TOXICITY TYPING USING LIVER STEM CELLS

• US 20120220469A1
• Filed: 02/21/2012
• Published: 08/30/2012
• Est. Priority Date: 06/14/2000
• Status: Active Grant

• Current Assignee: VistaGen Therapeutics, Inc.
• Original Assignee: VistaGen Therapeutics, Inc.
• Inventors: SNODGRASS, H. Ralph
• Granted Patent: US 8,512,957 B2
• Time in Patent Office: Days
• US Class Current: 506/6
• CPC Class Codes: C12Q 1/6809 Methods for determination or identification of nucleic acids involving differential detection
• CPC Class Codes: G01N 33/5014 for testing toxicity

Abstract

This invention provides methods and systems for identifying and typing toxicity of chemical compositions, as well as for screening new compositions for toxicity. The invention involves detecting alterations in gene or protein expression and hence establishing molecular profiles in isolated mammalian LSCs contacted with various chemical compositions of known and unknown toxicities, and correlating the molecular profiles with toxicities of the chemical compositions.

Claims

1. A method of creating a molecular profile of a chemical composition, comprising the steps of:

a) contacting an isolated population of mammalian liver stem cells (LSCs) with the chemical composition; and

b) recording alterations in protein expression in the mammalian LSCs in response to the chemical composition to create a molecular profile of the chemical composition.

3. The method of claim 1 or 2, wherein levels of protein expression are detected by a label.

4. The method of claim 3, wherein the label is selected from the group consisting of a fluorescent, a colorimetric, a radioactive, an enzyme, an enzyme substrate, a nucleoside analog, a magnetic, a glass, a latex bead, a colloidal gold, and an electronic transponder.

7. :

The method of claim 1 or 2, wherein the molecular profile comprises alterations in protein expression.

8. The method of claim 7, wherein the alterations in protein expression are detected by an immunoactivity assay.

9. The method of claim 7, wherein the alterations in protein expression are detected by a mass spectrometry assay.

19. A library of molecular profiles of chemical compositions having predetermined toxicities, produced by a method according to any one of the claims 2 or 10-18, wherein the library is in a computer readable media and a computer containing the library is used in a method of typing toxicity of a test chemical composition, comprising the steps of:

a) creating a molecular profile of the test chemical composition according to claim 1; and

b) comparing the molecular profile in step a) with the library profile of chemical compositions having predetermined toxicities;

wherein the type of toxicity of the test chemical composition is determined by the comparison in step b).

20. The library of claim 19, wherein the library comprises molecular profiles for at least 20 chemical compositions.

21. A method of typing toxicity of a test chemical composition, comprising the steps of:

a) creating a molecular profile of the test chemical composition according to claim 1; and

b) comparing the molecular profile in step a) with the molecular profile of a chemical composition having predetermined toxicities;

wherein the type of toxicity of the test chemical composition is determined by the comparison in step b).

22. A systematic method of typing toxicity of a test chemical composition, comprising the steps of:

a) creating a molecular profile of the test chemical composition according to claim 1; and

b) comparing the molecular profile in step a) with a composite library of molecular profiles of chemical compositions having predetermined toxicities, wherein the composite library comprises the molecular profiles of at least two chemical compositions, said molecular profiles are created according to claim 1;

wherein the type of toxicity of the test chemical composition is determined by the comparison in step b).

23. A method of ranking toxicity of a test chemical composition, the method comprising:

a) creating a molecular profile of the test chemical composition according to claim 1; and

b) comparing the molecular profile in step a) with a composite library of molecular profiles of chemical compositions having predetermined toxicities, wherein the composite library comprises the molecular profiles of at least two chemical compositions, said molecular profiles are created according to claim 1;

wherein the toxicity of the test chemical composition is ranked by the comparison in step b).

24. The method of claim 21, 22 or 23, wherein the test chemical composition is known or unknown.

25. The method of claim 21, 22 or 23, further wherein the LSCs are of human origin.

26. The method of claim 25, further wherein the chemical compositions having predetermined toxicities are therapeutic agents, neurotoxins, renal toxins, hepatic toxins, toxins of hematopoietic cells, or myotoxins.

27. The method of claim 25, further wherein the chemical compositions having predetermined toxicities are selected from the group consisting of agents that are toxic to cells of one or more reproductive organs, teratogenic agents and carcinogens.

28. The method of claim 25, further wherein the chemical compositions having predetermined toxicities are selected from the group consisting of agricultural chemicals, cosmetics, and environmental contaminants.

29. The method of claim 21, 22 or 23, further wherein the LSCs are of non-human mammals.

30. The method of claim 29, wherein the non-human mammals are rodents.

31. The method of claim 29, further wherein the chemical compositions having predetermined toxicities are selected from the group consisting of animal therapeutics, neurotoxins, renal toxins, hepatic toxins, toxins of hematopoietic cells, and myotoxins.

32. The method of claim 29, further wherein the chemical compositions having predetermined toxicities are selected from the group consisting of agents that are toxic to cells of one or more reproductive organs, teratogenic agents and carcinogens.

33. The method of claim 29, further wherein the chemical compositions having predetermined toxicities are selected from the group consisting of agricultural chemicals, cosmetics, and environmental contaminants.

2. A method of compiling a library of molecular profiles of chemical compositions having predetermined toxicities, comprising the steps of:

a) contacting an isolated population of mammalian liver stem cells (LSCs) with a chemical composition having predetermined toxicities;

b) recording alterations in protein expression in the mammalian LSCs in response to the chemical composition to create a molecular profile of the chemical composition; and

c) compiling a library of molecular profiles by repeating steps a) and b) with at least two chemical compositions having predetermined toxicities.

10. The method of claim 2, wherein the LSCs are of human origin.

11. The method of claim 10, further wherein the chemical compositions having predetermined toxicities are selected from the group consisting of therapeutic agents, neurotoxins, renal toxins, hepatic toxins, toxins of hematopoietic cells, and myotoxins.

12. The method of claim 10, further wherein the chemical compositions having predetermined toxicities are selected from the group consisting of agents that are toxic to cells of one or more reproductive organs, teratogenic agents and carcinogens.

13. The method of claim 10, further wherein the chemical compositions having predetermined toxicities are selected from the group consisting of agricultural chemicals, cosmetics, and environmental contaminants.

14. The method of claim 2, wherein the LSCs are of non-human mammals.

15. The method of claim 14, wherein the non-human mammals are rodents.

16. The method of claim 14, further wherein the chemical compositions having predetermined toxicities are selected from the group consisting of animal therapeutics, neurotoxins, renal toxins, hepatic toxins, toxins of hematopoietic cells, and myotoxins.

17. The method of claim 14, further wherein the chemical compositions having predetermined toxicities are selected from the group consisting of agents that are toxic to cells of one or more reproductive organs, teratogenic agents and carcinogens.

18. The method of claim 14, further wherein the chemical compositions having predetermined toxicities are selected from the group consisting of agricultural chemicals, cosmetics, and environmental contaminants.

5-6. -6. (canceled)

34-41. -41. (canceled)