Method of diagnosing of exposure to toxic agents by measuring distinct pattern in the levels of expression of specific genes

• US 20030044790A1
• Filed: 06/07/2001
• Published: 03/06/2003
• Est. Priority Date: 02/05/1999
• Status: Abandoned Application

• Current Assignee: Chanaka Mendis, Marti Jett, Rina Das
• Original Assignee: Chanaka Mendis, Marti Jett, Rina Das
• Inventors: Mendis, Chanaka, Das, Rina, Jett, Marti
• Application Number:
  US09/876,249
• Publication Number:
  US 20030044790A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C12N 15/1089 Design, preparation, screening or analysis of libraries using computer algorithms
• CPC Class Codes: G01N 33/6803 General methods of protein analysis not limited to specific proteins or families of proteins

Abstract

A method of diagnosing exposure to a toxic agent comprising the steps of detecting the amount of protein/gene expression present in a sample of mammalian tissue or mammalian body fluids that has not been exposed to a toxic agent. Then the amount of protein/gene expression present in a sample of mammalian tissue or mammalian body fluids that has been exposed to the toxic agent is detected. A determination of the difference in the detected amount of protein/gene expression between the exposed and unexposed samples is made. A comparison of the difference to a library of expected protein/gene expression for predetermined toxic agents is made. Finally, an evaluation is made whether the difference indicates the exposure to a particular toxic agent. A treatment method for administering a therapeutic agent which inhibits the mechanistic pathways necessary to maintain the progression of lethal shock is also disclosed.

Claims

1. A method of pre-symptom diagnosis of exposure to a toxic agent comprising:

obtaining a sample of mammalian body fluid or tissue from a subject;

detecting a pattern of gene expression or protein expression present in said sample;

comparing the pattern of gene or protein expression from said sample with a library of known patterns of gene or protein expressions for toxic agents; and

determining whether the subject has been exposed to a toxic agent and if said subject has been exposed to a toxic agent, identifying said toxic agent based on information in said library.

2. The method of claim 1, wherein said sample comprises peripheral blood lymphoid cells.

3. The method of claim 1, wherein said sample comprises mammalian tissue.

5. The method of claim 1, wherein said toxic agent is SEB.

6. The method of claim 1, wherein said toxic agent is anthrax.

7. The method of claim 1, wherein said toxic agent is chorera.

8. The method of claim 1, wherein said toxic agent is Brucella.

9. The method of claim 1, wherein said toxic agent is plague.

10. The method of claim 1, wherein said toxic agent is botulinum toxin.

11. The method of claim 1, wherein said library comprises gene and protein patterns for SEB, LPS, anthrax, cholera, Brucella, plague and botulinum toxin.

20. A method of treating impending illness from exposure to toxic agents based on identified decrements or excesses of expressed genes which are responsible and/or lead to illness caused by the toxic agent, comprising the steps of:

a) diagnosing a presence of and type of toxic agent according to the method of claim 1;

and

b) administering a therapeutic agent that blocks the effects of said toxic agents.

21. The method of claim 20, wherein said therapeutic agent is HPA-Na.

22. The method of claim 20, wherein said therapeutic agent is P-38.

23. The method of claim 20, wherein said therapeutic agent is a protein kinase C inhibitor selected from the group consisting of H-7 and Chelerythrine.

24. A method of pre- symptom treatment of lethal shock induced by toxic agents comprising the steps of:

a) Detecting the presence of and type of toxic agent according to the method of claim 1;

and

b) administering a therapeutic agent which inhibits gene/protein expression necessary to maintain the progression of lethal shock.

