Method for detection of cancer based on spatial genome organization
View Patent ↗The invention provides methods of detecting abnormal cells in a sample using the radial position of one or more genes within the nucleus of a cell, as well as a kit for detecting abnormal cells using such methods. The invention also provides methods of identifying gene markers for abnormal cells using the radial position of one or more genes within the nucleus of a cell.
1. A method of detecting a peripheral position of the HES5 gene in cells in a breast tissue sample from a human subject, the method comprising:
(a) obtaining a breast tissue sample comprising one or more cells from the subject, wherein one or more cells in the breast tissue sample has a nucleus, and wherein each nucleus has a nuclear center and a nuclear membrane;
(b) contacting the breast tissue sample with a labeled DNA probe;
(c) hybridizing the labeled DNA probe with the HES5 gene by fluorescence in situ hybridization (FISH);
(d) providing a positive and/or negative control; and
(e) detecting a peripheral position of the HES5 gene within the nucleus relative to the nuclear center and/or nuclear membrane by FISH with the labeled DNA probe, wherein the peripheral position of the HES5 gene is:
(i) a statistical difference between the position of the HES5 gene in the breast tissue sample and the nuclear center position of the HES5 gene in the negative control; and/or
(ii) a lack of a statistical difference between the position of the HES5 gene in the breast tissue sample and the peripheral position of the HES5 gene in the positive control.
2. The method of claim 1 , further comprising detecting a peripheral position of one or more of HSP90AA1, FOSL2, CSF1R, MYC, TGFB3, ERBB2, and AKT1 genes within the nucleus relative to the nuclear center and/or nuclear membrane by FISH, wherein the peripheral position is:
(i) a statistical difference between the position of one or more of the HSP90AA1, FOSL2, CSF1R, MYC, TGFB3, ERBB2, and AKT1 genes in the breast tissue sample and the nuclear center position of the respective one or more genes in the negative control; and/or
(ii) a lack of a statistical difference between the position of one or more of the HSP90AA1, FOSL2, CSF1R, MYC, TGFB3, ERBB2, and AKT1 genes in the breast tissue sample and the peripheral position of the respective one or more genes in the positive control.
3. The method of claim 2 , further comprising
detecting an increased proximity to the nuclear membrane of (i) the HES5 gene and (ii) one or more of the HSP90AA1, FOSL2, CSF1R, MYC, TGFB3, ERBB2, and AKT1 genes in the breast tissue sample relative to the proximity to the nuclear membrane of (i) the HES5 gene and (ii) the respective one or more genes in the negative control.
4. The method of claim 2 , wherein the breast tissue sample is obtained using a needle biopsy.
5. The method of claim 2 , wherein the positive control is a standardized distribution profile of the position of (i) the HES5 gene and (ii) one or more of the HSP90AA1, FOSL2, CSF1R, MYC, TGFB3, ERBB2, and AKT1 genes in a breast cancer control sample and the negative control is a standardized distribution profile of the position of (i) the HES5 gene and (ii) one or more of the HSP90AA1, FOSL2, CSF1R, MYC, TGFB3, ERBB2, and AKT1 genes in a non-cancerous control sample from breast tissue.
6. The method of claim 2 , comprising detecting the positions within the nucleus relative to the nuclear center and/or nuclear membrane of the following:
(a) HES5 and HSP90AA1,
(b) HES5 and FOSL2,
(c) HES5 and CSF1R, or
(d) HES5 and MYC.
7. The method of claim 2 , comprising detecting the positions within the nucleus relative to the nuclear center and/or nuclear membrane of the following:
(a) HES5 and TGFB3,
(b) HES5 and ERBB2, or
(c) HES5 and AKT1.
8. The method of claim 2 , comprising detecting the positions within the nucleus relative to the nuclear center and/or nuclear membrane of (i) the HES5 gene and (ii) two or more of the HSP90AA1, FOSL2, CSF1R, MYC, TGFB3, ERBB2, and AKT1 genes by FISH.
9. The method of claim 8 , comprising detecting the positions within the nucleus relative to the nuclear center and/or nuclear membrane of the HES5, TGFB3, and FOSL2 genes.
10. The method of claim 8 , wherein the statistical difference between the positions of (i) the HES5 gene and one or more of the HSP90AA1, FOSL2, CSF1R, MYC, TGFB3, ERBB2, and AKT1 genes in the breast tissue and (ii) the nuclear center positions of the HES5 gene and the respective one or more genes in the negative control is a P-value of P<0.01.
11. The method of claim 2 , wherein the statistical difference between the positions of (i) the HES5 gene and one or more of the HSP90AA1, FOSL2, CSF1R, MYC, TGFB3, ERBB2, and AKT1 genes in the breast tissue sample and (ii) the nuclear center positions of the HES5 genes and the respective one or more genes in the negative control is a P-value of P<0.01.
12. The method of claim 2 , comprising detecting an increased proximity to the nuclear center of (i) the HES5 gene and (ii) one or more of the HSP90AA1, FOSL2, CSF1R, MYC, TGFB3, ERBB2, and AKT1 genes in the breast tissue sample relative to the proximity to the nuclear membrane of (i) the HES5 gene and the (ii) the respective one or more genes in the positive control.
13. The method of claim 1 , further comprising
detecting an increased proximity to the nuclear membrane of the HES5 gene in the breast tissue sample relative to the proximity to the nuclear membrane of the HES5 gene in the negative control.
14. The method of claim 1 , wherein the breast tissue sample is obtained using a needle biopsy.
15. The method of claim 1 , wherein the positive control is a standardized distribution profile of the position of the HES5 gene in a breast cancer control sample and the negative control is a standardized distribution profile of the position of the HES5 gene in a non-cancerous control sample from breast tissue.
16. The method of claim 1 , wherein the statistical difference between the position of the HES5 gene in the breast tissue sample and the nuclear center position of the HES5 gene in the negative control is a P-value of P<0.01.
17. The method of claim 1 , further comprising treating cancer in the subject.
18. The method of claim 1 , comprising detecting an increased proximity to the nuclear center of the HES5 gene in the breast tissue sample relative to the proximity to the nuclear membrane of the HES5 gene in the positive control.