NK Cells Engineered to Eliminate Cbl-b for Enhanced Tumor Targeting
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Solution Overview
Problem
Natural Killer (NK) cells are challenging to work with in immunotherapy due to difficulties in obtaining sufficient numbers for adoptive transfer and maintaining their tumor-targeting capabilities during culture and expansion, necessitating improved functions for effective targeting and elimination of tumor cells and virally infected cells.
Innovation Solution
Development of human NK cells lacking Cbl-b expression, either completely (Cbl-bneg) or with reduced expression (Cbl-blow), which are genetically modified to express a chimeric antigen receptor (CAR) targeted to cancer antigens, and activated through culture with feeder cells in the presence of cytokines like IL-15 and IL-2.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If NK cells are cultured and expanded to obtain sufficient numbers for adoptive transfer, then the quantity of NK cells increases, but their tumor-targeting capabilities are lost
Solution Approach 1:
The patent extracts and removes the Cbl-b protein, which acts as a negative regulator of NK cell function. By eliminating this inhibitory component through genetic modification (CRISPR/Cas9-mediated knockout or shRNA suppression), the NK cells maintain their tumor-targeting capabilities while allowing for expansion. The removal of Cbl-b prevents the loss of function that normally occurs during culture expansion.
Solution Approach 2:
The patent changes the molecular parameter of Cbl-b expression levels in NK cells. By reducing or eliminating Cbl-b expression through genetic modification, the NK cells undergo a functional transformation that allows them to maintain tumoricidal activity during expansion. This parameter change (from normal Cbl-b expression to reduced/eliminated expression) enables both high numbers and high functionality to coexist.
2Manufacturing precision
If NK cells are genetically modified to express CAR, then their specificity for cancer antigens improves, but the complexity of the cell preparation process increases
Solution Approach 1:
The patent combines two genetic modifications in a single NK cell population: CAR expression for antigen specificity and Cbl-b knockout for functional enhancement. This merging of modifications into one unified cell product simplifies the overall preparation process compared to separate steps, while achieving both high specificity and high functionality simultaneously.
3Power
If Cbl-b expression is reduced or eliminated in NK cells, then cytotoxicity and IFN-γ secretion increase, but the risk of autoimmunity may increase
Solution Approach 1:
The patent uses the CAR construct as an intermediary mechanism that provides antigen-specific direction to the enhanced NK cell cytotoxicity. The CAR ensures that the increased killing power is targeted specifically at cancer cells expressing the relevant antigen, rather than indiscriminately affecting normal cells. This intermediary (CAR) mediates between the enhanced function (from Cbl-b removal) and selective targeting, reducing off-target effects.
Data Source
AI summary
Described herein are compositions comprising human natural killer (NK) cells engineered to reduce or eliminate Cbl-b and with or without a chimeric antigen receptor (CAR), methods of making such compositions, and methods of using such compositions (e.g., killing cancer cells, treating a subject having cancer or viral infection).