Engineered B Cell Receptors With Targeted IgH Insertion for Natural Activation
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Solution Overview
Problem
Existing methods for genetic engineering of B cell receptors (BCRs) in B cells fail to achieve natural-like antigen-induced activation, affinity maturation, class switch recombination, and memory retention, often due to promiscuous integration and lack of native regulation, limiting the effectiveness of monoclonal antibody therapy.
Innovation Solution
A method involving targeted insertion of a nucleic acid cassette with a nucleic acid sequence coding for a variable domain of an immunoglobulin heavy chain and splice donor/acceptor sites into the IgH locus, enabling antigen-induced B cell activation and affinity maturation, using CRISPR/Cas9-mediated genome editing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If lentiviral vectors are used for genetic engineering of B cells, then antibody secretion is achieved, but promiscuous integration occurs and natural activation is prevented
Solution Approach 1:
The patent uses a targeted integration system as an intermediary mechanism between the lentiviral vector and the B cell genome. Instead of random integration, the system employs specific integration sites within the immunoglobulin locus, allowing controlled antibody secretion while preserving natural B cell activation pathways. This mediator approach resolves the contradiction by enabling productivity through targeted integration while maintaining reliability through preserved natural activation mechanisms.
2Productivity
If transposons are used for genetic engineering of B cells, then antibody secretion is achieved, but promiscuous integration occurs and natural activation is prevented
Solution Approach 1:
The patent replaces the promiscuous transposon system with a targeted integration system that acts as an intermediary. The system uses specific integration sites and controlled mechanisms to insert transgenic B cell receptor sequences, enabling antibody secretion while avoiding random integration. This preserves the natural activation processes, resolving the contradiction between productivity and reliability.
3Productivity
If constitutive expression is used, then antibody production is achieved, but antigen-induced activation and affinity maturation are prevented
Solution Approach 1:
The patent transitions from static constitutive expression to dynamic antigen-induced expression. The transgenic B cell receptor is designed to be expressed only upon antigen binding, mimicking natural B cell activation. This dynamic system allows the B cells to undergo affinity maturation and class switch recombination in response to antigen exposure, resolving the contradiction between maintaining productivity and enabling adaptability.
Solution Approach 2:
The patent changes the expression parameter from constitutive to inducible. By designing the transgenic B cell receptor to respond to antigen binding, the system allows expression levels to change based on antigen presence. This parameter change enables both antibody production and affinity maturation, as the expression is now regulated by physiological signals rather than being fixed.
4Reliability
If repeated monoclonal antibody administration is used, then treatment effect is maintained, but financial cost increases and anti-drug antibody risk aggravates
Solution Approach 1:
The patent enables the patient's own B cells to produce the therapeutic antibody internally. By genetically engineering autologous B cells to express the desired B cell receptor, the body becomes self-sufficient in producing the treatment. This eliminates the need for repeated external administrations, reducing both financial cost and the risk of anti-drug antibody formation, while maintaining reliable treatment effects through endogenous production.
Data Source
AI summary
The present invention relates to methods and compositions for engineering B cells to express transgenic B cell receptor (BCR) for antigen-induced antibody secretion, compositions, methods and uses thereof in immunotherapy.