Diagnosis of diseases and conditions by analysis of histopathologically processed biological samples using liquid tissue preparations
The invention provides methods for diagnosing diseases such as cancer and other conditions using biological samples. Liquid Tissue samples prepared from histopathologically prepared tissue obtained from a subject surprisingly can be used to identify and, optionally, to quantify analytes that are diagnostic of the presence of a disease, condition or syndrome in the subject.
1. A method of diagnosing the presence of a disease, condition, or syndrome in a subject, comprising: detecting and quantitating the presence of at least one protein analyte in a preparation from a biological sample obtained from a subject suspected of suffering from a disease, condition, or syndrome, wherein the biological sample comprises formalin-fixed tissue or paraffin-embedded tissue, wherein the presence of said analyte indicates that the subject has the disease, condition, or syndrome; and wherein said preparation from a biological sample is prepared by a method comprising the steps of:
(a) heating a composition comprising a histopathologically processed biological sample and a reaction buffer at a temperature from 80° C. to 100° C. for a period of time from 10 minutes to 4 hours to negatively affect protein crosslinking in said biological sample, and
(b) treating the resulting composition with an effective amount of a proteolytic enzyme for a time sufficient to disrupt the tissue and cellular structure of said biological sample and to liquefy said sample, thereby producing a liquid, soluble, dilutable biomolecule lysate that is representative of the total protein content of said histopathologically processed biological sample
wherein said quantitating comprises the addition of AQUA peptides to said biological sample.
2. The method of claim 1 , wherein the detection comprises mass spectrometry, immunohistochemistry, ELISA, Western blotting, polyacrylamide gel electrophoresis, synthetic aptamer binding analysis, or isoelectric focusing analysis.
3. The method of claim 1 , wherein said preparation is in the form of an array.
4. The method of claim 1 , wherein said subject is human.
5. The method of claim 1 , wherein said protein analyte is not substantially cross-linked.
6. The method of claim 1 , further comprising comparing the one or more detected analytes with one or more reference analytes obtained from a subject known to have or not have the disease, condition, or syndrome.
7. The method of claim 2 , wherein the detecting is by a mass spectrometric selected from the group consisting of matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF), liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS), or surface enhanced laser desorption ionization (SELDI) mass spectrometry.
8. The method of claim 1 , wherein said analyte is selected from the group consisting of the analytes listed in Table 1.
9. A method of detecting cancer in a subject, comprising: detecting and quantitating the presence of at least one protein analyte in a preparation from a biological sample obtained from a subject suspected of suffering from a disease, condition, or syndrome, wherein the biological sample comprises formalin-fixed tissue or paraffin-embedded tissue, wherein the presence of said analyte indicates that the subject has cancer; and wherein said preparation from a biological sample is prepared by a method comprising the steps of:
(a) heating a composition comprising a histopathologically processed biological sample and a reaction buffer at a temperature from 80° C. to 100° C. for a period of time from 10 minutes to 4 hours to negatively affect protein crosslinking in said biological sample, and
(b) treating the resulting composition with an effective amount of a proteolytic enzyme for a time sufficient to disrupt the tissue and cellular structure of said biological sample producing a liquid, soluble, dilutable biomolecule lysate that is representative of the total protein content of said histopathologically processed biological sample
wherein said quantitating comprises the addition of AQUA peptides to said biological sample.
10. The method of claim 9 , wherein said analyte is a peptide derived from Her2, an estrogen receptor, a progesterone receptor, PSA or an androgen receptor.
11. The method of claim 9 , wherein the cancer is prostate cancer and the analyte is a peptide derived from PSA, PAP, PEBP or GDFI5.
12. The method of claim 9 , further comprising comparing the protein analytes with one or more reference analytes obtained from a subject known to have or not have the disease, condition, or syndrome.
13. A method of determining the predisposition to develop a disease or condition in a subject, comprising:
(a) detecting the presence of at least one protein analyte in a preparation from a biological sample obtained from a subject suspected of suffering from a disease, condition, or syndrome, wherein the biological sample comprises formalin-fixed tissue or paraffin-fixed tissue, wherein said preparation from a biological sample is prepared by a method comprising the steps of:
(i) heating a composition comprising a histopathologically processed biological sample and a reaction buffer at a temperature from 80° C. to 100° C. for a period of time from 10 minutes to 4 hours to negatively affect protein crosslinking in said biological sample, and
(ii) treating the resulting composition with an effective amount of a proteolytic enzyme for a time sufficient to disrupt the tissue and cellular structure of said biological sample producing a liquid, soluble, dilutable biomolecule lysate that is representative of the total protein content of said histopathologically processed biological sample; and
(b) comparing the one or more detected protein analytes with one or more reference analytes obtained from a subject known to have the disease or condition, wherein the presence of the one or more analytes indicates that the subject has an increased predisposition to develop the disease or condition as compared to a subject not having said one or more analytes;
wherein said quantitating comprises the addition of AQUA peptides to said biological sample.
14. The method of claim 13 , wherein the disease is cancer and the analyte is a peptide derived from Her2, an estrogen receptor, a progesterone receptor, PSA or an androgen receptor.
15. The method of claim 13 , wherein the disease is Alzheimer disease and the analyte is a peptide derived from CSF beta-amyloid protein or tau protein.
16. The method of claim 15 , wherein the disease is Parkinson's disease and the analyte is a peptide derived from alpha synuclein, parkin, ubiquitin C-terminal hydrolase L (UCH-L1), or NR4A2.
17. The method of claim 13 , wherein the condition is a benign hyperplastic condition.