4. The method of claim ., wherein said toxic agent is LPS.

12. A method of diagnosing exposure to a toxic agent comprising the steps of:

detecting an amount of protein or gene expression present in a sample of mammalian tissue or mammalian body fluids that has been unexposed to the toxic agent;

detecting an amount of protein or gene expression present in a sample of mammalian tissue or mammalian body fluids has been exposed to the toxic agent;

determining the difference in the detected amount of protein/gene expression between the exposed and unexposed samples;

comparing the difference to a library of expected protein/gene expression for predetermined toxic agents;

evaluating whether the difference indicates that said sample has been exposed to a particular toxic agent.

13. The method of claim 12, further comprising a step of identifying said toxic agent.

14. The method of claim 12, wherein said library of toxic agents comprises expected protein or gene expression for toxic agents selected from the group consisting of SEB, LPS, anthrax, Brucella, plague, botulinum toxin, and cholera toxin.

15. The method of claim 12, wherein said expression is a gene expression.

16. The method of claim 12, wherein said expression is a protein expression.

17. A method of diagnosing exposure to a toxic agent comprising the steps of:

detecting an amount of protein/gene expression present in a sample of mammalian tissue or mammalian body fluids that has been unexposed to the toxic agent; and

screening for specific mRNA induced by toxic agents for diagnostic markers for early indication of impending illness.

18. A method of diagnosing exposure to a toxic agent comprising the steps of:

detecting patterns of gene expression or proteins present in a sample of mammalian tissue or mammalian body fluids from persons that have been exposed to potentially toxic agent(s);

determining a relative amount of expression of a panel of genes relative to house keeping genes expressed in said tissue or fluids from the potentially exposed persons;

comparing the differences in the relative amounts to a library of expected gene expression or proteins for predetermined toxic agents; and

evaluating whether the differences indicate that exposure has occurred to a known, catalogued, toxic agent, to a previously unknown toxic agent, or to a toxic agent mixed with potentiating agents.

19. A method of evaluation of impending illness prior to appearance of symptoms, for triage or for determination of time, extent or degree of exposure by screening for specific mRNA induced by toxic agents for diagnostic markers comprising the steps of:

a) detecting patterns of gene expression or proteins present in a sample of mammalian tissue or mammalian body fluids from persons that have been exposed to potentially toxic agent(s);

b) determining a relative amount of expression of a panel of genes relative to housekeeping genes expressed in said tissues or fluids from the potentially exposed persons;

c) comparing the relative amount differences to a library of expected gene expression/proteins for predetermined toxic agents;

d) evaluating whether the differences indicates that exposure has occurred to a known, catalogued, toxic agent, to a previously unknown toxic agent, or to a toxic agent mixed with potentiating agents.

25. A method of treatment with drugs/inhibitors of lethal shock induced by toxic agents comprising the steps of:

a) determining mechanisms contributing to lethal shock based on gene expression in mammalian samples exposed to the toxic agent(s);

b) administering a therapeutic agent which inhibits pathways of the mechanisms necessary to maintain the progression of lethal shock.

26. A method of treating impending illness in a mammalian subject from exposure to toxic agents based on identified decrements or excesses of expressed genes which are responsible and/or lead to illness caused by the toxic agent, comprising the steps of:

detecting patterns of gene expression or proteins present in a sample of mammalian tissue or mammalian body fluids from said mammalian subject that has been exposed to potentially toxic agent(s);

determining a relative amount of expression of a panel of genes relative to housekeeping genes expressed in those tissues from the mammalian subject;

comparing the relative amount differences to a library of expected gene expression/proteins for predetermined toxic agents;

evaluating whether the differences indicate that exposure has occurred to a known, catalogued, toxic agent, to a previously unknown toxic agent, or to a toxic agent mixed with potentiating agents; and

treating the mammalian subject with a therapeutic agent which blocks the gene expression leading to the progression of toxic induces illness.

27. The method of claim 26, wherein said therapeutic agent is P-38.

28. The method of claim 26, wherein said therapeutic agent is HPA-Na.

29. The method of claim 26, wherein said therapeutic agent targets specific genes.

30. The method of claim 26, wherein said therapeutic agent is an antisense.