CN-121975740-A - Alternative reprogramming factor combinations and methods and uses thereof for inducing pluripotent stem cells

Abstract

The invention relates to the field of biotechnology, in particular to an alternative reprogramming factor combination and a method and application thereof for inducing pluripotent stem cells. The reprogramming factor combination comprises OCT4, c-MYC and RBBP5, reduces the number of insertion factors, improves the stability of genome, and simultaneously remarkably improves the induction efficiency of iPSCs, thereby having good industrialization prospect in the fields of regenerative medicine, disease modeling, drug screening, cell therapy and the like.

Inventors

• JIANG ZHENLONG
• Yang Songqin
• WANG ERKANG
• WANG JIN

Assignees

• 中国科学院长春应用化学研究所

Dates

Publication Date

20260505

Application Date

20260210

Claims (10)

1. Use of rbbp5 transcription factor in the preparation of a formulation for reprogramming somatic cells to induced pluripotent stem cells.
2. 2. A transcription factor composition comprising an RBBP5 transcription factor, an OCT4 transcription factor, and a c-MYC transcription factor.
3. 3. A nucleic acid composition comprising at least one of a nucleic acid encoding an RBBP5 transcription factor, a nucleic acid encoding an OCT4 transcription factor, or a nucleic acid encoding a c-MYC transcription factor.
4. 4. The biological material is characterized by comprising at least one of the following A) to D): a) A plasmid vector comprising at least one selected from a nucleic acid encoding an RBBP5 transcription factor, a nucleic acid encoding an OCT4 transcription factor, or a nucleic acid encoding a c-MYC transcription factor; B) A plasmid combination comprising a helper plasmid and a plasmid vector as described in a); C) Transforming and/or transfecting a host cell with the plasmid vector as defined in A) or the plasmid combination as defined in B); D) And a viral vector produced by the packaging of the host cell of C).
5. 5. The biomaterial of claim 4, wherein the helper plasmid comprises a packaging plasmid and/or an envelope plasmid; the viral vector comprises at least one of a retroviral vector, an adenoviral vector, an adeno-associated viral vector, a baculovirus vector, an alphaviral vector, a picornaviral vector or a lentiviral vector.
6. 6. Use of the transcription factor composition of claim 2, the nucleic acid composition of claim 3, or the biological material of claim 4 or 5 in the preparation of a reagent or kit for reprogramming somatic cells to induced pluripotent stem cells.
7. 7. An agent or kit for reprogramming a somatic cell to induce a pluripotent stem cell, comprising at least one of the transcription factor composition of claim 2, the nucleic acid composition of claim 3, or the biological material of claim 4 or 5.
8. 8. A method of reprogramming a somatic cell to an induced pluripotent stem cell comprising: contacting a somatic cell with the reagent or kit of claim 7, and culturing to obtain the induced pluripotent stem cell.
9. 9. An induced pluripotent stem cell, prepared by the method of claim 8.
10. 10. Use of the induced multifunctional stem cells of claim 9 for the preparation of a product for regenerative medicine, disease modeling, drug screening or cell therapy.

Description

Alternative reprogramming factor combinations and methods and uses thereof for inducing pluripotent stem cells Technical Field The invention relates to the field of biotechnology, in particular to an alternative reprogramming factor combination and a method and application thereof for inducing pluripotent stem cells. Background Induced Pluripotent Stem Cells (iPSCs) are cell populations in which somatic cells are reprogrammed to a dedifferentiated state by specific gene expression, have multipotency highly similar to Embryonic Stem Cells (ESCs), can differentiate into almost all types of cells in vivo, show wide application prospects in various fields of regenerative medicine, disease model construction, drug screening, toxicity testing and the like, and provide important cell resources for research and transformation application in related fields. Currently, the most classical induction method is to introduce the four "mountain factors" OCT4, SOX2, KLF4 and c-MYC (OSKM) into cells via viral vectors. Although effective, the technology has two major bottlenecks, namely low reprogramming efficiency and random integration of a plurality of exogenous genes mediated by viruses, which can obviously increase genome instability and cancerogenic risk and endanger the safety of clinical application. In order to improve the induction efficiency, other factors (such as KDM 1B) are added on the basis of OSKM in the prior art, but the integration quantity of the exogenous genes is further increased, so that the fundamental safety problem cannot be solved, and the risk of genome variation is possibly aggravated. Therefore, it is always difficult for existing solutions to balance between "high efficiency" and "high safety". Recent studies have shown that epigenetic regulation plays a key role in cell reprogramming. RBBP5, as a core subunit of the MLL/SET1 methyltransferase complex, can be assembled with proteins such as WDR5, ASH2L, DPY and the like to form a functional complex, and specifically catalyzes mono-, di-and trimethylated modifications of histone H3K 4. The H3K4me3 is used as a marker modification of an active promoter, has key regulation and control effects on gene transcriptional activation and chromatin remodeling, and the biological processes are the core mechanism of cell fate transition in the process of reprogramming somatic cells into iPSCs, so that RBBP5 is suggested to have important potential value in an iPSCs-induced epigenetic regulation and control network. In summary, the technical scheme of inducing iPSCs by using the existing lentivirus is difficult to achieve both the induction efficiency and the genome stability, and development of a new technology capable of effectively improving the induction efficiency of the iPSCs while reducing the number of insertion factors and improving the genome stability is needed to be urgently developed so as to break through the bottleneck of the prior art and promote the wide application of the iPSCs in various fields. Disclosure of Invention In view of this, the technical problem underlying the present invention is to provide alternative reprogramming factor combinations. The factor combination can effectively improve the induction efficiency of iPSCs while reducing the number of exogenous insertion factors and improving the stability of cell genome, and realizes the cooperative optimization of reprogramming effect and cell safety. The invention provides application of RBBP5 transcription factor in preparing preparation for reprogramming somatic cells into induced pluripotent stem cells. In some embodiments, the function of the formulation includes at least one of reprogramming somatic cells to induce pluripotent stem cells, facilitating reprogramming by affecting an early-reprogramming EMT process via a tgfβ pathway, or enhancing expression of an early-reprogramming EMT process-related gene. The present invention provides transcription factor compositions comprising RBBP5 transcription factor, OCT4 transcription factor and c-MYC transcription factor. Compared with other reprogramming factor combinations, the composition can synergistically improve the reprogramming efficiency of cells, has excellent effects in reducing the integration risk of exogenous genes and maintaining the stability of cell genome, and realizes the double optimization of the efficiency and the safety of the reprogramming process. The invention also provides a nucleic acid composition comprising at least one of a nucleic acid encoding an RBBP5 transcription factor, a nucleic acid encoding an OCT4 transcription factor, or a nucleic acid encoding a c-MYC transcription factor. In some embodiments, the nucleic acid sequence encoding the RBBP5 transcription factor is NCBI reference transcript nm_005057, or a nucleic acid sequence having at least 80% identity to a nucleic acid sequence as set forth above; The nucleic acid sequence encoding the OCT4 transcription factor is NCBI reference transcri