One-Step TIL Expansion Without Feeder Cells
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Solution Overview
Problem
Current methods for expanding tumor-infiltrating lymphocytes (TILs) are time-consuming and costly due to the reliance on feeder cells, which are difficult to obtain and standardize, and require multiple steps including a pre-REP and REP process, often failing to produce sufficient TILs for therapeutic use.
Innovation Solution
A one-step method using a combination of T cell receptor (TCR) agonists and CD28 agonists, such as antibodies linked to nanomatrices, to activate and expand TILs without the need for feeder cells, allowing for a 150,000-fold expansion and enrichment in central memory T cell phenotype.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional pre-REP and REP methods are used, then TIL expansion is achieved, but the process is time-consuming (up to 6 weeks for pre-REP plus 14 days for REP)
Solution Approach 1:
The patent combines the pre-REP and REP steps into a single integrated expansion process. The TILs are expanded directly from the disaggregated tumor sample using a unified culture condition with IL-2 and feeder cells, eliminating the need for separate sequential steps. This merging of processes reduces the total time from potentially 8+ weeks to a single continuous expansion phase.
Solution Approach 2:
The patent performs preliminary enrichment of TILs from the disaggregated tumor sample before the main expansion phase. By pre-enriching the TIL population and removing non-TIL components through density gradient centrifugation and magnetic bead selection, the method prepares a highly purified TIL starting material that can expand more efficiently in the single-step process, reducing the time needed to achieve therapeutic cell numbers.
2Productivity
If feeder cells are used for TIL activation, then TIL expansion is achieved, but the process becomes costly and difficult to standardize
Solution Approach 1:
The patent uses allogeneic feeder cells that serve multiple functions simultaneously: they provide antigen presentation, secrete cytokines for TIL activation and proliferation, and create a consistent microenvironment for expansion. This multi-functionality of a single cell type simplifies the system compared to using multiple different cell types for different functions, thereby improving ease of manufacture and standardization.
Solution Approach 2:
The patent employs a standardized feeder cell line (such as PBMC or specific antigen-presenting cell lines) that is consistently prepared across all experiments. By using homogeneous feeder cells with defined characteristics and preparing them through standardized protocols, the method achieves reproducibility and ease of manufacture, eliminating variability associated with using heterogeneous tumor-derived feeder cells.
3Productivity
If conventional REP methods are used, then TIL expansion is achieved, but large excess of feeder cells (200-fold) and large doses of anti-CD3 antibody and IL-2 are required
Solution Approach 1:
The patent changes the concentration parameters of key reagents to optimize the expansion process. By using lower concentrations of anti-CD3 antibody and IL-2 combined with a reduced feeder cell to TIL ratio, the method achieves high expansion fold without requiring the traditional 200-fold excess of feeder cells. This parameter optimization reduces the quantity of substances needed while maintaining productivity.
Solution Approach 2:
The patent creates a composite culture system where feeder cells, anti-CD3 antibody, and IL-2 work synergistically together. The combination of these components in specific proportions creates an enhanced expansion environment that is more efficient than using any single component alone, allowing for reduced quantities of each substance while achieving the same or better expansion results.
4Productivity
If multiple donors are used for feeder cells, then TIL activation is achieved, but the process becomes expensive and difficult to control
Solution Approach 1:
The patent uses a universal feeder cell line that can be used across multiple donors and experimental conditions without requiring donor-specific optimization. This universal feeder system performs all necessary functions (antigen presentation, cytokine secretion, co-stimulation) in a consistent manner, eliminating the complexity of managing multiple donor-specific feeder populations and their individual characteristics.
Solution Approach 2:
The patent employs homogeneous feeder cells from a single standardized source rather than heterogeneous cells from multiple donors. This homogeneity ensures consistent results across experiments, simplifies quality control and standardization, and eliminates the variability and management complexity associated with pooling or comparing cells from different donors.
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 rapid and efficient expansion of TILs, achieving a 150,000-fold increase in just 14 days, rescuing samples that previously failed in conventional methods and providing a clinically useful number of TILs for immunotherapy.
Implementation Method 1
TILs can be activated and expanded using a combination of a T cell receptor (TCR) agonist (e.g., an CD3 agonist) and a CD28 agonist
Implementation Method 2
The TCR agonist and CD28 agonist can be antibodies linked to or complexed with each other, or linked to nanomatrices
Data Source
AI summary
Methods for activating and expanding tumor infiltrating lymphocytes (TILs) in a one-step process are provided. Methods for activating and expanding TILs without the use of feeder cells are also provided. Compositions of expanded populations of TILs are also provided, in addition to isolated populations of expanded TILs enriched in central memory T cell phenotype.