Therapeutic NK Cell Expansion Through Ex Vivo Culture and Cryopreservation

Solution Overview

Problem

The efficacy of Natural Killer (NK) cell immunotherapy for cancer treatment is limited by the short-term persistence and impaired effector function of infused NK cells, due to insufficient numbers and limited proliferation and retention in the tumor microenvironment.

Innovation Solution

A method for culturing and expanding NK cells ex vivo using CD3-depleted fractions with irradiated CD3+ cells, nicotinamide, IL-15, and a CD3 agonist, followed by cryopreservation and thawing, to enhance homing, retention, and proliferation while maintaining killing activity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If NK cells are infused for cancer treatment, then therapeutic efficacy is improved, but persistence and effector function are limited due to short-term duration

Engineering Contradiction:

Improvetherapeutic efficacyVSAvoidpersistence duration

Core Design Contradiction:

ReliabilityVSDuration of action of moving object

Solution Approach 1:

The patent applies preliminary action by expanding NK cells ex vivo before infusion. The method involves culturing and expanding NK cells in vitro using specific conditions (including IL-15, CD3 agonist, and nicotinamide) to increase their numbers and enhance their persistence and effector function before administration to the patient, thereby resolving the contradiction between therapeutic efficacy and persistence duration

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent utilizes parameter changes by modifying culture conditions to optimize NK cell expansion. Specific parameters including cytokine concentrations (IL-15 at 10-100 U/mL), drug concentrations (nicotinamide at 0.1-10 mM), and cell density are adjusted to achieve enhanced persistence and effector function while maintaining therapeutic efficacy

Inventive Principle:

Principle #35Parameter changes

2Reliability

If NK cells are infused for cancer treatment, then therapeutic efficacy is improved, but effector function is impaired due to limited proliferation in vivo

Engineering Contradiction:

Improvetherapeutic efficacyVSAvoidproliferation capacity

Core Design Contradiction:

ReliabilityVSProductivity

Solution Approach 1:

The patent applies preliminary action by performing NK cell expansion ex vivo before infusion. The method involves culturing and expanding NK cells in vitro using specific conditions (including IL-15, CD3 agonist, and nicotinamide) to increase their numbers and enhance their persistence and effector function before administration to the patient, thereby resolving the contradiction between therapeutic efficacy and persistence duration

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent uses intermediaries (cytokines and drugs) to mediate NK cell expansion. IL-15, CD3 agonist, and nicotinamide act as intermediaries that stimulate NK cell proliferation and enhance their effector function in vitro, thereby improving therapeutic efficacy without requiring extensive in vivo proliferation

Inventive Principle:

Principle #24Intermediary (Mediator)

3Reliability

If NK cells are infused for cancer treatment, then therapeutic efficacy is improved, but homing and retention are limited due to insufficient numbers

Engineering Contradiction:

Improvetherapeutic efficacyVSAvoidNK cell number

Core Design Contradiction:

ReliabilityVSQuantity of substance

Solution Approach 1:

The patent applies preliminary action by performing NK cell expansion ex vivo before infusion. The method involves culturing and expanding NK cells in vitro using specific conditions (including IL-15, CD3 agonist, and nicotinamide) to increase their numbers and enhance their persistence and effector function before administration to the patient, thereby resolving the contradiction between therapeutic efficacy and persistence duration

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent utilizes parameter changes by modifying culture conditions to optimize NK cell expansion. Specific parameters including cytokine concentrations (IL-15 at 10-100 U/mL), drug concentrations (nicotinamide at 0.1-10 mM), and cell density are adjusted to achieve enhanced persistence and effector function while maintaining therapeutic efficacy

Inventive Principle:

Principle #35Parameter changes

Data Source

PatentUS20250283052A1Therapeutic NK cell populations

Publication Date: 2025.09.11 GAMIDA CELL

AI summary

Methods of expanding and cryopreserving a natural killer (NK) cell fraction for clinical use are provided, and particularly, methods for providing expanded, cryopreserved NK cell fractions and protocols for their use, which can be employed for applications in cell transplants and infusions for treatment of cancer and other disease.