NK Cell Cryopreservation with Kinase-Inhibitor Deactivation
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Solution Overview
Problem
Current methods of cryopreservation for NK cells, used in off-the-shelf immunotherapy, fail to maintain cell functionality and viability post-thawing, particularly affecting their cytotoxicity and expansion capabilities, necessitating the use of fresh cells in clinical trials.
Innovation Solution
Deactivating NK cells with kinase inhibitors like rapamycin or Dasatinib before cryopreservation, followed by appropriate thawing and reactivation, to enhance survival and cytotoxicity post-thawing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If NK cells are cryopreserved using current methodologies, then cell storage is enabled for off-the-shelf immunotherapy, but cell functionality and viability are compromised post-thawing
Solution Approach 1:
The patent applies preliminary action by deactivating NK cells before cryopreservation through treatment with deactivating agents (such as kinase inhibitors or mTOR inhibitors). This pre-treatment prepares the cells for successful cryopreservation by reducing their metabolic activity and preventing activation-induced damage during storage, thereby maintaining functionality and viability post-thawing while enabling extended shelf life for off-the-shelf immunotherapy products
2Productivity
If NK cells are activated and expanded for therapy, then therapeutic efficacy is improved, but cells become sensitive to cryopreservation damage
Solution Approach 1:
The patent applies the inversion principle by reversing the conventional sequence of operations. Instead of activating and expanding NK cells immediately for therapy (which makes them vulnerable to cryopreservation damage), the method first deactivates the cells, then cryopreserves them, and only reactivates them after thawing. This inverted approach allows therapeutic efficacy to be maintained while protecting against cryopreservation sensitivity through the intermediate deactivation step
3Reliability
If fresh NK cells are used in clinical trials, then cell functionality is maintained, but time consumption and cost increase
Solution Approach 1:
The patent applies preliminary action by deactivating NK cells before cryopreservation, which enables the cells to be stored for extended periods while maintaining functionality. This allows clinical trials to use thawed cells from inventory rather than requiring fresh cells, thereby reducing time consumption and cost while preserving cell functionality through the deactivation-cryopreservation-reactivation protocol
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 NK cells with improved viability and cytotoxicity post-thawing, allowing for effective off-the-shelf use and prolonged shelf life, comparable to fresh cells.
Implementation Method 1
Deactivating NK cells with kinase inhibitors like rapamycin or Dasatinib before cryopreservation
Implementation Method 2
cryopreserved, and subsequently thawed as described herein
Data Source
AI summary
Provided herein are methods for deactivation of effector cells, such as NK cells, including cells for adoptive cell therapy that are off-the-shelf cells. The deactivated cells may be cryopreserved following deactivation. The deactivated and cell may be expanded and/or activated prior to deactivation, and may or may not comprise a transgene, which may optionally encode a chimeric antigen receptor and/or T cell receptor. In certain embodiments, disclosed are methods for deactivation of NK cells comprising treatment with a kinase inhibitor, for example an mTOR inhibitor and/or a tyrosine kinase inhibitor, for example rapamycin and/or Dasatinib. Also provided herein are deactivated and cryopreserved NK cells, and methods of preparing and/or using the same for treatment of a subject in need thereof, for example treatment of a subject with cancer.