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Salivary Biomarkers for Gastric Cancer Detection

a biomarker and gastric cancer technology, applied in the field of salivary biomarkers for gastric cancer detection, can solve the problems of high risk of gastric cancer in the first-generation of gastric cancer patients, and high incidence of gastric cancer in japan and korea, so as to improve the prognosis of patients and greatly enhance the survival of cancer patients

Inactive Publication Date: 2018-07-05
RGT UNIV OF CALIFORNIA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The patent text discusses the importance of early diagnosis and treatment of gastric cancer to improve survival outcomes. The text describes a method for assessing the efficacy of therapy on a subject by analyzing specific cancer markers in saliva samples. The method involves collecting a first saliva sample before initiating therapy and analyzing it to detect specific cancer markers. After implementing therapy, a second saliva sample is collected and analyzed to compare the cancer marker profile before and after treatment. This method can help track the response of therapy and provide valuable information for the treatment of gastric cancer.

Problems solved by technology

Gastric cancer is a highly aggressive and lethal malignancy.
Asian countries, such as Japan and Korea, have particularly high incidents of gastric cancer.
First-generation offspring of gastric cancer patients and people with blood type A are at increased risk of gastric cancer.
In addition to heredity and diet, infection with Helicobacter pylori (Campylobacter pyloridis) is a risk factor for developing gastric cancer.
Gastric cancer often goes undiagnosed until detected in an advanced state thereby limiting ameliorative treatment options.
Currently, a diagnosis of gastric cancer is made by gastrointestinal X-ray examination and endoscopic examination both of which have particular limitations.
Both of these methods require highly trained specialists and are impractical as a general screening method.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

Salivary Transcriptomic Profiling and Analysis

[0142]After collection, saliva was centrifuged at 2600 g at 4° C. for 15 minutes. Saliva supernatant was separated from the pellet. Total RNA from the saliva supernatant samples of 63 gastric cancer patients and 31 healthy controls were extracted using an RNA extraction kit (Qiagen RNeasy Mini Kit from Qiagen). DNase treatment (TURBO™ DNase, Ambion) was used to remove contaminating DNA. 90 μL of extracted total RNA (out of 100 μL) was concentrated to 10 μL and then linearly amplified using the RiboAmp RNA Amplification kit (Molecular Devices, Sunnyvale, Calif.). After purification, cDNA was in vitro transcribed and biotinylated using GeneChip Expression 3′-Amplification Reagents for in vitro transcription labeling (Affymetrix, Santa Clara, Calif.). Equal amounts of labeled RNA (˜20 μg) were subsequently fragmented and sent to the UCLA microarray core facility for chip hybridization and scanning. The Affymetrix Human Genome U133 Plus 2.0 ...

example 2

Salivary Proteomic Profiling and Analysis

[0148]After collection, saliva was centrifuged at 2600 g at 4° C. for 15 minutes. Saliva supernatant was separated from the pellet and stored at −80° C. until further analysis. By taking equal amount of protein from each individual sample (20 gastric cancer samples and 20 healthy control samples), proteins from every four samples were pooled into one sample in the cancer group and healthy control group, respectively. These result in 5 pooled cancer samples and 5 pooled healthy control samples. All the 10 pooled samples were subjected to amylase removal by using starch column. Two global internal standard (GIS) pooled saliva samples were made from all the 10 pooled samples for the comparison between two TMT-6plex experiments.

[0149]For one TMT-6plex experiment, the 100 μg proteins in each saliva sample were dissolved in 45 μL of 200 mM TEAB, adjust the sample to a final volume of 100 uL with ultrapure water. With adding 5 μL 200 mM TCEP, the re...

example 3

Saliva Microbial Biomarkers

[0155]After collection, saliva was centrifuged at 2600 g at 4° C. for 15 minutes. The pellet was separated from the saliva supernatant and DNA from the saliva pellet was extracted using UltraClean Microbial DNA Isolation Kit (MO BIO Laboratories Inc.). The protocol followed the product manual. PCR amplification using 16S universal primers was performed in Forsyth Institute, followed by hybridization on the HOMIM microarray. Selection of bacteria candidates: based on the fold change and statistical analysis (Wilcoxon signed-rank test), analysis from the UCLA Wong lab and Forsyth Institute were combined. Twenty-eight candidates were selected for differentiation between disease and control.

[0156]Confirmation by real time PCR: quantities of bacteria species in the original DNA samples were determined using real time PCR. Specific primers were designed for all species and real-time PCR are performed to check the bacterial quantity between cancer and controls. T...

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Abstract

Disclosed herein are biomarkers related to gastric cancer. The presently identified salivary biomarkers create the basis for a gastric cancer detection bioassay with sensitivity and specificity. Means and methods for evaluating the data generated using multiple biomarkers in order to validate findings and further use of the multiplexed gastric cancer assay in clinical, diagnostic and therapeutic uses is also included.

Description

CROSS-REFERENCES TO RELATED APPLICATIONS[0001]The present application claims priority to U.S. Ser. No. 61 / 532,210, filed Sep. 8, 2011, herein incorporated by reference in its entirety.STATEMENT OF GOVERNMENT RIGHTS[0002]This invention was made with U.S. Government support under Grant No. DE016275 awarded by the National Institutes of Health. The U.S. Government has certain rights in the invention.BACKGROUND[0003]Gastric cancer is a highly aggressive and lethal malignancy. On a global basis, gastric cancer is the second leading cancer cause of death. Every year nearly 700,000 people die from the disease. Asian countries, such as Japan and Korea, have particularly high incidents of gastric cancer. For example, it is estimated that gastric cancer causes more than 16% of the male deaths in Korea and Japan.[0004]Both heredity and environmental factors play roles in the development of gastric cancer. First-generation offspring of gastric cancer patients and people with blood type A are at...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G01N33/574C12Q1/6886
CPCG01N2800/52C12Q1/6886C12Q2600/158G01N33/57446
Inventor WONG, DAVID T.
Owner RGT UNIV OF CALIFORNIA
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