Genetically Modified MSCs for Stable Extracellular Vesicle Production

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Solution Overview

Problem

Current MSC-based therapies face challenges such as limited therapeutic efficacy in clinical trials, high doses required for effect, and the need for frequent donor changes due to MSC senescence, leading to increased costs and inconsistent results.

Innovation Solution

Genetically modifying MSCs to overexpress HIF-1α and hTERT, resulting in EVs with enhanced therapeutic potency, which are then cultured in a cytokine-based medium to increase immunosuppressive capacity and stability, providing a constant source of boosted immunosuppressive EVs.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If MSCs are used for therapeutic strategies, then immunosuppressive capacity and tissue regeneration are improved, but MSC senescence occurs leading to frequent donor changes and increased costs

Engineering Contradiction:
Improvetherapeutic efficacyVSAvoidMSC lifespan
Core Design Contradiction:
ReliabilityVSDuration of action of stationary object

Solution Approach 1:

The patent applies preliminary action by pre-modifying MSCs with hTERT and HIF-1α before use to prevent senescence and enhance therapeutic efficacy. The genetic modifications are performed in advance during cell culture, allowing the cells to be prepared with extended lifespan and improved function before clinical application, thus avoiding the need for frequent donor changes.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The patent changes key parameters of MSCs through genetic modification. hTERT overexpression alters the cellular aging parameter by maintaining telomere length, while HIF-1α overexpression modifies the therapeutic efficacy parameter by enhancing immunosuppressive capacity and tissue regeneration potential, directly addressing both contradictions.

Inventive Principle:
Principle #35Parameter changes

2Reliability

If high doses of MSCs are administered, then therapeutic effect is improved, but production costs and complexity increase

Engineering Contradiction:
Improvetherapeutic effectVSAvoidproduction complexity
Core Design Contradiction:
ReliabilityVSDevice complexity

Solution Approach 1:

The patent changes the functional parameters of MSCs through genetic modification. By overexpressing HIF-1α, the cells acquire enhanced immunosuppressive and tissue-regenerative capabilities at the cellular level, which translates to improved therapeutic effects at lower doses, thereby reducing production complexity and costs.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent applies preliminary action by pre-enhancing the therapeutic capacity of MSCs through genetic modification and cytokine conditioning before administration. This preliminary enhancement allows lower doses to achieve the same therapeutic effect, simplifying production requirements.

Inventive Principle:
Principle #10Preliminary action

3Reliability

If MSCs are genetically modified to overexpress HIF-1α and hTERT, then therapeutic potency and stability are improved, but manufacturing complexity increases

Engineering Contradiction:
Improvetherapeutic potencyVSAvoidmanufacturing ease
Core Design Contradiction:
ReliabilityVSEase of manufacture

Solution Approach 1:

The patent applies preliminary action by performing genetic modification and cytokine conditioning during the cell culture phase before clinical use. The lentiviral transduction and cytokine treatment are integrated into the existing MSC production workflow, allowing modifications to be made in advance when cells are being cultured and expanded, thus minimizing impact on manufacturing ease.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The patent makes the manufacturing process multi-functional by combining genetic modification, cytokine conditioning, and cell expansion in a unified workflow. The same cell culture system used for MSC production is also used for introducing genetic modifications and conditioning, eliminating the need for separate complex manufacturing 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 modified EVs demonstrate increased immunosuppressive capacity, reduced senescence, and improved therapeutic potential, effectively treating autoimmune and ischemic diseases with enhanced stability and reduced production costs.

Implementation Method 1

transduced with a lentiviral vector containing the human telomerase enzyme (hTERT) and hypoxia-inducible factor 1 alpha (HIF-1α) genes

Methodology Applied
Scientific EffectViral transduction:

Implementation Method 2

The EVs were isolated from mesenchymal stromal cells genetically modified to overexpress HIF-1α and hTERT

Methodology Applied
Scientific EffectGene integration:

Implementation Method 3

overexpression of hTERT generates the unlimited and constant division of the cells, giving rise to a stable and homogeneous source of vesicles

Methodology Applied
Scientific EffectTelomerase activity:

Implementation Method 4

overexpression of HIF-1α gives rise to a greater therapeutic potential

Methodology Applied
Scientific EffectHypoxia-inducible factor signaling:

Implementation Method 5

Mesenchymal stromal cells (MSCs) are multipotent progenitor cells... capable to differentiate into the chondrocyte, osteoblast or adipocyte lineages

Methodology Applied
Scientific EffectExtracellular vesicle secretion:

Data Source

PatentEP4023231A1Extracellular vesicles derived from mesenchymal stromal cells genetically modified to overexpress HIF-1a and htert
Publication Date: 2022.07.06 FUNDACION PARA LA INVESTIGACION HOSPITAL UNIVERSITARIO Y POLITECNICO LA FE DE LA COMUNIDAD VALENCIANA
  • EP4023231A1 patent drawingFigure 1A~1D
  • EP4023231A1 patent drawingFigure 1E~1G
  • EP4023231A1 patent drawingFigure 2A~2C

AI summary

The present invention refers to extracellular vesicles (EVs) isolated from mesenchymal stromal cells genetically modified to overexpress HIF-1α and hTERT (MSCs-T-HIF). The present invention also refers to the method for obtaining the extracellular vesicles. Finally, the clinical applications of the EVs in cellular therapy, in particular for the treatment of autoimmune diseases and ischemic diseases, are also contemplated.