iPS Cell Reprogramming via Activated Ras Pathway
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Solution Overview
Problem
The efficiency of establishing induced pluripotent stem (iPS) cells is low, particularly when using three factors (Oct3/4, Sox2, and Klf4) excluding c-Myc, which can lead to tumorigenesis, and the relationship between the Ras family and somatic cell reprogramming is not well understood.
Innovation Solution
Increasing the level of activated Ras family members and their target factors (PI3 kinase, RalGEF, Raf, AKT, Rheb, TCL1, and S6K) during nuclear reprogramming, using specific mutants and nucleic acids to activate signal transduction pathways such as the PI3 kinase and AKT pathways, to enhance iPS cell establishment efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Object-affected harmful factors
If three factors (Oct3/4, Sox2, and Klf4) excluding c-Myc are used for iPS cell production, then tumorigenesis risk is reduced, but establishment efficiency becomes extremely low
Solution Approach 1:
The invention changes the parameter of signal transduction activity by introducing activated Ras family members (GTP-bound forms) and their downstream effectors (PI3 kinase, AKT, Rheb, S6K). This parameter change enhances the reprogramming efficiency without requiring c-Myc, thereby maintaining low tumorigenesis risk while improving productivity
Solution Approach 2:
The invention uses activated Ras family members and their target factors as intermediary substances to mediate the reprogramming process. These intermediaries activate signaling pathways (PI3K-AKT-mTOR-S6K) that enhance iPS cell establishment efficiency, allowing the system to achieve high productivity without the harmful c-Myc factor
2Device complexity
If conventional reprogramming methods are used, then the process is simple, but the establishment efficiency remains low
Solution Approach 1:
The invention modifies the biochemical parameters of the reprogramming process by introducing activated signaling molecules (Ras-GTP, PI3 kinase, AKT, Rheb, S6K) to enhance establishment efficiency while maintaining relative procedural simplicity
Data Source
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AI summary
The present invention provides a method of improving the efficiency of establishment of induced pluripotent stem cell, including increasing, in a nuclear reprogramming step of somatic cell, the level of activated form of one or more proteins selected from the group consisting of Ras family members, PI3 kinase, RalGEF, Raf, AKT family members, Rheb, TCL1 and S6K. The present invention also provides an agent for improving the efficiency of establishment of induced pluripotent stem cell, containing a factor selected from the group consisting of Ras family members, PI3 kinase, RalGEF, Raf, AKT family members, Rheb, TCL1 and S6K, and nucleic acids encode the same. Furthermore, the present invention provides a method of producing induced pluripotent stem cells, including contacting a somatic cell with a nuclear reprogramming substance and one or more factors selected from the group consisting of Ras family members, PI3 kinase, RalGEF, Raf, AKT family members, Rheb, TCL1 and S6K, and nucleic acids that encode the same.