Engineered Treg Cryopreservation for CD62L Retention

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Solution Overview

Problem

Cryopreservation of regulatory T cells (Tregs) leads to a loss of CD62L expression, impairing their migratory and homing functions and suppressive capabilities, necessitating lengthy post-thaw culture and expansion steps that are impractical in clinical settings.

Innovation Solution

Transducing Tregs with a polynucleotide encoding FOXP3 prior to cryopreservation to maintain CD62L expression and immunosuppressive function, allowing immediate use post-thawing without additional culture or expansion.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If Tregs are cryopreserved, then cell storage and flexibility in therapy planning are improved, but CD62L expression is lost and migratory function is impaired

Engineering Contradiction:

Improveflexibility in therapy planningVSAvoidmigratory function

Core Design Contradiction:

Adaptability or versatilityVSReliability

Solution Approach 1:

The patent applies preliminary action by transducing Tregs with FOXP3 polynucleotides before cryopreservation. This pre-treatment ensures that CD62L expression is maintained during the freezing process, allowing the cells to retain migratory function while enabling flexible storage and therapy planning. The FOXP3 overexpression prior to freezing protects the cells from cryopreservation-induced loss of surface markers.

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent changes the biochemical parameter of FOXP3 expression levels in Tregs. By increasing FOXP3 polynucleotide and polypeptide levels through transduction, the cells are modified to maintain CD62L expression during cryopreservation. This parameter change in FOXP3 expression directly counteracts the loss of migratory function that would normally occur during freezing.

Inventive Principle:

Principle #35Parameter changes

2Productivity

If Tregs are cryopreserved, then cell banking and off-the-shelf therapy are enabled, but suppressive capacity is reduced

Engineering Contradiction:

Improvecell banking efficiencyVSAvoidsuppressive capacity

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent implements preliminary action by introducing FOXP3 polynucleotides into Tregs before cryopreservation. This pre-transduction ensures that when cells are frozen for banking, they maintain their suppressive capacity upon thawing. The preliminary FOXP3 overexpression protects against the loss of immunosuppressive function that would otherwise occur during the freezing and storage process.

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent applies beforehand cushioning by using FOXP3 overexpression as a protective measure against cryopreservation damage. The elevated FOXP3 levels act as a buffer that prevents the loss of CD62L and other surface markers during freezing, thereby cushioning the cells against the harmful effects of cryopreservation and maintaining their therapeutic efficacy.

Inventive Principle:

Principle #11Beforehand cushioning (Prior cushioning)

3Quantity of substance

If Tregs undergo lengthy culture and expansion, then adequate cell numbers are obtained, but cell quality and suppressive function deteriorate

Engineering Contradiction:

ImproveTreg cell numberVSAvoidcell quality

Core Design Contradiction:

Quantity of substanceVSReliability

Solution Approach 1:

The patent applies preliminary action by transducing Tregs with FOXP3 before cryopreservation, enabling the cells to be stored and used immediately without prolonged culture. This approach obtains adequate cell numbers through efficient expansion protocols while maintaining high cell quality by avoiding the detrimental effects of extended culture periods. The FOXP3 pre-transduction allows cells to retain their suppressive function ready for immediate infusion.

Inventive Principle:

Principle #10Preliminary action

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

Preserves Treg immunosuppressive function and migratory ability, enabling immediate clinical use of cryopreserved Tregs with enhanced efficacy and safety for various therapeutic applications.

Implementation Method 1

introducing a polynucleotide encoding FOXP3 into a Treg or Treg population... FOXP3 expression in Tregs prior to cryopreservation

Methodology Applied

Scientific EffectGene expression:

Implementation Method 2

cryopreservation of therapeutic Tregs... freezing and thawing on the Treg cell population

Methodology Applied

Scientific EffectCryopreservation: Cryogenics

Data Source

PatentEP4729609A2Method for cryopreserving engineered tregs

Publication Date: 2026.04.22 QUELL THERAPEUTICS LTD

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AI summary

The present invention relates to a method of preserving CD62L expression in a regulatory T cell (Treg) population that has been cryopreserved, comprising introducing a polynucleotide encoding a FOXP3 polypeptide into the Treg population prior to cryopreservation. The present invention also relates to a method of preserving CD62L in a Treg population after cryopreservation, comprising introducing a polynucleotide encoding a FOXP3 polypeptide into the Treg population and cryopreserving said Treg population. Furthermore, the present invention relates to the use of an exogenous polynucleotide encoding FOXP3 for the preservation of CD62L expression in a Treg population after cryopreservation, and to a cryopreserved engineered Treg, pharmaceutical compositions comprising the cryopreserved engineered Treg and to therapeutic uses thereof.