Gymnopoulos et al. 2020 aimed to investigate the effectiveness of a novel anticancer treatment called TR1801-ADC in patient-derived xenograft (PDX) models of solid tumors. In order to achieve this, several experiments were conducted based on various laboratory techniques and assays. In this context, a simplified explanation of these methods will be provided to enhance a better understanding of individuals not conversant with the medical and cancer treatment terms.
Patient-Derived Xenograft (PDX) Models:
Patient Derived Xenograft model, a tumor sample is obtained from patients with solid tumors and implanted into mice. The model helps create PDX models, which are mice with human tumor tissues growing in them (Gymnopoulos et al., 2020). These PDX models serve as a representative model of human (TR1801-ADC) on real human tumor tissues.
In this method, a human tumor cell is taken to the laboratory. The whole process involves providing the cells with an environment rich in nutrients and optimum temperature conditions to help the cancer cells grow and multiply.
Antibody-Drug Conjugate (ADC) Production
Another TR1801-ADC, or anticancer drug, was developed as a combination of an antibody and an anticancer drug. Antibodies are proteins produced by the body’s immune system that can specifically recognize and bind to certain molecules, such as cancer cells (Gymnopoulos et al., 2020). The potent anticancer drug is attached to the antibody, forming the Antibody-Drug Conjugate. This combination allows the ADC to target cancer cells specifically and deliver the drug directly to them.
In Vitro Assays
Another strategy used was the vitro assays to assess the efficacy and specificity of TR1801-ADC. One of the assays conducted was a cell viability assay, which measures the ability of TR1801-ADC to kill cancer cells while protecting normal cells (Gymnopoulos et al., 2020). An apoptosis assay was also conducted to determine whether TR1801-ADC induces programmed cell death in cancer cells. Apoptosis is the death of cells which is programmed by the body.
Additionally, Western blotting is a laboratory technique used to detect specific proteins in a sample. This technique analyzes the expression levels of the cMet protein in cancer cells before and after treatment with TR1801-ADC; cMet is a cell that promotes cancer development. The process involves separating proteins based on size using a gel and transferring them onto a special (PVDF) membrane.PVDF is a membrane used for identifying antigens of any disease causative agent and then probing the membrane with antibodies that can specifically recognize the cMet protein(Gymnopoulos et al., 2020). The process allows evaluation of TR1801-ADC’s impact on the presentation of the cMet protein in cancer cells.
In vivo experiments, the PDX models were used to evaluate the efficacy of TR1801-ADC in solid tumors. The PDX models were treated with TR1801-ADC, and the tumor growth was monitored over time. PDX models are tumors initially implanted in mice (Gymnopoulos et al., 2020). The tumor size is measured and assessed for other characteristics to determine the treatment’s effectiveness in inhibiting tumor growth.
Flow cytometry is a technique used to analyze different characteristics of cells. In this study, flow cytometry was used to evaluate the binding of TR1801-ADC to cancer cells (Gymnopoulos et al., 2020). It allowed the researchers to determine the percentage of cancer cells specifically targeted and bound by TR1801-ADC.
Pharmacokinetics studies how a drug is absorbed, distributed, and removed from the body. This study performed pharmacokinetic analysis to understand the behavior of the body’s anticancer therapy (TR1801-ADC) (Gymnopoulos et al., 2020). Blood samples are collected at different times after administering TR1801-ADC to the PDX models, and the concentration of the drug in the blood is tested or screened.
Immunohistochemistry is a technique for visualizing specific proteins or molecules in tissue samples. In this study, immunohistochemistry was performed to examine the presentations and location of the cMet protein in tumor tissues (Gymnopoulos et al., 2020). Tumor tissue sections were treated with specific antibodies, which produced a visible signal indicating the presence and distribution of cMet in the tissue.
In conclusion, this method provided a more comprehensive understanding of TR1801-ADC’s interaction with cancer cells, its distribution in the body, and the expression of the cMet protein in tumor tissues. These study objectives were to gain awareness of the potential of TR1801-ADC as a highly potent anticancer treatment for solid tumors. The study provides valuable information for further development and clinical evaluation of this antibody-drug conjugate in the fight against cancer.
Gymnopoulos, M., Betancourt, O., Blot, V., Fujita, R., Galvan, D., Lieuw, V., … & Coronella, J. (2020). TR1801‐ADC: a highly potent cMet antibody–drug conjugate with high activity in patient‐derived xenograft models of solid tumors. Molecular oncology, 14(1), 54–68. https://febs.onlinelibrary.wiley.com/doi/abs/10.1002/1878-0261.12600
Gymnopoulos, M., Betancourt, O., Blot, V., Fujita, R., Galvan, D., Lieuw, V., … & Coronella, J. (2020). TR1801‐ADC: a highly potent cMet antibody–drug conjugate with high activity in patient‐derived xenograft models of solid tumors. Molecular oncology, 14(1), 54–68.https://febs.onlinelibrary.wiley.com/doi/abs/10.1002/1878-0261.12600