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Bladder cancer depleted NKT cell subset, characteristic gene and application thereof

A technology of NKT cells and genes, applied in the field of tumor molecular biology, can solve the problems of TIL types and inhibition pathways that are not fully understood

Pending Publication Date: 2020-10-02
BEIJING UNIV OF CHEM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the mechanism of interaction between the immune system and cancer tissue plays an important role in the development and improvement of immunotherapy, as well as in the detection of tumor development and progression, the types of TILs and their inhibitory pathways have not been fully understood

Method used

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  • Bladder cancer depleted NKT cell subset, characteristic gene and application thereof
  • Bladder cancer depleted NKT cell subset, characteristic gene and application thereof
  • Bladder cancer depleted NKT cell subset, characteristic gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Single Cell Transcriptome Sequencing

[0043] 1. Clinical sample collection

[0044] In this study, peripheral blood, bladder cancer tissues and adjacent normal tissues of 3 patients with bladder urothelial carcinoma were selected, which were collected and provided by Xuanwu Hospital of Capital Medical University. All patients provided written informed consent. Surgically resected bladder cancer tissues, adjacent normal tissues and peripheral blood were collected from patients. All patients were required to have no history of autoimmune diseases or other cancers, bladder cancer who had not received neoadjuvant chemotherapy or other anti-tumor treatments before surgery, and the tumor diameter was greater than 1cm. For sample collection, the fresh sample should be transported back to the laboratory at 4°C immediately after the operation, and the cell separation experiment should be carried out within 2 hours to ensure the maximum activity of the cells.

[004...

Embodiment 2

[0068] Example 2 Gene expression analysis of different cell subpopulations

[0069] Through single-cell transcriptome sequencing, according to Cellranger data statistics, Seurat standardization, dimensionality reduction, clustering, etc., the sequencing results were analyzed, and clustering was performed according to the gene expression between cells. Cluster analysis was used to identify cell subtypes. According to the clustering results, the t-SNE dimensionality reduction algorithm is used to display the distribution of cells in two-dimensional space. The specific analysis method is as follows:

[0070] (1) Data standardization and noise reduction

[0071] Seurat uses the global normalization method "LogNormalize" to standardize the gene expression matrix by default. For each cell, the expression of each gene is divided by the overall expression of the cell, that is, converted to relative abundance, and then multiplied by Normalization factor (default 10000), and then perf...

Embodiment 3

[0094] Example 3CD8 + Pathway enrichment analysis of differentially expressed genes in exhausted NKT cells

[0095] Gene pathway enrichment analysis is the process of analyzing the metabolic pathways and functions of genes in cells. In the present invention, Matescape is used to perform pathway enrichment analysis on differentially expressed genes. Metascape is a portal website that provides gene annotation and analysis resources. The website is http: / / metascape.org. It integrates more than 40 biological information databases, such as GO / KEGG terms, canonical pathways, hall mark gene sets, UniProt and DrugBank, etc. Commonly used databases are included. We can conveniently use this website for functional enrichment analysis of biological pathways and protein interaction network structure analysis. We submit the list of differential genes of the cluster to be analyzed on the homepage of the website, select the species information as H.sapiens, and then click the analysis opti...

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Abstract

The invention discloses a bladder cancer depleted NKT cell subset, a characteristic gene and application thereof. The bladder cancer depleted NKT cell subset is characterized in that a single cell transcriptome sequencing analysis technology is utilized to perform single cell transcriptome sequencing on NKT cells from peripheral blood, normal bladder tissues and bladder cancer tissues of an infiltrative bladder cancer patient, a bladder cancer specific NKT cell subset, namely CD8<+> depleted NKT cells, is identified, and characteristic genes expressed by the CD8<+> depleted NKT cells, namely genes CSF2, ATF3, BAG3, NR4A1 and NR4A2, are determined. Moreover, the characteristic genes can be used for identifying or identifying the CD8<+> depleted NKT cells from the bladder cancer, and furtherused for auxiliary diagnosis, prognosis and immunotherapy of the bladder cancer, and provide a related detection reagent or kit.

Description

technical field [0001] The invention relates to the fields of tumor molecular biology and biotechnology, in particular to the identification of cell subgroups, especially the identification of NKT cell subgroups derived from bladder cancer tissue. Background technique [0002] Bladder cancer is one of the most common malignant tumors of the urinary system. In 2018, CA magazine reported that there were about 549,000 new cases and 200,000 deaths from bladder cancer worldwide, and men accounted for about 75% of the overall disease burden. Bladder cancer is the fourth most common cancer in the United States, affecting four times as many men as women. In my country, the incidence rate of bladder cancer is 6.69 / 100,000, and the incidence rate shows a trend of increasing with time, which greatly threatens the health of our people, especially male patients. Bladder cancer will have great differences due to differences in gender, race and region. It can occur in all age groups, and...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886G01N33/68G01N33/574C12N15/11C12N5/0783
CPCC12Q1/6886G01N33/68G01N33/57407G01N33/57484C12N5/0646C12Q2600/118C12Q2600/158G01N2800/52G01N2800/60
Inventor 喻长远杨昭王璐瑶白素杭沈宗毅
Owner BEIJING UNIV OF CHEM TECH
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