CAR T Cell Immortalization via hTERT and SV40LT
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
CAR T cell therapy faces challenges in maintaining long-term presence of CAR T cells, which is crucial for effective tumor treatment and preventing recurrence, as some patients struggle to generate enough T cells post-chemotherapy, risking their lives and potentially leading to tumor recurrence.
Innovation Solution
Integration of nucleic acid sequences encoding hTERT and SV40LT into CAR T cells, along with a suicide gene system, to enhance cell growth and longevity, and using an inducible expression system to regulate gene expression, combined with a regulatory compound that binds the CAR's extracellular domain to stimulate T cells and promote proliferation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If chemotherapy is administered to wipe out existing immune cells before CAR T cell therapy, then tumor burden is reduced and CAR T cell therapy can be initiated, but T cell counts drop significantly and may not recover sufficiently for continuous therapy
Solution Approach 1:
The patent introduces hTERT and SV40LT genes into T cells before chemotherapy to pre-establish immortalization capability. This preliminary genetic modification ensures that even after chemotherapy wipes out most T cells, the remaining or regenerated T cells possess the ability to proliferate indefinitely, thus maintaining sufficient T cell counts for continuous CAR T cell therapy.
Solution Approach 2:
The patent changes the biological parameter of T cell lifespan by introducing hTERT (telomerase reverse transcriptase) and SV40LT (simian virus 40 large T antigen) genes. These genetic modifications alter the fundamental aging parameter of T cells, transforming them from finite-lived cells to immortal cells that can persist and proliferate long-term in the patient's body, ensuring continuous anti-tumor activity.
2Reliability
If CAR T cells are infused to treat tumors, then tumor burden is reduced, but long-term maintenance of CAR T cells in patient bodies is insufficient leading to tumor recurrence
Solution Approach 1:
The patent fundamentally changes the lifespan parameter of CAR T cells by introducing hTERT and SV40LT genes. This genetic modification transforms CAR T cells from transient populations that typically decay over months to immortal populations that can persist indefinitely in the patient's body, thereby maintaining long-term tumor control and preventing recurrence.
Solution Approach 2:
The patent ensures continuous anti-tumor action by creating immortal CAR T cells through hTERT and SV40LT gene introduction. These modified cells can proliferate and persist continuously in the patient's body without requiring repeated infusions, maintaining constant surveillance and attack against tumor cells throughout the patient's life.
3Duration of action of moving object
If hTERT and SV40LT genes are introduced to enhance CAR T cell longevity, then cell growth and persistence are improved, but risk of uncontrolled proliferation and safety concerns increase
Solution Approach 1:
The patent employs a feedback control mechanism where CAR antigen recognition provides a self-regulating signal for hTERT and SV40LT expression. When CAR T cells encounter their target antigen, this triggers controlled expression of the immortalization genes, allowing proliferation only when needed for tumor control. When antigen is absent, gene expression is downregulated, preventing uncontrolled proliferation and ensuring safety.
Solution Approach 2:
The patent creates a dynamic system where the expression of hTERT and SV40LT genes is not constitutive but rather regulated in response to antigen presence. This dynamic regulation allows the system to adapt between states of controlled proliferation (when tumors are present) and quiescence (when tumors are controlled), balancing longevity benefits with safety concerns.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach results in a 1.5 to 3-fold increase in CAR T cell growth, maintaining cell density, and ensuring long-term in vitro survival, thereby enhancing the efficacy and longevity of CAR T cells for sustained tumor treatment.
Implementation Method 1
integration of the nucleic acid sequence encoding hTERT, a nucleic acid encoding SV40LT, or a combination thereof includes genomic integration of the nucleic acid sequence encoding hTERT, the nucleic acid encoding SV40LT, or a combination thereof
Implementation Method 2
constitutive expression of hTERT, SV40LT, or a combination thereof
Implementation Method 3
expression of hTERT, SV40LT, or a combination thereof, is regulated by an inducible expression system
Implementation Method 4
introducing a nucleic acid sequence encoding a suicide gene into the cell and culturing the CAR cell comprising the suicide gene and the nucleic acid encoding CAR with a nucleoside analogue in a manner permitting expression of the suicide gene to render the nucleoside analogue cytotoxic
Implementation Method 5
culturing the cells in the presence of an agent that the extracellular domain of the CAR recognizes
Data Source
Figure 1~2
Figure 3~4
Figure 5
AI summary
This invention provides a method for preparing CAR-T cells, comprising: providing cells comprising a chimeric antigen receptor; and culturing the cells in the presence of an agent that an extracellular domain of the CAR binds to obtain CAR cells, as well as the production thereof.