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Method and detection kit for judging whether a solid tumor is suitable for immunotherapy

A technology for immunotherapy and detection reagents, applied in biochemical equipment and methods, microbial determination/inspection, measurement devices, etc., can solve problems such as immune system identification and tumor-killing ability evaluation, achieve objective response rate improvement, and broad application Foreground effect

Active Publication Date: 2022-04-29
LEPU MEDICAL TECH (BEIJING) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The biomarkers discovered so far are all focused on local events, and have not evaluated the ability of the immune system to recognize and kill tumors globally

Method used

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  • Method and detection kit for judging whether a solid tumor is suitable for immunotherapy
  • Method and detection kit for judging whether a solid tumor is suitable for immunotherapy
  • Method and detection kit for judging whether a solid tumor is suitable for immunotherapy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Example 1 Detection of mutation load

[0072] From April 2016 to November 2017, FFPE (formaldehyde-fixed paraffin-embedded) samples and peripheral blood of tumor tissues from 80 patients with solid tumors were collected. The peripheral blood cfDNA (circulating free DNA) and peripheral blood genomic DNA of these 80 patients with solid tumors were extracted, and the sequencing library was constructed, hybridized and captured, and sequenced on a computer, and bioinformatics analysis was performed on the obtained sequencing data. The process can be found in figure 1 . The tumor mutation burden of peripheral blood CtDNA was calculated according to the following formula 4.

[0073] Mutation load = total number of variations in the sample assay region / size of the target region (Formula 4)

Embodiment 2

[0074] Example 2 Detection of HLA heterozygous deletion

[0075] The tumor tissue and paired peripheral blood genomic DNA of 80 solid tumor patients described in Example 1 were extracted, and seven polymorphic STR sites located in the genomic region where the HLA gene was located were selected, and the seven STRs were detected by fluorescent PCR-capillary electrophoresis The homozygous status of the loci (7 loci are D6S2852, D6S2872, D6S248, D6S1022, D6S265, D6S273, D6S1666), and the heterozygous deletion ratio of each locus is calculated according to the following formula 5. In formula 5, allele 1 is the allele with lower peak height. If the heterozygous deletion ratio is higher than 0.4, it is considered that the heterozygous deletion occurs at this site. Compared with the peripheral blood genomic DNA, if there is a heterozygous deletion at one site, then The HLA status of this sample is heterozygous deletion.

[0076] Heterozygous deletion ratio=1-Ht value=1-[peak height o...

Embodiment 3

[0079] Example 3 Calculate the mRNA expression scores of 18 genes related to T cell inflammation

[0080] RNA was extracted from the tumor tissues of 80 patients with solid tumors in Example 1, and 18 genes related to T cell inflammation (see Table 1) were selected, reverse-transcribed into cDNA, and a fixed amount of external standard was added to the cDNA sample.

[0081] The Ct value of 18 genes related to T cell inflammation, 1 internal reference gene β-actin, and 1 external standard was detected by Q-RT-PCR method (based on taqman probe), and the expression score of each gene was calculated. Then calculate the average value, which is the mRNA expression score of the 18 T cell inflammation-related genes. The calculation method is described below taking the CXCR6 gene as an example: Absolute expression level of CXCR6=absolute copy number of external standard product*2 -ΔΔCt , while ΔΔCt=[Ct(CXCR6)-Ct(internal reference gene)]-[Ct(external standard)-Ct(reference gene)], the...

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Abstract

The invention provides a detection kit for judging whether a solid tumor in a patient is suitable for immunotherapy, which includes detection reagents for detecting the following indicators: tumor mutation load of peripheral blood circulating tumor DNA, HLA typing, HLA Heterozygous deletion status, percentage of tumor cells positive for PD‑L1 membrane expression, CD8 + Infiltration levels of immune cells, FOXP3 + Infiltration level of immune cells, mRNA expression score of T cell inflammation-related genes, CD14 in peripheral blood mononuclear cells + CD16 ‑ HLA‑DR + Percentage of monocytes, and microsatellite instability status. Correspondingly, the present invention also provides a method for judging whether a solid tumor in a patient is suitable for immunotherapy. After the screening of solid tumor patients by using the kit and method of the present invention, the objective response rate is significantly improved, and has broad application prospects in clinical screening of solid tumor patients suitable for immunotherapy.

Description

technical field [0001] The invention relates to a method for judging whether a solid tumor is suitable for immunotherapy, especially a method for judging whether a solid tumor is suitable for immunotherapy by using multi-index joint detection and a logistic regression model. The invention also relates to detection kits that can be used in this method. Background technique [0002] Tumor immunotherapy is the most exciting development in cancer treatment in recent years, especially PD-1 / PD-L1 immunotherapy has become a hot topic today. Although PD-1 / PD-L1 blockers have achieved good curative effect and high tolerance in clinical practice, there will be significant individual differences in the clinical application of immune checkpoint drugs, and different patients will not respond to the same drug. different drug sensitivities. Except in advanced Hodgkin's lymphoma, the objective response rate of PD-1 blockade was 60%. In tumors, the objective response rate did not exceed 3...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/02C12Q1/6886
CPCG01N33/5005C12Q1/6886C12Q2600/106C12Q2600/158
Inventor 韩玉卿徐芹李俊辉张江生王凯琳蒲珏
Owner LEPU MEDICAL TECH (BEIJING) CO LTD
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