Partial Cellular Reprogramming Without c-Myc 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), result in toxicities like teratoma formation and dysplasia, and fail to maintain cellular identity, limiting their application in vivo.
Innovation Solution
Spatially and temporally controlled expression of OCT4 and SOX2, with or without KLF4, in the absence of c-Myc, to rejuvenate cells by reversing epigenetic marks associated with aging without complete reprogramming, using inducible promoters and AAV delivery.
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 epigenetic reversal are achieved, but teratoma formation and acute toxicities occur
Solution Approach 1:
The patent extracts and removes c-Myc from the Yamanaka factor combination, using only OCT4, SOX2, and KLF4. This extraction eliminates the primary source of teratoma formation and acute toxicities while preserving the cellular reprogramming and epigenetic reversal functions, as demonstrated by the safe application in both in vitro and in vivo models
Solution Approach 2:
The patent applies partial reprogramming by using a subset of the original Yamanaka factors (three out of four) and controlling the duration and intensity of expression. This partial action achieves sufficient epigenetic reversal and cellular rejuvenation without the excessive action that would lead to complete pluripotency and subsequent teratoma formation
2Reliability
If complete reprogramming is performed to reverse aging, then epigenetic marks are reset, but cellular identity is lost
Solution Approach 1:
The patent implements partial reprogramming by using controlled expression of OCT4, SOX2, and KLF4 that is sufficient to reverse epigenetic marks associated with aging but insufficient to completely reprogram the cell to a pluripotent state. This maintains cellular identity while achieving epigenetic youthfulness
Solution Approach 2:
The patent employs periodic or transient expression of the reprogramming factors rather than continuous expression. This temporal control allows epigenetic marks to be reset during specific periods while preventing permanent loss of cellular identity, as the factors are removed before complete reprogramming can occur
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).