Methods of treating a subject with a high gleason score prostate cancer
The present invention relates generally to the field of biomarker analysis, particularly determining gene expression signatures from urine samples. The disclosure provides compositions, kits and methods for diagnosing a prostate disorder such as prostate cancer in a male subject.
1. A method of treating a subject with a high risk for a high Gleason score prostate cancer, wherein the high Gleason score prostate cancer has a Gleason score of greater than 6, the method comprising the steps of:
a. extracting one or more mRNAs from a urine sample from the subject;
b. detecting the level of mRNA expression of PCA3, ERG and SPDEF;
c. normalizing the level of mRNA expression of PCA3 and ERG to SPDEF;
d. computing an EXO106 score using the formula:
(
log
2
max
(
1
,
ERG
copies
)
SPDEF
copies
+
log
2
max
(
1
,
PCA
3
copies
)
SPDEF
copies
+
16.92
)
*
1.83
,
wherein ERG copies is the level of mRNA expression of ERG, PCA3 copies is the level of mRNA expression of PCA3 and SPDEF copies is the level of SPDEF mRNA expression;
e. comparing the EXO106 Score to a predetermined cutoff value; and
f. treating the subject at a high risk for a high Gleason score prostate cancer when the EXO106 score is greater than the predetermined cutoff value.
2. The method of claim 1 , wherein the urine sample is the first 40 mL voided from the bladder of the subject.
3. The method of claim 1 , wherein the urine sample is the first 20 mL voided from the bladder of the subject.
4. The method of claim 1 , wherein the predetermined cutoff value is 10.
5. The method of claim 1 , wherein extracting one or more mRNAs in step (a) comprises:
a. isolating a microvesicle fraction from the urine sample; and
b. extracting one or more mRNAs from the microvesicle fraction.
6. The method of claim 5 , wherein the step of isolating the microvesicle fraction comprises processing the sample to remove cells and cell debris and concentrating the microvesicle fraction by exposing the microvesicle fraction to ultrafiltration or a filtration concentrator, and washing the microvesicle fraction prior to extracting the one or more nucleic acids from the microvesicle fraction.
7. The method of claim 6 , wherein the method further comprises adding an RNase inhibitor to the microvesicle fraction prior to extracting the one or more nucleic acids from the microvesicle fraction.