Application of type I classical dendritic cell in preparation of medicine for treating drug-induced hepatic failure

Abstract

The invention relates to an application of type I classical dendritic cells in preparation of drugs for treating drug-induced hepatic failure. The application focuses on the cell subtype cDC1 which is very important for balancing the immune homeostasis in the liver, the reduction of cDC1 subgroups is proved in the liver tissues of AILF patients and mice, and the feedback tolerance type cDC1 is fully proved to be capable of remarkably relieving liver injury and inflammatory cell infiltration in the liver tissues on the in-vivo level of an AILF mouse model, promoting liver regeneration and repair, and improving the liver injury and the inflammatory cell infiltration in the liver tissues. The process is related to the mechanism that P300 activates IDO1 gene transcription through mediating H3K27ac modification so as to promote the immune tolerance phenotype of cDC1, the tolerance type cDC1 further decomposes tryptophan and secretes kynurenine through high-expression IDO1, Tregs differentiation and functions are induced, the in-liver immune homeostasis is maintained, liver cell proliferation and damage repair are promoted by secreting immunosuppressive factors, and the immune tolerance phenotype of cDC1 is promoted. And an important early-stage experimental basis is provided for research and development of new drugs based on the targets.

Description

Technical Field

[0001] The present invention belongs to the field of drug-induced liver injury, and particularly relates to the application of type I classical dendritic cells in the preparation of drugs for treating drug-induced liver failure. Background Art

[0002] Drug-induced liver injury (DILI) is one of the main causes of acute liver failure. Acetaminophen (APAP) is one of the main pathogenic drugs and the most intensively studied hepatotoxic drug. The pathogenesis of APAP-induced acute liver failure (AILF) is complex. It is initiated by harmful metabolites produced by excessive APAP metabolism in the liver, involving multiple links such as oxidative stress, mitochondrial dysfunction and hepatocyte necrosis. In this process, abnormal activation of the host innate immune system and inflammatory cytokine storm are considered to be important driving factors for its progression. Therefore, in-depth exploration of the impact of the immune microenvironment on the pathogenesis and progression of AILF is of great significance for exploring effective prevention and treatment strategies.

[0003] Conventional dendritic cells (cDCs) are one of the main DC subsets in the liver. The cDC1 subtype is not only an important cell type for activating effector immunity, but also can promote the generation, stability and function of regulatory T cells (Tregs) through various mechanisms, playing an important role in regulating the balance between immune activation and immune tolerance, thereby maintaining liver immune homeostasis. Studies have reported that changes in the frequency of intrahepatic cDCs are associated with the severity of APAP-induced liver injury, but the specific role and mechanism of liver cDCs in AILF have not yet been elucidated. Therefore, exploring the specific role of cDCs in AILF and clarifying the important endogenous regulatory factors that maintain the immune tolerance phenotype of cDCs may help to discover new therapeutic targets for AILF. Summary of the invention

[0004] The present invention provides the use of type I classical dendritic cells in the preparation of a drug for treating drug-induced liver failure, aiming at the technical problem that existing drug intervention methods are insufficient.

[0005] The present invention provides the use of type I classical dendritic cells (cDC1) in preparing a drug for treating drug-induced liver failure.

[0006] Furthermore, the drug-induced liver failure includes acetaminophen-induced drug-induced liver failure.

[0007] Furthermore, the drug also contains pharmaceutically acceptable carriers and / or excipients.

[0008] Furthermore, the pharmaceutically acceptable carrier and / or excipient includes at least one of a diluent, a binder, a surfactant, a humectant, an adsorption carrier, a lubricant, a filler, and a disintegrant.

[0009] Beneficial Effects

[0010] The present invention focuses on the cell subtype cDC1, which is very important for balancing the immune homeostasis in the liver. The reduction of cDC1 subpopulation has been confirmed in the liver tissues of AILF patients and mice, and it has been fully confirmed in the in vivo level of the AILF mouse model that the reinfusion of tolerant cDC1 can significantly reduce liver damage and inflammatory cell infiltration in liver tissue, promote liver regeneration and repair, and this process is mechanistically related to P300 activating IDO1 gene transcription through H3K27ac modification to promote the immune tolerance phenotype of cDC1. The tolerant cDC1 further decomposes tryptophan to secrete kynurenine through the highly expressed IDO1, induces Tregs differentiation and function, maintains the immune homeostasis in the liver, and promotes hepatocyte proliferation and damage repair by secreting immunosuppressive factors, providing an important preliminary experimental basis for the development of new drugs based on these targets. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1A-J shows that the number of tolerogenic cDC1 was significantly reduced in AILF.

[0012] Figure 2 AH showed metabolic reprogramming of cDC1 in AILF.

