Sequential TALEN Double Knockdown in TILs to Limit Translocation Risk
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for treating bulky, refractory cancers using adoptive autologous transfer of tumor infiltrating lymphocytes (TILs) are limited in effectiveness, and there is a need for additional treatment approaches, particularly in enhancing the therapeutic response of TILs through gene-editing techniques to address chromosomal translocation risks.
Innovation Solution
A sequential double KO process using TALEN systems is employed to target and reduce the expression of PD-1 and TIGIT in TILs, involving a spaced-out delivery of TALEN systems to edit the genes, followed by specific culture conditions to enhance TIL therapeutic efficacy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If TALEN systems are delivered to edit PD-1 and TIGIT genes in TILs, then therapeutic effect of TILs is enhanced, but risk of chromosomal translocations increases
Solution Approach 1:
The patent divides the gene editing process into two separate sequential steps: first delivering TALEN to knockout PD-1, then after a resting period, delivering TALEN to knockout TIGIT. This segmentation of the editing process into spaced-out deliveries reduces chromosomal translocation risk while achieving dual knockout of both genes to enhance therapeutic effect
Solution Approach 2:
The patent implements a resting period between the first and second TALEN deliveries, allowing the cell population to recover and reducing cumulative stress on the cells. This preliminary recovery action before the second editing step helps minimize chromosomal translocation risk while maintaining editing efficacy
2Reliability
If sequential double KO process is implemented, then PD-1 and TIGIT expression is reduced, but process complexity increases
Solution Approach 1:
The patent segments the complex dual gene knockout into two simpler sequential steps, each targeting one gene at a time. This segmentation makes the process more manageable and controllable compared to attempting simultaneous dual knockout, reducing overall process complexity while achieving the desired gene expression reduction
Solution Approach 2:
The patent employs periodic action by alternating between editing phases and resting phases. The cycle of deliver TALEN → rest → deliver TALEN creates a rhythmic process that allows monitoring and control at each stage, making the complex dual knockout process more systematic and easier to manage
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
The method results in a population of TILs with reduced PD-1 and TIGIT expression, enhancing their therapeutic effect and potentially improving cancer treatment outcomes.
Implementation Method 1
introducing a first TALE nuclease (TALEN) system targeting a first gene selected from the group consisting of a gene encoding PD-1 and a gene encoding TIGIT into at least a portion of the third population of TILs
Data Source
AI summary
The present invention provides methods for preparing expanded tumor infiltrating lymphocytes (TILs) having reduced expression of PD-1 and TIGIT using sequential electroporation of two TALEN systems targeting PD-1 and TIGIT. Such TILs find use in therapeutic treatment regimens for cancer patients.