Determination of molecular age by detection of
View Patent ↗The presently disclosed subject matter provides methods for determining the molecular age of a tissue. Also provided are methods for employing a molecular age that has been determined for a tissue for assessing the appropriateness of various therapeutic applications for the tissue or a subject comprising the tissue.
1. A method for predicting a tendency to therapy-induced toxicity in a subject, the method comprising:
(a) providing a subject under consideration for a therapy;
(b) providing a sample from the subject, wherein the sample comprises a cell or tissue having a molecular age;
(c) determining the molecular age of the cell or tissue by:
(i) quantifying a level of expression of p16 INK4a , ARF, or both p16 INK4a and ARF in the sample;
(ii) comparing the level of expression quantified with a standard, wherein the comparing comprises determining whether an increased level of expression of p16 INK4a , ARF, or both p16 INK4a and ARF is present or absent in the sample relative to the standard; and
(iii) determining an increased molecular age if an increased level is present or a lack of an increased molecular age if an increased level is absent; and
(d) predicting an increased tendency to therapy-induced toxicity in the subject if an increased molecular age is present or a decreased tendency to therapy-induced toxicity in the subject if an increased molecular age is absent.
2. The method of claim 1 , further comprising treating the subject with the therapy if a decreased tendency to therapy-induced toxicity is predicted or not treating the subject with the therapy if an increased tendency to therapy-induced toxicity is predicted.
3. The method of claim 1 , further comprising treating the subject with the therapy and one or more additional supportive treatments if an increased tendency to therapy-induced toxicity is predicted.
4. A method for determining suitability of a cell or tissue for use in a donation, the method comprising:
(a) providing a sample from the cell or tissue, wherein the cell or tissue has a molecular age;
(b) determining the molecular age of the cell or tissue by:
(i) quantifying a level of expression of p16 INK4a , ARF, or both p16 INK4a and ARF in the sample;
(ii) comparing the level of expression quantified with a standard, wherein the comparing comprises determining whether an increased level of expression of p16 INK4a , ARF, or both p16 INK4a and ARF is present or absent in the sample relative to the standard; and
(iii) determining the molecular age based on whether an increased level is present or absent; and
(c) determining a suitability of the cell or tissue for use in a donation based on the molecular age.
5. The method of claim 2 , wherein the cell or tissue donation is appropriate for a transplant selected from the group consisting of bone marrow transplant, solid organ transplant, and tissue graft transplant.
6. A method for determining the molecular age of a cell or tissue in a subject, the method comprising:
a) providing a sample from the cell or tissue;
b) quantifying a level of expression of p16 INK4a , ARF, or both p16 INK4a and ARF in the sample; and
c) determining the molecular age of the cell or tissue by comparing the level of expression of p16 INK4a , ARF, or both p16 INK4a and ARF to a standard.
7. The method of claims 1 , 4 , or 6 , wherein the sample comprises a cell or tissue selected from the group consisting of an adrenal gland cell, a bone marrow cell, a cecum cell, a brain cortex cell, a duodenum cell, a heart cell, a kidney cell, a liver cell, a lung cell, a lymph node cell, an ovary cell, a pancreas cell, a spleen cell, a testis cell, a blood cell, and a uterus cell.
8. The method of claims 1 , 4 , or 6 , further comprising purifying a particular cell type from tissue.
9. The method of claim 5 , wherein the particular cell type comprises a lymphocyte, a stem cell, a neural cell, and an islet cell.
10. The method of claims 1 , 4 or 6 , wherein the standard comprises a quantification of a level of expression of p16 INK4a , ARF, or both p16 INK4a and ARF in a cell or tissue in a population of same species individuals as the subject.
11. The method of claims 1 , 4 , or 6 , wherein the standard comprises a quantification of a level of expression of p16 INK4a , ARF, or both p16 INK4a and ARF in a cell or tissue in the subject at an earlier time.
12. The method of claims 1 , 4 , or 6 , wherein the standard comprises a quantification of a level of expression of p16 INK4a , ARF, or both p16 INK4a and ARF in a second sample isolated from a different tissue of the subject.
13. The method of claim 9 , wherein the at least one different tissue comprises a cell selected from the group consisting of an adrenal gland cell, a bone marrow cell, a cecum cell, a brain cortex cell, a duodenum cell, a heart cell, a kidney cell, a liver cell, a lung cell, a lymph node cell, an ovary cell, a pancreas cell, a spleen cell, a testis cell, a blood cell, and a uterus cell.
14. The method of claims 1 , 4 , or 6 , wherein the subject is a human.
15. The method of claims 1 , 4 , or 6 , wherein the quantifying employs quantitative real-time polymerase chain reaction analysis.
16. The method of claims 1 , 4 , or 6 , wherein the comparing the level comprises determining an increased level of expression of p16 INK4a , ARF, or both p16 INK4a and ARF, and the increased level of expression indicates increased molecular age of the tissue relative to the standard.
17. The method of claims 1 , 4 , or 6 , wherein the comparing is to a level of expression of p16 INK4a , ARF, or both p16 INK4a and ARF in the target cell type in a population of same species individuals as the subject.
18. The method of claims 1 , 4 , or 6 , further comprising normalizing the level of expression quantified to a level of expression of a housekeeping gene.