[0013] Figure 3A-E shows that tolerogenic cDC1 improves AILF liver injury by inducing Tregs differentiation.

[0014] Figure 4 AM showed that P300 regulates the immune tolerance phenotype of cDC1 by mediating H3K27ac modification. DETAILED DESCRIPTION

[0015] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention. In addition, it should be understood that after reading the content taught by the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms fall within the scope limited by the appended claims of the application equally.

[0016] Example 1

[0017] This example is divided into three parts: ① The frequency, distribution changes and metabolic characteristics of tolerant cDC1 in the process of AILF; ② The regulatory effect of tolerant cDC1 on Tregs and its function in the process of AILF; ③ The specific mechanism and clinical pathological significance of P300-mediated H3K27ac modification regulating the immune tolerance phenotype of cDC1.

[0018] Part 1: Revealing the frequency, distribution changes and metabolic characteristics of tolerant cDC1 during the process of AILF.

[0019] Single cell RNA sequencing (scRNA-seq) was used to analyze the single cell profiles of liver transplant specimens from healthy liver donors and AILF patients. It was found that compared with healthy controls, the liver cDC1 of AILF patients was greatly reduced, and CD8 + T cell activation increased significantly, and was further verified by multiplexed immunohistochemistry (mIHC). A mouse AILF model was constructed, and mIHC results confirmed that cDC1 in the liver of AILF mice was significantly reduced. Flow cytometry results showed that the proportion of cDC1 decreased significantly with the extension of APAP intervention time and the increase of dose, suggesting that cDC1 may play an important role in the process of AILF (Figure 1A-J).

[0020] Single-cell gene set variation analysis (GSVA) ​​and metabolic flux analysis confirmed that the cDC1 subpopulation highly expressed IDO1 and that the activity of the tryptophan metabolic pathway was significantly upregulated. Intercellular communication analysis suggested that cDC1 and Tregs had close interactions ( Figure 2 AH).

[0021] Part 2: Clarify the regulatory effect of tolerant cDC1 on Tregs and its function in the process of AILF.

[0022] Mouse bone marrow-derived dendritic cells were isolated and induced to obtain tolerant cDC1 with high expression of IDO1, which was then adopted back into AILF model mice. It was found that adoptive back transfusion of tolerant cDC1 could significantly reduce APAP-induced liver injury and reduce inflammatory cell infiltration. At the same time, Ki67 staining showed a significant increase in liver regeneration. mIHC spatial in situ analysis found that adoptive back transfusion of tolerant cDC1 could induce a large increase in Tregs, and they formed clusters closely adjacent to each other in the damaged area, strongly suggesting that the two had a close interaction. (Figure 3A-E).

[0023] Part III: Elucidate the specific mechanism and clinical pathological significance of P300-mediated H3K27ac modification in regulating the immune tolerance phenotype of cDC1.

[0024] Single-cell SCENIC analysis showed that the key histone acetyltransferase P300 in epigenetic modification (rather than its homologous protein CBP) may be an important factor in maintaining the immune tolerance phenotype of cDC1. mIHC confirmed that P300, H3K27ac and IDO1 were co-localized in large quantities in the portal area of ​​human and mouse livers, and were relatively specifically expressed in cDC1, suggesting that IDO1 expression, which is potentially regulated by P300, may be one of the important mechanisms for maintaining the immune tolerance phenotype of cDC1. CUT&Tag sequencing was performed using H3K27ac antibodies to isolate BMDCs, and it was found that H3K27ac signals were enriched in the promoter region of the IDO1 gene. After intervening with BMDCs using a P300 inhibitor and performing conventional transcriptome sequencing, it was found that inhibiting P300 could significantly reduce the expression level of the IDO1 gene in BMDCs. P300 inhibitor was used to intervene in vitro induced tolerance BMDCs (tolDC) and H3K27ac CUT&Tag sequencing was performed. It was found that inhibition of P300 significantly reduced the binding of H3K27ac to the IDO1 promoter region. CHIP-qPCR verification reached the same conclusion. Conventional transcriptome sequencing also confirmed that P300 can significantly reduce the IDO1 gene expression level of tolDC ( Figure 4 AM).

Claims

1. Application of type I classical dendritic cells in the preparation of drugs for the treatment of drug-induced liver failure.

2. The use according to claim 1, characterized in that: The drug-induced liver failure includes acetaminophen-induced drug-induced liver failure.

3. The use according to claim 1, characterized in that: The medicine further comprises a pharmaceutically acceptable carrier and / or excipient.

4. The use according to claim 3, characterized in that: The pharmaceutically acceptable carrier and / or excipient includes at least one of a diluent, a binder, a surfactant, a wetting agent, an adsorption carrier, a lubricant, a filler, and a disintegrant.

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