Viral Vector NK Cell Engineering for Stable Gene Transfer

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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 simple and efficient viral vector-based method for gene transfer into NK cells, utilizing lentiviral vectors, which includes culturing NK cells with cytokines and feeder cells, followed by transduction with viral particles, resulting in stable and robust long-term expression of transgenes.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If viral vector-based gene transfer method is used, then gene transfer rate and transgene expression stability are improved, but process complexity increases due to additional culturing and transduction steps

Engineering Contradiction:

Improvegene transfer rateVSAvoidprocess complexity

Core Design Contradiction:

Manufacturing precisionVSDevice complexity

Solution Approach 1:

NK cells are pre-activated with cytokines (IL-2, IL-15, IL-21) and optionally feeder cells before viral transduction to enhance their receptivity and transduction efficiency, ensuring high gene transfer rates while maintaining a systematic workflow

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

Viral vectors serve as intermediaries to deliver heterologous nucleic acids into NK cells, enabling efficient and stable transgene expression that cannot be achieved through direct transfection methods

Inventive Principle:

Principle #24Intermediary (Mediator)

2Productivity

If multiple cytokines and feeder cells are used during NK cell culturing, then NK cell activation and transduction efficiency are improved, but manufacturing cost and process time increase

Engineering Contradiction:

Improvetransduction efficiencyVSAvoidculture time

Core Design Contradiction:

ProductivityVSLoss of time

Solution Approach 1:

The culturing protocol allows dynamic adjustment of cytokine combinations (IL-2, IL-15, IL-21) and feeder cell types based on specific application requirements, enabling optimization of transduction efficiency while managing culture time and cost

Inventive Principle:

Principle #15Dynamics

Solution Approach 2:

Cytokine concentrations and combinations are optimized to achieve maximum NK cell activation and transduction efficiency within acceptable timeframes, balancing productivity with process duration

Inventive Principle:

Principle #35Parameter changes

Data Source

PatentUS12516292B2Methods of producing modified natural killer cells and methods of use

Publication Date: 2026.01.06 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.