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Method for predicting biological systems responses in hepatocytes

A hepatocyte and biological technology, applied in the field of predicting the response of biological systems in hepatocytes, can solve the problem of missing multiple functions of hepatocytes

Inactive Publication Date: 2010-07-21
CELLUMEN INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, HepG2 cells lack many functions of differentiated hepatocytes

Method used

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  • Method for predicting biological systems responses in hepatocytes
  • Method for predicting biological systems responses in hepatocytes
  • Method for predicting biological systems responses in hepatocytes

Examples

Experimental program
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preparation example Construction

[0045] Multiple markers can be combined in the preparation of a single sample to provide detection of multiple features in each individual hepatocyte in the population as well as in the entire population (Zhang et al., Cell, 2004.119(1): p.137 -44; Taylor et al., Drug Discov. Today, 2005.2(2): p.149-154). Quantum dots with a single stimulus wavelength and a narrow emission band offer the possibility of achieving a higher degree of diversity in testing (Michalet, et al., Science, 2005.307(5709): p.538-44) . In addition to fluorescent probes of various colors, a variety of bioluminescent and chemiluminescent reagents can also be effectively used in cell-based assays (Hemmila et al., J Fluoresc, 2005, 15(4): p.529- 42; Roda et al., Trends Biotechnol, 2004.22(6): p.295-303).

[0046] The hepatocytes can be plated on a substrate, such as a microplate, microscope slide, or other laboratory dish typically used for cell-based assays. Additionally, hepatocytes can be maintained on f...

Embodiment 1

[0071] This example demonstrates an embodiment of the invention in which a panel of test functional classes is used to create a profile of substances producing hepatotoxicity.

[0072] The functional categories to be tested for toxicity include stress pathways, mitochondrial function, cell cycle phase, morphological changes, apoptosis, nuclear changes, phospholipids, and peroxisome proliferation. In one embodiment, said functional class to be tested for toxicity is selected from the group consisting of mitochondrial function, apoptosis, nuclear alteration, phospholipidation, steatosis and DNA damage. In a preferred embodiment, at least one of said functional classes to be tested for toxicity is phospholipidation or steatosis. In another preferred embodiment, at least two functional classes are tested for toxicity, at least one of which is phospholipidation or steatosis. In another preferred embodiment, at least three functional classes are tested for toxicity, at least one of...

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Abstract

The inventive method employs a ''systems biology'' approach to predicting hepatotoxicity and other biological hepatocyte responses resulting from exposure to the test substance. In one embodiment, the invention provides an automated method for predicting the hepatocyte biological systems effect of a test substance. In another embodiment, the invention provides a method for constructing a knowledgebase (or database) of cellular systems biology response profiles for reference substances with known biological systems effects. In another embodiment, the invention provides a set of protocols and software tools used to carry out the profiling. Another embodiment of the invention is a panel of reagents. In another embodiment, the invention provides a set of protocols and software tools used to carry out the profiling. Another embodiment of the invention is a panel of reagents and protocols required for generating cellular systems biology response profiles, either to create a knowledgebase, or to use with an existing knowledgebase and informatics software to profile substance physiological effects. Another embodiment of the invention is a database of physiological profiles.

Description

[0001] related application [0002] This application claims priority to US Provisional Application No. 60 / 946,186, filed June 26, 2007. [0003] The aforementioned application is hereby incorporated by reference in its entirety. Background technique [0004] Basically, many drug candidates fail because of hepatocellular toxicity found in drug safety trials in animals or in late-stage clinical trials in humans. However, even after a drug is approved, there remains a substantial risk of hepatotoxicity in marketed drugs. Although there are occasional drug withdrawals not due to hepatotoxicity, liver toxicity is the most common reason for withdrawals, significantly limiting the use of approved drugs. In conclusion, hepatotoxicity can lead to futility and generate unnecessary costs that can be reduced by using early detection with high predictive value to identify hepatotoxicity in vivo. [0005] Considering the high sensitivity of the human liver to various environmental toxica...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50
CPCG01N33/5067
Inventor W·欧文L·维尔内帝P·A·约翰斯顿D·L·泰勒
Owner CELLUMEN INC
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