Chromatin Modifying Enzyme Inhibition for Stem Cell Reprogramming

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Solution Overview

Problem

Current methods for producing induced pluripotent stem cells are inefficient and can lead to oncogene upregulation, increasing the risk of tumor formation due to the use of multiple transcription factors with unwanted side effects.

Innovation Solution

Inhibiting Dot1L, YY1, and SUV39H1 enzymes during the reprogramming process to enhance efficiency and reduce the number of transcription factors required, using enzyme inhibitors or RNAi molecules to knock down their expression, and culturing cells under specific reprogramming conditions with a cocktail containing Oct4 and Sox2, potentially omitting Klf4 and c-Myc.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If multiple transcription factors (including Klf4 and c-Myc) are used for reprogramming, then reprogramming efficiency is improved, but the risk of oncogene upregulation and tumor formation increases

Engineering Contradiction:
Improvereprogramming efficiencyVSAvoidoncogene upregulation and tumor formation risk
Core Design Contradiction:
ProductivityVSObject-affected harmful factors

Solution Approach 1:

The patent extracts and removes the harmful transcription factors Klf4 and c-Myc from the reprogramming cocktail while retaining the essential factors Oct4 and Sox2. This selective removal eliminates the oncogenic risk associated with Klf4 and c-Myc while maintaining reprogramming functionality through the action of Dot1L inhibitor and YY1 inhibitor that compensate for their absence.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent converts the harmful effect of Dot1L and YY1 (which maintain differentiated state and inhibit reprogramming) into a benefit by using specific inhibitors against these enzymes. The Dot1L inhibitor and YY1 inhibitor transform these obstacles into facilitators of reprogramming, enabling efficient differentiation without requiring oncogenic transcription factors.

Inventive Principle:
Principle #22Blessing in disguise (Convert harm into benefit)

2Productivity

If conventional reprogramming methods are used, then stem cells can be produced, but the process is time-consuming and inefficient

Engineering Contradiction:
Improvestem cell production efficiencyVSAvoidreprogramming time
Core Design Contradiction:
ProductivityVSLoss of time

Solution Approach 1:

The patent applies preliminary inhibition of Dot1L and YY1 enzymes before initiating the reprogramming process with Oct4 and Sox2. This pre-treatment prepares the cellular environment by removing epigenetic barriers in advance, allowing the reprogramming to proceed more rapidly and efficiently without the time delays associated with conventional methods.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The patent changes the biochemical parameters of the reprogramming system by introducing Dot1L inhibitor and YY1 inhibitor at specific concentrations and timing. These parameter modifications create optimal conditions for rapid reprogramming, reducing the time required for cellular differentiation while maintaining high efficiency through controlled enzymatic inhibition.

Inventive Principle:
Principle #35Parameter changes

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

This approach accelerates and simplifies the reprogramming process, reducing the risk of oncogenesis by using fewer oncogenic transcription factors and increasing the yield of induced pluripotent stem cells, allowing for more efficient production of pluripotent stem cells that can differentiate into various cell types.

Implementation Method 1

inhibiting Dot1L (a histone H3 methyltransferase), and one or more of YY1 (a transcriptional repressor protein), and/or SUV39H1 (a histone-lysine N-methyltransferase) helps promote the reprogramming of differentiated cells

Methodology Applied
Scientific EffectHistone methylation:

Implementation Method 2

Knock down of Dot1L, YY1, and/or SUV39H1 expression in a differentiated cell can be achieved by any appropriate technique, including, for example, by using RNAi (e.g., siRNA, shRNA) molecules

Methodology Applied
Scientific EffectRNA interference:

Data Source

PatentEP2766474B1Inhibition and enhancement of reprogramming by chromatin modifying enzymes
Publication Date: 2020.10.07 CHILDRENS MEDICAL CENT CORP
  • EP2766474B1 patent drawingFigure 1A~1D
  • EP2766474B1 patent drawingFigure 2B~2E
  • EP2766474B1 patent drawingFigure 3A~3E

AI summary

Methods and compositions are provided for the production of stem cells and induced pluripotent stem cells, and for uses thereof.