CN-122005816-A - Application of targeting mesenchymal stem cell NAT10 and RNA acetylation in inflammatory diseases

Abstract

The invention belongs to the technical field of medicines, and particularly discloses application of targeted mesenchymal stem cells NAT10 and RNA acetylation in inflammatory diseases. The invention reveals for the first time that NAT10 mediated ac4C modification is a key positive regulatory factor for regulating the immune function of Mesenchymal Stem Cells (MSC), and inflammatory environment can inhibit the pathway to cause the weakening of MSC function. Based on this finding, the present invention upregulates NAT10 expression in MSCs by genetic means to prepare a functionally enhanced MSC. The cell shows higher ac4C acetylation level, and obviously promotes the expression of key immunosuppressive factors iNOS and COX2 and the secretion of products NO and PGE2, thereby obtaining stronger T cell inhibition capability. Animal experiments show that the MSC with enhanced functions has obvious and superior treatment effect compared with common MSC in disease models such as inflammatory liver injury, periodontitis and the like.

Inventors

• GU YUTING
• LU ERYI
• JIN MIN
• LIN LU
• YU WEIJUN

Assignees

• 上海交通大学医学院附属仁济医院

Dates

Publication Date

20260512

Application Date

20260224

Claims (10)

1. Application of NAT10 mediated ac4C modification as a regulation target for enhancing MSC immunosuppressive function and immunotherapeutic effect.
2. 2. The use according to claim 1, wherein the level of ac4C modification of the total RNA in the cells is increased by up-regulating the expression and/or activity of NAT10 in the MSCs, thereby enhancing the immunosuppressive function of the MSCs and enhancing the immunotherapeutic effect.
3. 3. A method of enhancing immunosuppressive function of an MSC, wherein the expression level and/or activity of NAT10 in the MSC is up-regulated.
4. 4. Mesenchymal stem cell with enhanced immunosuppressive function, characterized in that the expression level and/or activity of its endogenous NAT10 gene is artificially up-regulated by genetic modification.
5. 5. The mesenchymal stem cell of claim 4, wherein the functional enhancement is characterized by a higher ac4C modification level of total RNA than unmodified wild-type MSC, an ability to secrete immunosuppressive factors and their products under inflammatory factor stimulation, and an ability to inhibit T cell proliferation, both being stronger than wild-type MSC.
6. 6. The mesenchymal stem cell with enhanced immunosuppressive function according to claim 4, wherein the preparation method comprises introducing exogenous NAT10 encoding gene into MSC (mesenchymal stem cell) derived from bone marrow by genetic engineering technology to realize overexpression of NAT 10.
7. 7. A pharmaceutical composition comprising the immunosuppressive function-enhanced mesenchymal stem cell of any one of claims 4-6 and a pharmaceutically acceptable carrier or adjuvant.
8. 8. Use of an immunosuppressive function-enhanced mesenchymal stem cell of any one of claims 4-6 or a pharmaceutical composition of claim 7 for the preparation of a product for preventing and/or treating inflammatory diseases.
9. 9. The use according to claim 8, wherein the inflammatory disease comprises liver injury, periodontitis, enteritis, arthritis, pneumonia or graft versus host disease.
10. 10. The use according to claim 8, wherein the immunosuppressive function-enhanced mesenchymal stem cells are administered to the subject by systemic infusion or local injection.

Description

Application of targeting mesenchymal stem cell NAT10 and RNA acetylation in inflammatory diseases Technical Field The invention relates to the technical field of medicines, in particular to application of targeted mesenchymal stem cell NAT10 and RNA acetylation in inflammatory diseases. Background Mesenchymal Stem Cells (MSCs) exist in almost all tissues, and are not only capable of differentiating into osteoblasts, chondrocytes and adipocytes to support tissue regeneration, but also are attracting attention due to their strong immunomodulatory functions. MSCs are capable of modulating the activity of a variety of immune cells, including T cells, B cells, dendritic cells, macrophages and neutrophils, by secreting various immunosuppressive molecules, such as Nitric Oxide (NO) produced by Inducible Nitric Oxide Synthase (iNOS) and prostaglandin E2 (PGE 2) produced by cyclooxygenase-2 (COX 2). This ability establishes the therapeutic potential of MSCs in a range of immune-mediated inflammatory diseases such as graft versus host disease, multiple sclerosis, hepatitis, systemic lupus erythematosus, inflammatory bowel disease, and rheumatoid arthritis. The immunomodulatory function of MSCs is deeply affected by the inflammatory microenvironment. Preclinical and clinical evidence suggest that the combination of pro-inflammatory cytokines, particularly interferon-gamma (IFN-gamma), with tumor necrosis factor alpha (TNF-alpha) or interleukin-1 is critical for activating the immunosuppressive properties of MSCs by inducing immunosuppressive molecules. However, immunomodulation of MSCs is plastic and these pro-inflammatory signals also trigger responses that may impair MSC function. For example, inflammatory stimuli may up-regulate miR-155, UCHL1 and the like in MSC, thereby weakening the immunosuppressive effect, and the combination of IFN-gamma and TNF-alpha may promote MSC apoptosis, which may further impair the therapeutic potential. These complex and often conflicting effects emphasize the need to elucidate the precise mechanisms by which inflammatory signals regulate MSCs immunomodulation, which is critical to optimize their clinical therapeutic applications. Epigenetic modifications are thought to play a key role in MSC immunomodulation. RNA modification of the apparent transcriptome, including N6-methyladenosine, N7-methylguanosine, and N4-acetylcytidine (ac 4C), plays a key role in gene expression by regulating mRNA processing, such as splicing, transport, translation, and degradation. Among them, ac4C mediated by N-acetyltransferase 10 (NAT 10) is attracting increasing attention. NAT10 contains both an N-acetyltransferase domain and a nucleotide binding region, the only "writing" enzyme responsible for catalyzing ac4C formation. NAT10 mediated ac4C modification is involved in the pathogenesis of a variety of diseases including cancer, bone diseases and myocardial diseases. NAT10 mediated ac4C modification also participates in immune and inflammatory responses by modulating T cells, B cells, macrophages and neutrophils, and is involved in a range of immune related diseases such as sepsis, periodontitis and inflammatory bowel disease. These findings highlight the potential therapeutic targets for NAT10 and ac4C modifications as immune related diseases. However, the function and regulatory mechanisms of NAT10 and ac4C modifications in MSC immunomodulation remain largely unexplored. Although MSCs present great potential in the treatment of inflammatory diseases and the academic community is continually in going through the understanding of their immunomodulation mechanisms, the prior art and research still have several key shortcomings and limitations: (1) Insufficient cognition for negative effects on inflammatory microenvironments and lack of effective countermeasures. While the prior studies have recognized that the inflammatory environment has a "double sword" effect on MSC function, both activating and damaging its function, specific negative mechanisms such as miR-155 upregulation, autophagy activation, UCHL1 upregulation, and induction of apoptosis have been reported. However, these cognition are sporadic and incomplete. The most critical issue is the current lack of a ubiquitous strategy that can effectively counter or reverse these negative effects. Most approaches are limited to "suppressing" some negative factor, belonging to passive remediation, rather than actively enhancing the functional stability of the MSC from a more upstream, more fundamental level. (2) There is a significant gap in apparent transcriptomics studies, especially neglecting ac4C modifications. In the field of epigenetic regulation, current research is highly focused on DNA methylation and histone modification, and interest in emerging RNA modifications (apparent transcriptomes) is still in the beginning. For RNA modifications, especially NAT10 mediated ac4C modifications, the role in MSC immunomodulatory function is entirely a "b