CAR T Cell Manufacturing Without Expansion for Potent Cell Therapy
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Solution Overview
Problem
Existing processes for generating genetically engineered T cells, such as those expressing a chimeric antigen receptor (CAR), are time-consuming and may result in cell populations with low potency or persistence, necessitating improved manufacturing methods for cell therapy applications.
Innovation Solution
A method involving exposing primary T cells to a stimulatory reagent, introducing a viral vector encoding a recombinant protein, and harvesting the transformed cells within specific time frames to achieve a composition of engineered T cells with enhanced potency and stability, characterized by high percentages of naïve-like and central memory T cells, and controlled integrated vector copy numbers.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional processes are used to generate genetically engineered T cells, then the cells can be produced, but the process is time-consuming and results in cell populations with low potency or persistence
Solution Approach 1:
The patent applies preliminary action by stimulating T cells with anti-CD3 and anti-CD28 antibodies before viral vector transduction. This pre-stimulation step prepares the cells to receive and express the CAR construct more effectively, leading to higher potency engineered cells. The stimulation occurs before the actual engineering step, optimizing the cells for subsequent transformation and improving both speed and quality of the final product.
2Quantity of substance
If T cells are expanded before engineering, then more cells are available, but the process time increases and cell differentiation/exhaustion occurs
Solution Approach 1:
The patent uses viral vectors to directly transduce primary T cells, effectively copying the CAR construct into the cell genome without requiring extensive cell expansion. This direct transduction approach allows the cells to be engineered in their primary state, avoiding the time loss and differentiation issues associated with expanding cells before engineering. The viral vector acts as a template that delivers the CAR construct directly to the cell nucleus.
3Stability of the object's composition
If cells are harvested at later time points, then more stable integrated vector copy number is achieved, but the cells become more differentiated and exhausted
Solution Approach 1:
The patent implements feedback control by monitoring integrated vector copy number (iVCN) and harvesting cells at the optimal time point when iVCN is stable but before excessive differentiation occurs. The process checks for iVCN stability and uses this information to determine the precise harvesting timing, ensuring cells are collected at the peak of their engineering efficiency before potency is compromised by differentiation or exhaustion.
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 generates engineered T cells with improved potency and persistence, suitable for cell therapy, within a shortened timeframe compared to conventional methods, maintaining a high percentage of naïve-like and central memory T cells and controlled vector integration.
Implementation Method 1
introducing into T cells of the stimulated population, a viral vector comprising a heterologous polynucleotide encoding a recombinant protein, thereby generating a population of transformed cells
Data Source
AI summary
The present disclosure provides processes for genetically engineering T cells, such as primary CD4+ T cells and/or CD8+ T cells, for use in cell therapy that does not involve expanding the cells. In particular aspects, the provided processes successfully generate compositions of engineered T cells, such as containing populations of engineered T cells, that express a chimeric antigen receptor (CAR) within a shortened amount of time as compared to alternative engineering processes, such as processes that involve expanding the cells. In certain aspects, the provided processes successfully generate a composition of engineered T cells suitable for use in cell therapy within 4 days from when the process to stimulate or activate the cells is initiated. In some aspects, the resulting engineered cell compositions are composed of cell population that are less differentiated, less exhausted, and more potent than engineered T cell compositions generated by other means, such as by processes that involve expanding the cells. Also provided are compositions of T cells generated by the provided methods and their uses for treating subjects.