Microfluidic TIL Modification via Constriction-Induced mRNA Uptake
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Solution Overview
Problem
Current Tumor Infiltrating Lymphocyte (TIL) therapies require patient lymphodepletion and high-dose IL-2 maintenance, which can be cumbersome and limit their applicability.
Innovation Solution
Modifying TILs to increase expression of co-stimulatory molecules and cytokines, such as CD86 and membrane-bound IL-2 or IL-12, through methods like passing cell suspensions through cell-deforming constrictions to facilitate nucleic acid uptake and expression.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If TILs are engineered ex vivo with mRNA to enhance potency, then tumor infiltration and immune activity are improved, but the complexity of the manufacturing process increases
Solution Approach 1:
The patent modifies physical parameters of the TILs by passing them through constrictions of specific dimensions (3.5-6 μm) to create controlled perturbations. This physical parameter change enables mRNA uptake without requiring complex viral vectors or chemical transfection methods, thereby reducing manufacturing complexity while maintaining engineering effectiveness
Solution Approach 2:
The patent extracts and delivers specific mRNA molecules (encoding cytokines, co-stimulatory molecules, or chimeric proteins) directly into TILs through the constriction method. This targeted extraction and delivery approach avoids the need for complex gene editing tools or viral vectors, simplifying the overall manufacturing process
2Productivity
If TILs are modified to increase co-stimulatory molecules and cytokines, then Granzyme B expression and proliferation are enhanced, but the risk of off-target effects increases
Solution Approach 1:
The patent applies local quality modification by introducing specific mRNA sequences that encode particular proteins (cytokines, co-stimulatory molecules, or chimeric proteins) only in the TILs that pass through the constriction. This localized molecular modification ensures that the enhanced Granzyme B expression and proliferation occur specifically in the desired cell population, minimizing off-target effects
Solution Approach 2:
The patent uses mRNA as an intermediary carrier to deliver genetic information into TILs. This intermediary approach allows for precise control over which proteins are expressed (by selecting specific mRNA sequences) and avoids the non-specific effects associated with direct protein transfer or genetic integration methods
3Reliability
If conventional TIL therapy is used, then clinical activity is achieved, but patient eligibility is limited due to requirement for lymphodepletion and high-dose IL-2
Solution Approach 1:
The patent performs preliminary action by engineering TILs ex vivo with mRNA encoding cytokines and co-stimulatory molecules before administration. This preliminary engineering equips the TILs with self-sustaining capabilities, eliminating the need for subsequent high-dose IL-2 maintenance and lymphodepletion protocols, thereby expanding patient eligibility
Solution Approach 2:
The patent enables self-service by introducing autonomous cytokine production capabilities into TILs through mRNA encoding. The engineered TILs can now produce their own growth factors and co-stimulatory signals, making them self-sustaining and independent of external cytokine support or lymphodepletion therapy, thus broadening patient eligibility
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
Enhances Granzyme B expression and proliferation of TILs upon activation, potentially allowing for broader patient eligibility, repeat dosing, and concurrent treatment with other therapies without the need for lymphodepletion or high-dose cytokine support.
Implementation Method 1
passing a cell suspension comprising the TILs through a cell-deforming constriction, thereby causing perturbations of the TILs such that a nucleic acid encoding the co-stimulatory molecule and/or a nucleic acid encoding the cytokine enters the TILs through the perturbations
Data Source
AI summary
The present application provides TILs comprising agents that enhance activity and/or proliferative capacity of the TILs, methods of manufacturing such TILs. and methods of using such modified TILs for enhancing an immune response.