CISH-Edited Tumor-Infiltrating Lymphocytes for Faster TIL Manufacturing
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
There is a need for additional methods and compositions for CISH TALEN mediated gene inactivation/deletion for the treatment and/or prevention of cancer and inflammatory disorders, as well as the development of manufacturing processes for genetically modified tumor infiltrating lymphocytes (TILs) suitable for commercial scale manufacturing and regulatory approval.
Innovation Solution
A method of preparing genetically modified TILs with reduced expression of CISH and optionally PD-1 by introducing nucleic acids encoding Transcription activator-like effector nucleases (TALE-nucleases) to selectively inactivate the CISH gene, followed by expansion and culturing processes in controlled environments to produce a therapeutic population of TILs.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional TIL expansion methods are used, then the process is simple and quick, but the therapeutic efficacy is limited due to high CISH expression
Solution Approach 1:
The patent applies preliminary action by performing gene editing of the CISH gene in TILs before expansion and therapeutic administration. The TILs are genetically modified ex vivo to reduce CISH expression, then expanded and administered to patients. This preliminary genetic modification enables subsequent therapeutic efficacy improvement without complicating the overall treatment process.
Solution Approach 2:
The patent changes the genetic parameter of CISH expression in TILs by introducing nucleic acids encoding TALE-nucleases that selectively inactivate the CISH gene. This parameter change (reducing CISH expression from high to low) directly improves therapeutic efficacy by enhancing TIL functionality and reducing inhibitory signaling.
2Productivity
If TILs are expanded rapidly, then production time is reduced, but manufacturing precision and genetic modification efficiency decrease
Solution Approach 1:
The patent performs gene editing before expansion, allowing the TILs to be pre-modified with TALE-nucleases that will inactivate CISH during subsequent expansion. This sequencing enables rapid expansion while maintaining precision, as the genetic modification is established before the expansion process begins.
Solution Approach 2:
The patent replaces conventional mechanical expansion methods with a genetically enhanced approach using TALE-nucleases. This substitution allows for more precise control of TIL characteristics and CISH inactivation, improving manufacturing precision while maintaining or enhancing expansion rates through biological mechanisms rather than purely mechanical processes.
3Reliability
If CISH expression is reduced in TILs, then therapeutic efficacy improves, but the complexity of genetic modification increases
Solution Approach 1:
The patent uses TALE-nucleases as intermediary molecules to achieve CISH gene inactivation. These nucleases serve as mediators that deliver the genetic modification without requiring complex viral vectors or other sophisticated delivery systems. The TALE-nucleases bind to specific DNA sequences and facilitate precise cutting, enabling CISH reduction with relatively simple genetic modification technology.
Solution Approach 2:
The patent changes the CISH expression parameter from high to low by introducing nucleic acids encoding TALE-nucleases. This parameter change is achieved through a relatively straightforward genetic modification process compared to other gene editing approaches, thereby improving therapeutic efficacy without proportionally increasing modification complexity.
4Reliability
If TILs are cultured for extended periods, then polyclonality and efficacy improve, but production time and cost increase
Solution Approach 1:
The patent performs gene editing before culture, so that the TILs are pre-modified with CISH-inactivating TALE-nucleases. This allows for shorter culture durations while still achieving desired polyclonality and efficacy, as the genetic modification is established prior to the culture period rather than requiring extended culture to achieve modification.
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 produces a therapeutic population of TILs with reduced CISH and PD-1 expression, enhancing their efficacy for treating cancers such as melanoma, ovarian cancer, and other diseases, with improved interferon-gamma production and polyclonality.
Implementation Method 1
introducing into the TILs nucleic acid(s) encoding one or more first Transcription activator-like effector nucleases (TALE-nuclease) able to selectively inactivate by DNA cleavage a gene encoding CISH
Implementation Method 2
expanding the TILs
Implementation Method 3
producing a therapeutic population of TILs
Implementation Method 4
with improved interferon-gamma production and polyclonality
Data Source
AI summary
The present invention provides improved and/or shortened processes and methods for preparing TILs in order to prepare therapeutic populations of genetically modified TILs with reduced expression of CISH and optionally PD-1 as described herein.