Cancer Associated Pericyte (CAP) subset and preparation method and application thereof

A cell subset and perivascular technology, applied in the biological field, can solve the problems of increasing tumor metastasis, sensitizing small molecule targeted drugs and immunotherapy, and achieve the effect of promoting invasion and increasing inhibitory effect.

Pending Publication Date: 2021-12-31
THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, vascular pericytes are located at the junction of endothelial cells, immune cells and tumor cells, and play an important role as a portal in the metastatic microenvironment. The core of anti-angiogenesis therapy for tumors is to inhibit angiogenesis, which aims to deprive tumors of oxygen and nutrients. However, more and more evidence shows that anti-vascular therapy can increase the risk of tumor metastasis while inhibiting tumor growth, and the mechanism of this classic paradox is closely related to the concept of normalization of tumor blood vessels. It is to reshape the tumor vascular structure, improve the vascular delivery route of clinical drugs, and theoretically can significantly increase the sensitivity of small molecule targeted drugs and immunotherapy

Method used

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  • Cancer Associated Pericyte (CAP) subset and preparation method and application thereof
  • Cancer Associated Pericyte (CAP) subset and preparation method and application thereof
  • Cancer Associated Pericyte (CAP) subset and preparation method and application thereof

Examples

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Effect test

Embodiment 1

[0086] Example 1 Single-cell transcription analysis of tumor microenvironment in invasive lung adenocarcinoma and identification of CAP characteristic phenotype proteins

[0087]Using the single-cell transcriptome sequencing technology of 10X Genomics, a total of twelve samples (nearly 70,000 cells) of the primary tumor, metastatic positive lymph nodes, and negative lymph nodes of four patients with invasive lung adenocarcinoma with a mutation background were subjected to single-cell transcriptome sequencing ( figure 1 ); and through quality control, normalization correction, deletion integration and visualization preprocessing, the resulting sequences were annotated with whole-cell clusters using the SingleR embedded reference dataset ( figure 2 , image 3 , Figure 4 ), a panorama showing the gene expression profile of the tumor microenvironment of invasive lung adenocarcinoma, in which feature selection is performed on the stromal cells and pericytes (Pericyte, PC) in the...

Embodiment 2

[0089] Example 2 Separation and purification of lung cancer tumor-associated pericyte subpopulation

[0090] Lung cancer pericytes were separated by a self-built enzyme digestion system. The specific steps were as follows: clinically obtained fresh lung cancer tissue blocks after surgical resection, rinsed with PBS three times, placed in a centrifuge tube containing 50 mL of RPMI1640 cell culture medium, and sealed with sealing glue. Immediately store at 4°C and transport to the laboratory; take out the lung cancer tissue block in a biological safety cabinet and place it in a 10cm culture dish, use a scalpel to cut the lung cancer tissue into pieces smaller than 0.5cm, and remove the fat around the lung cancer tissue block and fibrous connective tissue; add 2mL containing 4mg / mL Dispase II (Dispase II), 0.6mg / mL Type I collagenase (Collagenase I), 0.6mg / mL Type IV collagenase (Collagenase IV) and 0.3 mg / mL DNase I (DNase I) RPMI-1640 medium; put the culture dish containing lun...

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Abstract

The invention belongs to the technical field of biology, and discloses a Cancer Associated Pericyte (CAP) subset and a preparation method and application thereof. The CAP subset expresses GJB2. According to the invention, a high-dimensional single-cell sequencing technology is utilized to perform single-cell transcriptome analysis and downstream cell functional verification on primary lesions, metastatic positive lymph nodes and non-metastatic lymph nodes of an infiltrating lung adenocarcinoma patient; the invention discloses for the first time that an independent CAP subset exists in an infiltrating lung adenocarcinoma tumor microenvironment; pDGFRb+GJB2+CSPG4- is specifically expressed as an identity tag, then the subset is successfully sorted, amplified and breed-preserved through characteristic antibody flow purification and immunofluorescence identification, and by further cell function verification, the subset can obviously promote the invasion ability of lung cancer cells, has the juvenile and malformed vascularization ability, and is an ideal cell tool for studying a lung cancer tumor vascular abnormality mechanism and a targeted vascular normalization drug in the field.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a tumor-associated vascular pericyte subgroup, a preparation method and application thereof. Background technique [0002] In recent years, the research hotspot in the field of tumors is the interaction between various cells in the tumor microenvironment and tumor cells, and to explore the functional mechanism of regulating tumor development and metastasis. With the in-depth application of single-cell omics, researchers have been able to Explore the impact of tumor microenvironment (Tumor Microenvironment, TME) heterogeneity on tumor progression in a higher dimension, in which the fibrous skeleton and heterogeneous vascular structure formed by stromal cells provide a substantial basis for tumor survival, invasion and immune escape. [0003] Vascular pericytes (Pericytes, PCs) are ubiquitous in the tumor microenvironment. The latest research has found that pericytes can ac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071G01N33/574C12Q1/02A61K45/00A61P35/00
CPCC12N5/069G01N33/57484G01N33/5011A61K45/00A61P35/00C12N2509/00C12N2509/10
Inventor 劳深何建行梁文华曾健明熊亚明陈子盛
Owner THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
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