Part

New parts we designed：

You can get detailed information on our Part collection page.

What our parts can do：

As we have mentioned in part collection, our basic part collection is mainly composed of two groups: the components of CAR and the components of our genetic circuit, namely, the kill-switch circuit. All parts will be available for future teams to help explore and construct new parts. Among these parts, we like BBa K4421003 (scfv library with 10 individual colonies) most, which provides novel ideas for whoever interested in CAR and tumor heterogeneity.

1) Scfv library with 10 individual colonies demonstrates the concept of antibody library. As we all know, developing and evolving tumors contain diverse and substantial antigens, which form an antigen library, rather than a single kind of antigen, embodying the characteristic of tumor heterogeneity. So the traditional ideas to depend on specific CARs are unreliable and inappropriate. In our project, we used an antibody library to combat with the antigen library extensively, addressing tumor heterogeneity successfully. Future teams or researchers who are devoted to tumor heterogeneity and CAR, can borrow from our part BBa_K4421003 and idea of antibody library.

2) Also, our kill switch part may help future teams to construct their own genetic circuits.

Our improvement of an existing part：

Based on BBa K3244013 and BBa K3755010, we added a new part BBa K4421031(improved NFAT response element), which was random mutanted from BBa_K3755010, but with a much higher response strength.

part contribution

When searching for project-related part information in the registry, we found in the part BBa K4040034 we can add new information about its usage.

Here shows what we added:

Usage supplementation: HER2, also known as ErbB2, is one of four members (HER1–4) of the epidermal growth factor receptor or HER family. All HER receptors share a similar structure: an extracellular ligand-binding domain, a short hydrophobic transmembrane region, and a cytoplasmic tyrosine kinase domain. Hetero- or homodimerization of HER receptors, induced by ligand binding or receptor overexpression, leads to the activation of the receptor kinase and to the subsequent phosphorylation of several tyrosine residues. In turn, these phosphorylated tyrosine residues, located within the carboxyl terminus of the receptors, recruit mediators and activate signaling pathways that result in the modification of the cell growth, differentiation, and survival.

When referencing as truncated receptor, it is usually means the p95HER2, which has kinase activity in the absence of the trastuzumab-binding extracellular domain led us to hypothesize that p95HER2–expressing tumors may be resistant to trastuzumab but sensitive to the inhibitory effects of lapatinib, a low–molecular-weight tyrosine kinase inhibitor (TKI) of HER2 that has activity in patients with HER2–expressing tumors that are resistant to trastuzumab.

However, this part includes the Truncated HER2 without the ICD domain, which can be used as a marker for engineered synthetic immune cells or other cells, such as CAR-T cells or liver cells. When used as a marker, the engineered cell can be easily cleared with anti-HER2 antibodies, such as Trastuzumab,or other HER2 targeting methods.

Model

①Bioinformatics was used to analyze the relationship between the number of CAR species on the likelihood of normal cell misadventure and the antigen escape rate, systematically demonstrating that a single cell cannot express too many CARs. For future teams to design CAR cells, the range of cellular loads is provided

② Modeling of CAR-NK92 signaling pathway: to address the structural differences between CAR and TCR, the biological behavior of CAR during antigen recognition is mathematically modeled on the basis of the kinetic proofreading model of TCR. It provides a mathematical strategy and a theoretical basis for the future team to predict the timing and efficiency of CAR activation gene circuits.

③Application of AP1903：based on drug dynamics and experimental data, a scheme is designed to artificially control the life cycle of CAR-NK92 cells with AP1903. It provides practical examples and new ideas for future teams to design human-cell interaction systems.

④Simulation of CAR antibody library coverage: A new mathematical idea for predicting the antigen coverage of antibody libraries is provided.

⑤ Metacellular automata simulation software and proliferation model：Based on CAR-NK92's own proliferation characteristics and interaction with tumor cells, an innovative visualization simulation experiment is conducted. For cell therapy of tumors, the program based on this simulation experiment can visualize and predict the characteristics of different therapeutic cells and their interaction with tumors. The simulation experiment and visualization program can provide prediction of the results of stem experiments for cell therapy wet experiments, which has great potential for extensive screening of cell therapy strategies and cost savings in cell therapy experiments.

Product

In this project, we connected the binding status of the receptor with the apoptosis signal of CAR-NK cells through the gene circuit, and controlled the dynamic proliferation process of CAR-NK cells with the dynamic change of tumor cell membrane antigen, so as to specifically enrich CAR-NK cells that can recognize and kill tumor cells. If the peripheral blood of the patient is extracted for analysis at this time, we can obtain the information of the tumor target. We can discover the potential tumor antigen.

The ideal process is as follows:

① We can extract the peripheral blood of the patient, in which the surviving NK cells can bind to tumor antigens of the patient.

② The NK cells in the peripheral blood will be isolated and purified by flow cytometry.

③We will extracted total mRNA of NK cells and obtain the cDNA by reverse transcription PCR. Then we will analyze the sequence information.

④Tumor target information can be used to develop monoclonal antibody drugs.