protein engineering, and designing delivery systems, have been considered as emerging therapeutic strategies in treating the apoptosis-related disorders. To date, several advances have been made in the therapeutic use of caspase-3 in the management of some diseases such as cancers, heart failure, and neurodegenerative disorders. In the current review, we intend to discuss the caspase-3's structure, functions, therapeutic applications, as well as its molecular cloning, protein engineering, and relevant delivery systems.

Tatibody, a recombinant antibody with higher internalization potency.

Using antibody drug conjugates (ADC) which can exclusively bind to their target cells and upon internalization release their toxic agent, is one of the most effective methods for killing tumor cells. Therefore, increasing the internalization rate is an important factor for tumor treatment in this case. The aim of the present study was to develop a new variant of pertuzumab (an anti-ErbB2 humanized antibody) with higher internalization rate that can be a good candidate for the production of ADC. To this end, the Human Immunodeficiency Virus TAT Protein Transduction Domain (TAT-PTD) was replaced into the structure of the pertuzumab. At first, the best site in antibody heavy chain constant region for the replacement of TAT-PTD was predicted through computational methods. Then, the resulting recombinant antibody, of which TAT-PTD was located at amino acid position 130-140 and named Tatibody, was produced in CHO-S cell line. Finally, its physicochemical properties and biological activities were evaluated and compared with pertuzumab. Results showed that the binding ability of Tatibody to the ErbB2 receptor is similar to that of pertuzumab, but its internalization potency is 3.6 fold higher and can be used as a good candidate for ADC construction.

Gold nanoparticles application in liver cancer.

Studies have shown that hepatocellular carcinoma (HCC) (the most important type of liver cancer) is the fifth most common cancer, and the third cause of mortality, globally. Although major progress has been made in the treatment and diagnosis of this disease, its eradication remains limited. Consequently, discovering new diagnosis and treatment methods is important. Cancer nanotechnology is an emerging field in medicine with the aim to accomplish advances in both cancer diagnosis and treatment. Gold nanoparticles (GNPs/ AuNPs) have attracted much attention, owing to their biocompatibility (bio-inertness, and low cytotoxicity), ability to chemically modify their surface by attaching multiple types of ligands, and their superior optical properties. This review will focus on the current applications of AuNPs in different aspects, such as imaging, drug and gene delivery, radiotherapy, and photothermal therapy of liver cancer treatment.

Phytoremediation of Ni-contaminated soil by Salicornia iranica.

Although nickel (Ni) is useful and is used in various industries, it is one of the most usual and important sources of heavy metals pollutants in the world. In this study, Salicornia iranica was used in order to phytoremediate Ni-contaminated soil. Possible mechanisms of plant tolerance to Ni pollution and its detoxification were studied through using expression analysis of glutathione-S-transferase (GST) and measurement of involved key physiological components. The concentration of the chlorophylls a, b, total chlorophyll, and carotenoids were significantly decreased in 500 mg/kg Ni at 3, 24, 48 h, and 90 days after the treatment. Free proline significantly increased in the tissues. The absorption and concentration of Ni increased in tissues, so that Ni concentration at 50, 250, and 500 mg Ni/kg soil significantly increased to 2.5, 3.5, and 4.5 fold compared with the lowest Ni level respectively. In addition, the GST expression was significantly increased both in the 50 and 500 mg/kg Ni treatment. The highest concentration of Ni affected plant growth parameters such as the root and shoot lengths. Therefore, S. iranica is able to accumulate Ni and it can be used as an environmental biotechnological study for phytoremediation of Ni-polluted soils. Abbreviations: ABA: abscisic acid; ABRE: ABA-responsive element; As+3: arsenic; Cd2+: cadmium; ef1: elongation factor; FW: fresh weight; GSH: glutathione; GST: glutathione-S-transferase; GSTU: tau class GST; Hcl: hydrochloric acid; Hg2+: mercury; HgCl2: mercury(II) chloride; MYB: myeloblastosis viral oncogene homolog; Ni+2: nickel; Pb: lead; SiGSTU: Salicornia iranica GSTU; ZnSO4: zinc sulfate.

Phytoremediation of petroleum-contaminated soil by Salicornia: from PSY activity to physiological and morphological communications.

Petroleum is one of the critical environmental pollutants. Salicornia can grow in petroleum-contaminated soil. Therefore, the potential of two Iranian Salicornia species, S. persica Akhani and S. iranica Akhani, for phytoremediation of soils contaminated with 0.2% or 2% petroleum was evaluated over short (1 and 10 h) and long (100 days) periods of time. In addition, some key factors including the expression analysis of phytoene synthase, physiological and morphological factors were studied. Both species reduced the petroleum in 0.2% and 2% petroleum-contaminated soils to 40% and 60% of the initial amount, respectively. The expression of PSY increased twice more than the control 10 h after 0.2% petroleum stress and the carotenoid content increased twice more than the control. Chlorophyll a and total chlorophyll decreased three times less than the control in both contamination levels, while chlorophyll b decreased three times less than the control only in 2% contamination. The proline content peaked 10 h after 2% stress as it was 10 times more than the control. Promoter analysis of PSY showed the existence of responsive cis-acting elements to abscisic acid suggesting the key role of this gene in abiotic stresses.