Modified Natural Killer Cells: Viral Transduction After Cytokine Activation

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Solution Overview

Problem

Efficient genetic modification of primary NK cells for enhanced anti-tumor activity has been difficult to achieve, as many malignant cells express MHC class I antigens, making them resistant to lysis by autologous NK cells, and current gene transfer methods are cumbersome and inefficient.

Innovation Solution

A viral vector-based method for gene transfer into NK cells, utilizing lentiviral vectors, which allows for stable and long-term expression of transgenes, including cytokines and feeder cells to activate and expand the cells, resulting in high transduction efficiency and robust gene expression.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If viral vector-based gene transfer method is used, then transduction efficiency and gene expression stability are improved, but process complexity increases

Engineering Contradiction:

Improvegene expression stabilityVSAvoidprocess complexity

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

The patent applies preliminary action by activating NK cells with cytokines (IL-2, IL-15, IL-21) and feeder cells before viral transduction. This pre-activation prepares the cells to be more receptive to viral vector uptake, thereby improving transduction efficiency and gene expression stability without requiring complex post-transduction manipulation

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent uses feeder cells as an intermediary component to facilitate NK cell activation and maintenance during the gene transfer process. These feeder cells provide necessary growth factors and support signals, enabling efficient viral transduction and stable transgene expression while simplifying the overall process compared to direct transduction of resting NK cells

Inventive Principle:

Principle #24Intermediary (Mediator)

2Productivity

If cytokine activation and feeder cells are used, then NK cell expansion and transduction efficiency are improved, but culture time and resource requirements increase

Engineering Contradiction:

ImproveNK cell expansionVSAvoidculture time

Core Design Contradiction:

ProductivityVSLoss of time

Solution Approach 1:

The patent implements continuous useful action by maintaining NK cells in an activated state through continuous cytokine supplementation (IL-2, IL-15, IL-21) and feeder cell co-culture throughout the transduction and expansion process. This continuous activation ensures sustained high-level transduction efficiency and robust transgene expression, achieving rapid NK cell expansion within a optimized timeframe

Inventive Principle:

Principle #20Continuity of useful action

Solution Approach 2:

The patent applies universality by using a combination of cytokines (IL-2, IL-15, IL-21) that perform multiple functions simultaneously: promoting NK cell proliferation, enhancing metabolic activity, improving viral receptor expression, and supporting transgene expression. This multi-functional approach maximizes NK cell expansion efficiency without requiring sequential separate culture steps

Inventive Principle:

Principle #6Universality (Multi-functionality)

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 enables stable and long-term expression of transgenes in NK cells, enhancing their antitumor efficacy and potential for immunotherapy applications by producing large numbers of genetically reprogrammed NK cells.

Implementation Method 1

The population of activated NK cells are transduced with a viral vector including the one or more heterologous nucleic acids, for example by contacting the activated NK cells with viral particles including the viral vector

Methodology Applied

Scientific EffectViral transduction:

Implementation Method 2

The methods include culturing a population of isolated NK cells (such as NK cells isolated or purified from a subject) in the presence of one or more cytokines (for example, interleukin (IL)-2, IL-15, and/or IL-21) to produce a population of activated NK cells

Methodology Applied

Scientific EffectCytokine stimulation:

Data Source

PatentUS12448604B2Methods of producing modified natural killer cells and methods of use

Publication Date: 2025.10.21 THE GOVERNMENT OF THE UNITED STATES OF AMERICA AS REPRESENTED BY THE SECRETARY DEPARTMENT OF HEALTH & HUMAN SERVICES

AI summary

Disclosed herein are method of producing NK cells that include one or more heterologous nucleic acids. The methods include culturing a population of isolated NK cells in the presence of one or more cytokines to produce a population of activated NK cells. The population of activated NK cells are transduced with a viral vector comprising the one or more heterologous nucleic acids, for example by contacting the activated NK cells with viral particles including the viral vector. The resulting transduced NK cells are then cultured in the presence of one or more cytokines, and optionally in the presence of irradiated feeder cells, to produce a population of expanded transduced NK cells. Also disclosed are methods of treating a subject with a disorder (such as a tumor or hyperproliferative disorder) by administering to the subject NK cells produced by the methods described herein.