CAR T-Cell Control via Knottin Proteins and RNA Destabilizing Elements
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current CAR T-cell therapies face challenges with dangerous side effects such as cytokine release syndrome, tumor lysis syndrome, B-cell aplasia, and on-tumor, off-target toxicities due to the inability of existing kill switches and transient CAR expression strategies to effectively control CAR T-cell activity.
Innovation Solution
The use of knottin proteins as antigen recognition components in CARs, combined with RNA Destabilizing Elements (RDEs) to regulate transgene expression, allowing for precise control of CAR T-cell activity through metabolic and ligand-induced mechanisms, and combination therapies with other treatments like chemotherapy and antibody-drug conjugates.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Object-affected harmful factors
If kill switch technology is used to control CAR T-cell activity, then toxicity is reduced, but the long-term surveillance benefit is sacrificed
Solution Approach 1:
The patent applies dynamics by making CAR expression controllable and adjustable rather than fixed. Through inducible promoters and regulatory elements, the system can dynamically adjust CAR expression levels in response to therapeutic needs, allowing simultaneous achievement of toxicity control and long-term surveillance capability.
Solution Approach 2:
The patent changes the parameter of CAR expression control from binary (on/off) to graded and temporally controlled. By using promoters with different induction characteristics and regulatory elements that modulate expression levels, the system achieves precise control over CAR activity to balance toxicity prevention with surveillance function.
2Object-affected harmful factors
If transient CAR expression is used to control toxicity, then CAR exposure is limited, but the surveillance benefit is sacrificed
Solution Approach 1:
The patent implements periodic action through temporally controlled CAR expression. CARs are expressed in controlled pulses or cycles rather than continuously, allowing the system to provide surveillance function during appropriate time windows while limiting overall CAR exposure to prevent toxicity.
Solution Approach 2:
The system transitions from static CAR expression to dynamic, time-dependent expression patterns. Through inducible promoters and regulatory mechanisms, CAR expression can be activated and deactivated at specific times, enabling both toxicity control and sustained surveillance capability when needed.
3Duration of action of stationary object
If stable transgene insertion is used to ensure long-term CAR expression, then surveillance benefit is maintained, but error-prone retroviral copying occurs
Solution Approach 1:
The patent extracts the problematic element (retroviral copying mechanism) from the system and replaces it with alternative methods of transgene delivery and expression control. By using non-retroviral vectors or integrated expression systems that avoid error-prone copying, the system maintains long-term surveillance while improving insertion accuracy.
Solution Approach 2:
The patent uses precise copying mechanisms for transgene integration that minimize errors. Through improved viral vectors or non-viral delivery methods with higher fidelity, the system achieves accurate transgene insertion and maintenance over time, reducing mutational errors while preserving long-term CAR expression.
4Reliability
If knottin proteins are used as antigen recognition components, then binding affinity and stability are improved, but device complexity increases
Solution Approach 1:
The patent applies local quality by optimizing specific regions of the CAR construct. The knottin-based antigen recognition domain is designed with specific structural features (cysteine knot architecture) that provide enhanced binding affinity and stability, while other portions of the CAR remain streamlined to minimize overall complexity.
Solution Approach 2:
The patent uses composite material principles in protein design by combining the knottin scaffold structure with specific peptide loops and functional domains. This composite approach creates a modular CAR construct where each component serves a specific function, achieving high binding affinity while maintaining manageable complexity through standardized modular architecture.
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 reduces immunogenicity and toxicity, enhances the dynamic range of CAR expression, and allows for targeted and controlled delivery of therapeutic payloads to tumor sites, improving treatment efficacy while minimizing side effects.
Implementation Method 1
RNA Destabilizing Elements (RDEs) to regulate transgene expression
Implementation Method 2
Knottins can be engineered to recognize targets of interest as knottins have a core structure (scaffold) with peptide loops around the core structure which peptide loops can be engineered to produce different binding properties and specificities
Implementation Method 3
AP1903 (also known as rimiducid), a lipid-permeable tachrolimus analog that initiates homodimerization of the human protein FKBP12 (Fv)
Data Source
AI summary
Control Devices are disclosed including RNA destabilizing elements (RDE) combined with transgenes, including Chimeric Antigen Receptors (CARs) in eukaryotic cells. These RDEs can be used to optimize expression of transgenes, e.g., CARs, in the eukaryotic cells so that, for example, effector function is optimized. CARs and transgene payloads can also be engineered into eukaryotic cells so that the transgene payload is expressed and delivered at desired times from the eukaryotic cell. Such CAR T-cells with transgene payloads can be combined with the administration of other molecules, e.g., other therapeutics such as anticancer therapies.