Search

CN-122012763-A - Application of lncRNA in diagnosis and treatment of mycobacterium tuberculosis infection

CN122012763ACN 122012763 ACN122012763 ACN 122012763ACN-122012763-A

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to application of lncRNA in tuberculosis infection. In the invention, a plurality of lncRNAs related to M.tb infection are identified by taking the M.tb infected THP-1 cells as a model, and LNCRNA ENST00000608721 is proved to be highly expressed in tuberculosis infection. By detecting the expression level, tuberculosis infection state and non-infection state can be effectively distinguished. Further studies found that CFU in cells silenced LNCRNA ENST00000608721 was significantly lower than that in control cells, indicating that knockdown LNCRNA ENST00000608721 enhanced the ability of macrophages to clear m.tb, exerting bacteriostatic effects. It is shown that ENST00000608721 can be used by M.tb manipulation as a negative regulatory factor of host anti-tuberculosis infection immune response to help M.tb realize immune escape.

Inventors

  • PAN LIPING
  • ZHAO YUEHAN
  • DONG JING
  • Shang Xuetian
  • WANG XINYU
  • JI XIUXIU
  • ZHANG LANYUE
  • ZHU CHUANZHI
  • ZHANG ZONGDE

Assignees

  • 首都医科大学附属北京胸科医院
  • 北京市结核病胸部肿瘤研究所

Dates

Publication Date
20260512
Application Date
20260312

Claims (10)

  1. 1. A biomarker for diagnosing tuberculosis infection, which is characterized in that the biomarker is LNCRNA ENST00000608721, and the nucleotide sequence of the biomarker is shown as SEQ ID NO. 1.
  2. 2. The application of a reagent for detecting LNCRNA ENST00000608721 relative expression quantity in preparing a reagent for diagnosing tuberculosis infection is characterized in that the LNCRNA ENST00000608721 nucleotide sequence is shown as SEQ ID NO. 1.
  3. 3. A kit for diagnosing tuberculosis infection, comprising the reagent of claim 2.
  4. 4. The kit of claim 3, wherein the kit is selected from one or more of a nucleic acid detection kit, an immunofluorescence kit, a gene chip detection kit, a molecular hybridization kit, and/or an in situ hybridization staining kit.
  5. 5. The application of a reagent for inhibiting LNCRNA ENST00000608721 expression in preparing a medicament for treating tuberculosis infection is characterized in that the reagent is an siRNA reagent, the siRNA reagent is a mixed library composed of three pairs of siRNAs and 3 ASOs, the sequence of the siRNA is shown as SEQ ID NO.2-SEQ ID NO.4, and the sequence of the ASOs is shown as SEQ ID NO.5-SEQ ID NO. 7.
  6. 6. The use according to claim 5, wherein one or more pharmaceutically acceptable carriers are further added to the medicament.
  7. 7. The use of claim 5, wherein the pharmaceutical dosage form comprises, but is not limited to, one or more of a tablet, capsule, aerosol, pill, powder, solution, suspension, emulsion, granule, liposome, transdermal, and/or suppository.
  8. 8. The use according to claim 5, wherein the medicament is used alone or in combination with other medicaments.
  9. 9. A pharmaceutical composition for treating tuberculosis infection is characterized in that an active ingredient of the pharmaceutical composition comprises an agent for inhibiting LNCRNA ENST00000608721 expression, wherein the agent is an siRNA agent, the siRNA agent is a mixed library composed of three pairs of siRNAs and 3 ASOs, the sequence of the siRNA is shown as SEQ ID NO.2-SEQ ID NO.4, and the sequence of the ASOs is shown as SEQ ID NO.5-SEQ ID NO. 7.
  10. 10. An animal model of tuberculosis infection, wherein the animal model is high expression LNCRNA ENST00000608721 of the animal.

