Engineered Lymphocytes for Targeted Gene Editing Delivery
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Solution Overview
Problem
Current methods for in vivo delivery of gene editing reagents to target cells face limitations such as immunogenicity, vector carrying capacity, and low delivery efficiency, particularly in selectively targeting specific cells without causing toxicity.
Innovation Solution
Genetically modified lymphocytes with reduced cytotoxicity, engineered to express cargo molecules like gene editing reagents, utilize the perforin-granzyme pathway for targeted delivery by mutating cytotoxic proteins and incorporating cargo into cytolytic granules for selective transfer to target cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If viral vectors (rAAV, lentiviruses) or engineered polymers are used for in vivo delivery of gene editing reagents, then delivery to target tissues is achieved, but immunogenicity, vector carrying capacity, and delivery efficiency are compromised
Solution Approach 1:
The patent uses engineered lymphocytes as intermediary carriers to deliver gene editing reagents. These lymphocytes serve as a biological mediator that can transport cargo molecules (including gene editing reagents) from the bloodstream into target cells through their cytotoxic granules, avoiding the direct use of viral vectors that trigger immunogenicity.
Solution Approach 2:
The patent employs a replication-deficient lentiviral vector to transfer the cargo molecule into the lymphocyte, creating a copy of the therapeutic payload within the lymphocyte's granules. This allows the lymphocyte to serve as a factory for producing and delivering the gene editing reagent without requiring the lymphocyte itself to be divided or replicated.
2Reliability
If viral vectors or polymers are used for in vivo delivery, then cargo delivery is achieved, but selectivity for specific target cells is reduced
Solution Approach 1:
The patent imparts specific qualities to the lymphocyte population through engineering. By introducing a chimeric antigen receptor (CAR) that recognizes a specific antigen (e.g., CD19), the lymphocytes are given localized specificity to target only cells expressing that antigen. This creates heterogeneity in the lymphocyte population's targeting capability, allowing precise selection of target cells.
Solution Approach 2:
The patent modifies the lymphocyte's surface properties by introducing a chimeric antigen receptor with specific binding characteristics. This changes the lymphocyte's parameter for cell recognition from non-specific to highly specific, enabling differentiation between target cells (expressing the CAR antigen) and non-target cells (not expressing the antigen), thereby achieving precise target cell selectivity.
3Productivity
If cytotoxic lymphocytes are used for cargo delivery, then delivery capability is achieved, but cytotoxicity causes harm to target cells
Solution Approach 1:
The patent converts the harmful cytotoxic granules of lymphocytes into beneficial delivery vehicles. By redirecting the granule release pathway, the cargo molecule (gene editing reagent) is delivered through the same perforin-granzyme mechanism that would normally cause cell death. The harmful cytotoxic pathway is thus transformed into a useful delivery mechanism, allowing cargo transfer without killing the target cell.
Solution Approach 2:
The patent introduces an intermediary mechanism (the chimeric antigen receptor and engineered granule contents) that mediates between the lymphocyte's cytotoxic potential and the need for non-lethal cargo delivery. The intermediary CAR system ensures the lymphocyte identifies and binds to target cells, while the engineered granule contents ensure only the desired cargo is delivered through the cytotoxic pathway, preventing actual cell death.
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 enables efficient, selective, and non-toxic delivery of gene editing reagents to specific cells, overcoming the limitations of existing methods by utilizing genetically modified lymphocytes to target and deliver cargo molecules through the perforin-induced membrane pores, reducing cytotoxicity and improving delivery efficiency.
Implementation Method 1
delivery of the one or more cargo molecules (e.g., gene editing reagents) to the target cell, preferably through perforin-induced membrane pores
Data Source
AI summary
The present invention provides genetically modified lymphocyte with reduced cytotoxicity. Also provided are methods of using these genetically modified lymphocytes to deliver cargo molecules to a target cell.