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Analysis of response to therapeutics in cancer

A technology for cancer treatment and response, applied in analytical materials, electrophoresis analysis, biomaterial analysis, etc., can solve problems such as complex signal networks

Inactive Publication Date: 2020-12-11
THE BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIOR UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

More than 2,000 human genes are estimated to encode kinases, and each kinase has the potential to act on multiple targets, resulting in a complex signaling network

Method used

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  • Analysis of response to therapeutics in cancer
  • Analysis of response to therapeutics in cancer
  • Analysis of response to therapeutics in cancer

Examples

Experimental program
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Effect test

example 1

[0104] Effect of specimen storage time and temperature on ERK phosphorylation

[0105] Effect of specimen storage time on ERK phosphorylation. An important issue for the stability of ERK phosphorylation in clinical fine needle aspiration (FNA) specimens is the time interval between FNA collection and FNA snap freezing in the laboratory. We refer to this interval as the FNA storage time. Additionally, the temperature at which FNA is stored may affect ERK phosphorylation. To address these questions, we performed a benchmark test to validate the stability of ERK phosphorylation in FNA from RCC patients (see below). The relative abundance of phosphorylated and non-phosphorylated ERK isoforms was remarkably stable, suggesting that our measurements were not significantly altered by FNA treatment, even when stored for up to 2 days.

[0106] For the three FNAs from different patients, one part of the FNAs was processed and snap-frozen on the same day of tissue collection, and the o...

example 2

[0115] Such as image 3 As shown, fine-needle aspirates from two regions of the same human RCC tumor (T1 and T2) and adjacent renal tissue (N) were obtained from four patients with renal cancer, and glutaminase between samples was provided. level distribution. Charge-separation NIA was used to measure the levels of the KGA and GAC isoforms of glutaminase 1 (GLS1 ) in these samples using anti-GLS1 antibodies. This demonstrates that NIA analysis of protein isoforms is involved in metabolism.

[0116] Such as Figure 4 As shown, NIA can measure the distribution of isoforms of the antioxidant protein peroxidase 6 (PRDX6) from frozen samples. Cells from each patient were analyzed with an anti-PRDX6 antibody (Abcam, Cat# 73350). Patients had EGFR mutation (MUT), wild type (WT) or unknown EGFR status (A, B, C, D).

[0117] Figures 5A-5I The isoform distribution of human carbonic anhydrase 9, human α-tublin (tubulin) is shown in a series of NIA assays. Under some conditions, t...

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Abstract

A nanoimmunoassay (NIA) is applied to quantify analytes, including without limitation proteins and isoforms of proteins involved in oncogenic or metabolic signaling pathways, in a small amount of lysate from a tissue sample. Samples of interest for NIA include without limitation blood or solid tumor microbiopsy samples, such as fine needle aspirate (FNA) or circulating tumor cells. Samples may betaken at a single timepoint, or may be taken at multiple timepoints. Samples may be as small as 100,000 cells, as small as 5000 cells, as small as 1000 cells, as small as 100 cells, as small as 50 cells, as small as 25 cells or less. The NIA detection method combines size separation of proteins or isoelectric protein focusing and antibody detection in a microfluidic system.

Description

[0001] cross reference [0002] This patent application claims priority over the filing dates of U.S. Provisional Patent Application No. 62 / 740,013, filed October 2, 2018, and U.S. Provisional Patent Application No. 62 / 644,303, filed March 16, 2018 Rights, the contents of the above patents are incorporated herein by reference. [0003] Federally Funded Research and Development [0004] The patent of this invention has obtained the support of the government, among which, the funding project of the National Institutes of Health of the United States is CA196585. The government has certain rights in this invention. Background technique [0005] Transformation and growth of tumor cells is a complex process that can vary even within specific tissue types. Assays that can define the phenotype of tumor cells can be used to determine appropriate therapy and thus have clinical implications. In addition, understanding the mechanisms associated with onset of therapy allows for the det...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B82Y15/00G01N33/00G01N33/50G01N33/574
CPCG01N33/5011G01N2550/00G01N2800/52G01N33/57484B82Y5/00G01N33/5748B82Y15/00C12Q1/025
Inventor 阿文·古乌爱丽丝·凡迪恩·W·费尔舍
Owner THE BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIOR UNIV
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