Description

Application of lncRNA in diagnosis and treatment of mycobacterium tuberculosis infection Technical Field The invention belongs to the technical field of biological medicines, and particularly relates to application of lncRNA in tuberculosis infection. Background Mycobacterium tuberculosis (Mycobacterium tuberculosis, M.tb) is an intracellular pathogen that severely threatens human health, macrophages serve as the primary target cells for M.tb infection, and play a central role in the host's resistance to M.tb infection. It is known in the prior art that m.tb is engulfed by host macrophages, and then the host molecule can be used by a variety of mechanisms to counteract or induce the host molecule to evade the clearance of the natural immune defense mechanism, so as to achieve survival and proliferation in the cell, thereby establishing persistent infection （Chai Q, Wang L, Liu CH, Ge B. New insights into the evasion of host innate immunity by Mycobacterium tuberculosis. Cell Mol Immunol. 2020;17(9):901-913;Kaufmann SHE, Dorhoi A, Hotchkiss RS, Bartenschlager R. Host-directed therapies for bacterial and viral infections. Nat Rev Drug Discov. 2018;17(1):35-56.）. Tuberculosis (TB), which is a chronic infectious disease caused by m.tb infection, and also a disease in which single bacterial infection causes the most death in addition to the novel coronavirus. Improving the cure rate of tuberculosis, especially drug-resistant tuberculosis, is one of the key measures for reducing morbidity and mortality, so that deep and systematic research on tuberculosis infection and pathogenic mechanism is urgently needed, new drug targets are found, and development of novel anti-tuberculosis drugs is promoted. Macrophages, the main parasitic cells of m.tb in the host, are the first line （WANG Z, XU H, WEI Z, et al. The role of non-coding RNA on macrophage modification in tuberculosis infection. Microb Pathog. 2020;149: 104592.）, of defense of the host against infection, which is both the key executor of the innate immune response and the initiator of adaptive immunity. The prior theory holds that when m.tb invades the host's lungs, alveolar macrophages capture bacteria by phagocytosis and initiate a range of killing mechanisms including acidification of the phagosome and its fusion with lysosomes, induction of apoptosis, release of bactericidal substances such as reactive oxygen species and nitric oxide, and triggering inflammatory responses and release of type I interferons by activating the inflammatory small body and cGAS pathways. However, m.tb also alters the host immune response by stress or induction or the like in a different way or initiates host metabolic reprogramming, utilizing host molecules to accomplish their survival and proliferation within the cell. Identification of key host molecules during m.tb infection of macrophages and analysis of their function is the key and basis for the development of host-directed anti-tuberculosis therapeutic drugs. At least 95% of human genome transcripts are non-coding RNAs (ncrnas), of which about 80% are Long non-coding RNAs (lncRNAs) greater than 200 nucleotides in length, which do not possess protein coding ability, but enable fine gene expression regulation （Quinn JJ, Chang HY. Unique features of long non-coding RNA biogenesis and function [J]. Nat Rev Genet. 2016; 17 (1): 47-62.）. at multiple levels of transcription, post-transcription, translation, etc. by direct binding to DNA, RNA or protein, in the tuberculosis field, existing research on lncRNAs is mostly limited to screening levels of molecular markers, while data on functional research is less. Most current studies have found that host lncRNA molecules may be involved in macrophage-mediated anti-tuberculosis infection with the identity of positive regulatory factors. Disclosure of Invention According to the invention, a plurality of lncRNAs related to M.tb infection are identified by taking the THP-1 cells infected by the M.tb as a model, and the high expression characteristics of LNCRNA ENST00000608721 are confirmed, and further research shows that after silencing the molecules, the capacity of eliminating the M.tb of macrophages can be reduced, so that the molecules are controlled by the M.tb and serve as negative regulatory factors of host anti-tuberculosis infection immune response, and the M.tb is helped to realize immune escape. Based on this, the present invention has been completed. In a first aspect, the invention provides a biomarker for diagnosing tuberculosis infection, wherein the biomarker is LNCRNA ENST00000608721, and the nucleotide sequence of the biomarker is shown as SEQ ID NO. 1. Further, the Mycobacterium tuberculosis infection includes primary infection, secondary infection and extrapulmonary infection. In a second aspect, the invention provides an application of a reagent for detecting LNCRNA ENST00000608721 relative expression level in preparing a reagent for diagnosing tuberculosis infecti