METHODS TO ASSESS THE LIKELIHOOD OF DYSPLASIA OR ESOPHAGEAL ADENOCARCINOMA
In some embodiments, a method for aiding assessment of the likelihood of dysplasia or esophageal adenocarcinoma being present in a subject can include (a) providing an esophagal sample from said subject (b) determining the methylation status of (i) SLC22A18, (ii) PIGR, (iii) GJA12 and (iv) RIN2 in said sample wherein if 2 or more of said genes are methylated then an increased likelihood of presence of dysplasia or esophageal is determined. The invention also relates to apparatus for same.
1 . A method for aiding assessment of the likelihood of dysplasia or esophagal adenocarcinoma being present in a subject, the method comprising
(a) providing an esophagal sample from said subject
(b) determining the methylation status of
(i) SLC22A18,
(ii) PIGR,
(iii) GJA12 and
(iv) RIN2
in said sample
wherein if 2 or more of said genes are methylated then an increased likelihood of presence of dysplasia or esophagal adenocarcinoma is determined.
2 . A method according to claim 1 wherein the method further comprises determining the methylation status of (v) TCEAL7.
3 . A method according to claim 1 wherein if said subject is male, the method further comprises determining the methylation status of (vi) RGN.
4 . A method according to claim 1 wherein the dysplasia is high grade dysplasia (HGD).
5 . A method of assessing the risk for a particular subject comprising performing the method according to claim 1 , wherein if 0 or 1 of said genes are methylated then low risk is determined, and if 2 of said genes are methylated then intermediate risk is determined, if 3 or more of said genes are methylated then high risk is determined.
6 . A method according to claim 1 wherein methylation status is determined by pyrosequencing.
7 . A method according to claim 6 wherein said pyrosequencing is carried out using one or more sequencing primers selected from Supplementary Table 5.
8 . A method according to claim 1 wherein the methylation status is scored by determining the percentage methylation of each of said genes and comparing the values to the following methylation cut off percentages:
Gene Methylation cut-off (%)
GJA12 51.74000
SLC22A18 49.25000
PIGR 64.755000
RIN2 37.85500
RGN (males only) 18.645000
TCEAL7 58.54000
wherein a value for a gene which exceeds the methylation cut off percentage for said gene is scored as ‘methylated’.
9 . An apparatus or system which is
(a) configured to analyse an esophagal sample from a subject, wherein said analysis comprises
(b) determining the methylation status of
(i) SLC22A18,
(ii) PIGR,
(iii) GJA12 and
(iv) RIN2
in said sample,
said apparatus or system comprising an output module,
wherein if 2 or more of said genes are methylated then an increased likelihood of presence of dysplasia or esophagal adenocarcinoma is determined.
10 . An apparatus according to claim 9 wherein the analysis further comprises determining the methylation status of (v) TCEAL7.
11 . An apparatus according to claim 9 wherein if said subject is male, the analysis further comprises determining the methylation status of (vi) RGN.
12 . A method according to claim 1 wherein said sample comprises frozen biopsy material.
13 . A method for aiding assessment of the likelihood of dysplasia or esophagal adenocarcinoma being present in a subject, the method comprising
(a) providing an esophagal sample from said subject
(b) determining the methylation status of
(i) SLC22A18,
(ii) PIGR,
(iii) GJA12 and
(iv) RIN2
in said sample;
comparing the methylation values of (b) to a reference standard,
wherein if 2 or more of said genes are methylated at a level higher than the reference standard then an increased likelihood of presence of dysplasia or esophagal adenocarcinoma is determined.
14 . A method according to claim 13 wherein said reference standard is from a subject having Barrett's esophagus, but not having dysplasia or esophagal adenocarcinoma.
15 . A method according to claim 13 wherein said reference standard comprises columnar epithelium such as Barrett's esophagus or duodenum.
16 . A method according to claim 13 wherein the method further comprises determining the methylation status of (v) TCEAL7.
17 . A method according to claim 13 wherein if said subject is male, the method further comprises determining the methylation status of (vi) RGN.
18 . A computer program product operable, when executed on a computer, to perform the method steps of claim 1 .
19 . (canceled)