Marker for evaluating gemcitabine chemosensitivity of intrahepatic cholangiocarcinoma and application of marker

A technique for intrahepatic cholangiocarcinoma and gemcitabine, which is applied in the field of medical biological detection

Pending Publication Date: 2020-07-28
中国人民解放军海军军医大学第三附属医院
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The research on the correlation between gemcitabine chemotherapy regimen and CXCR3 function level is still very limited, an...

Method used

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  • Marker for evaluating gemcitabine chemosensitivity of intrahepatic cholangiocarcinoma and application of marker
  • Marker for evaluating gemcitabine chemosensitivity of intrahepatic cholangiocarcinoma and application of marker
  • Marker for evaluating gemcitabine chemosensitivity of intrahepatic cholangiocarcinoma and application of marker

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1: Peripheral blood sample processing

[0049] 1. Sample processing

[0050] Peripheral blood was obtained from 20 ICC patients (8 of whom were sensitive to chemotherapy and 12 of whom were insensitive to chemotherapy) in the Eastern Hepatobiliary Surgery Hospital, who were treated with gemcitabine chemotherapy.

[0051] The overall workflow for isolating leukocytes from peripheral blood for CyTOF analysis is as follows: figure 1 shown. 2 mL of peripheral blood was collected at three time points before, during and after chemotherapy, and a total of 49 samples were obtained for this experiment. All samples were anonymously coded in accordance with local ethical guidelines.

[0052] 2. Isolation of white blood cells from peripheral blood

[0053] Leukocytes were separated from the fresh peripheral blood described above to obtain 20 groups of peripheral blood mononuclear cells (PBMC).

[0054] The specific steps are:

[0055] Dilute whole blood samples 1:1...

Embodiment 2

[0089] Example 2 Spatial heterogeneity analysis of peripheral blood immune microenvironment in intrahepatic cholangiocarcinoma

[0090] This example collects high-dimensional single-cell proteomic profiles from nearly 20,000,000 leukocytes (an average of approximately 450,000 cells per sample). The distribution of the immune lineage is visualized by tSNE as shown in figure 2 Shown: There are significant differences in the number of T cells, B cells, NK cells, monocytes, and mDCs in the peripheral blood of R (chemotherapy sensitive) and NR (chemotherapy insensitive) patients with ICC. This indicates that there is great heterogeneity in the peripheral blood immune microenvironment among different ICC patients.

Embodiment 3

[0091] Example 3 Individual differences of T cell clusters and CXCR3 before chemotherapy + Distribution characteristics of T cells

[0092] To investigate T cell subset composition in R versus NR patients, we visualized and reanalyzed T cell subsets. By applying the FlowSOM algorithm, T cells can be divided into 29 clusters (cluster) (including all T cells from 20 patients at three time points for analysis) ( image 3 and Figure 4 ). There were significant differences in T cell clusters among different individuals (clusters 1, 16, and 22 were more significant), and R and NR patients could not be well clustered and separated.

[0093] In order to further search for commonalities and differences between R and NR, we clustered the T cells of the samples at three time points with the above-mentioned significantly different T cell clusters (clusters 1, 16, 22) ( Figure 5 ). Before gemcitabine treatment, clusters 1, 16, and 22 of the two groups of patients with different chem...

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Abstract

The invention discloses a marker for evaluating the gemcitabine chemosensitivity and/or prognosis of intrahepatic cholangiocarcinoma and application of the marker. The marker for evaluating the gemcitabine chemosensitivity and/or prognosis of intrahepatic cholangiocarcinoma is CXCR3 expressed on the surfaces of peripheral blood leukocytes or immune cells in cholangiocarcinoma tissues of an intrahepatic cholangiocarcinoma patient. A kit for evaluating the gemcitabine chemosensitivity and/or prognosis of intrahepatic cholangiocarcinoma comprises a reagent related to detection of the expression quantity of the CXCR3 protein. A method for evaluating the gemcitabine chemosensitivity and/or prognosis of intrahepatic cholangiocarcinoma comprises the following steps: (1) detecting the expression quantity of CXCR3 on the surfaces of peripheral blood leukocytes or immune cells in cholangiocarcinoma tissues of an intrahepatic cholangiocarcinoma patient; and (2) comparing the expression quantity with a preset CXCR3 protein expression quantity threshold value, evaluating that the patient is sensitive to gemcitabine chemotherapy and/or good in prognosis of gemcitabine chemotherapy if the CXCR3 protein expression quantity is higher than the threshold value, and otherwise, evaluating that the patient is insensitive to gemcitabine chemotherapy and/or poor in prognosis of gemcitabine chemotherapy.

Description

technical field [0001] The invention belongs to the technical field of medical biological detection, and relates to a marker, a kit and a method for evaluating gemcitabine chemotherapy sensitivity and prognosis of intrahepatic cholangiocarcinoma. Background technique [0002] Intrahepatic cholangiocarcinoma (ICC), which occurs in the proximal secondary bile ducts, is the second most common primary liver cancer, and its incidence is steadily increasing. ICC is derived from intrahepatic bile duct epithelial cells, and surgical resection is the only effective treatment. As a highly malignant disease, only patients who undergo complete R0 surgical resection can have a better survival rate. Studies have confirmed that radiotherapy and adjuvant chemotherapy are beneficial to the survival of advanced or aggressive cholangiocarcinoma. The preferred treatment for unresectable and metastatic ICC is the combination of gemcitabine and cisplatin. However, ICC patients respond differentl...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/577
CPCG01N33/57407G01N33/57492G01N33/577G01N2333/715
Inventor 王红阳陈磊杨应成吴彤
Owner 中国人民解放军海军军医大学第三附属医院
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