Cellular Reprogramming With Controlled OCT4/KLF4 for Safe Rejuvenation
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Solution Overview
Problem
Existing methods for cellular reprogramming, such as using the Yamanaka factors (OCT4, SOX2, c-Myc, and KLF4), often result in toxicities like teratoma formation and dysplasia, and fail to maintain cellular identity, limiting their applicability in vivo.
Innovation Solution
The use of spatially and temporally controlled expression of OCT4 and KLF4, without c-Myc, to rejuvenate cells by reversing epigenetic marks associated with aging, thereby restoring cellular identity and function without complete reprogramming.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If Yamanaka factors (OCT4, SOX2, c-Myc, and KLF4) are used for cellular reprogramming, then cellular rejuvenation and tissue regeneration are promoted, but toxicities such as teratoma formation and dysplasia occur
Solution Approach 1:
The patent extracts and removes c-Myc from the Yamanaka factor combination, using only OCT4, SOX2, and KLF4 for reprogramming. This extraction eliminates the primary source of tumorigenicity while preserving the rejuvenation effects, directly resolving the contradiction between efficacy and safety
Solution Approach 2:
The patent applies partial reprogramming by using a subset of the original four factors (three instead of four) and controlling the duration and intensity of factor expression. This partial action achieves sufficient rejuvenation without the excessive action that leads to teratoma formation
2Reliability
If complete reprogramming is performed to reverse aging, then cellular function is restored, but cellular identity is lost
Solution Approach 1:
The patent employs partial reprogramming with controlled expression of OCT4, SOX2, and KLF4 for limited time periods. This partial action reverses epigenetic aging marks and restores cellular function while avoiding complete dedifferentiation that would erase cellular identity
Solution Approach 2:
The patent uses dynamic, time-controlled expression of reprogramming factors rather than static continuous expression. The temporal dynamics allow reversible epigenetic changes that restore youthfulness without permanent loss of cellular identity
Data Source
AI summary
Provided herein are engineered nucleic acids (e.g., expression vectors, including viral vectors, such as lentiviral vectors, adenoviral vectors, AAV vectors, herpes viral vectors, and retroviral vectors) that encode OCT4; KLF4; SOX2; or any combination thereof that are useful, for example, in inducing cellular reprogramming, tissue repair, tissue regeneration, organ regeneration, reversing aging, or any combination thereof. Also provided herein are recombinant viruses (e.g., lentiviruses, alphaviruses, vaccinia viruses, adenoviruses, herpes viruses, retroviruses, or AAVs) comprising the engineered nucleic acids (e.g., engineered nucleic acids), engineered cells, compositions comprising the engineered nucleic acids, the recombinant viruses, engineered cells, engineered proteins, chemical agents that are capable of activating expression of OCT4; KLF4; SOX2; or any combination thereof, an engineered protein selected from the group consisting of OCT4; KLF4; SOX2; or any combination thereof, an antibody capable of activating expression of OCT4; KLF4; SOX2; or any combination thereof, and methods of treating a (e.g., ocular disease), preventing a disease (e.g., ocular disease), regulating (e.g., inducing or inducing and then stopping) cellular reprogramming, regulating tissue repair, regulating tissue regeneration, or any combination thereof).