Gene expression profiles associated with sub-clinical kidney transplant rejection
By a genome-wide gene analysis of expression profiles of over 50,000 known or putative gene sequences in peripheral blood, the present inventors have identified a consensus set of gene expression-based molecular biomarkers associated with subclinical acute rejection (subAR). These genes sets are useful for diagnosis, prognosis, monitoring of subAR.
1. A method of treating a transplant recipient on an immunosuppressive drug comprising:
(a) obtaining nucleic acids of interest, wherein the nucleic acids of interest comprise mRNA derived from a blood sample from the transplant recipient or cDNA complements of mRNA derived from a blood sample from the transplant recipient wherein the transplant recipient has a serum creatinine level that is stable and less than 2.3 mg/dL;
(b) performing a microarray assay or Next Generation sequencing assay on the nucleic acids of interest obtained in (a) to detect expression levels of at least five genes, wherein the at least five genes are capable of specifically detecting subclinical acute rejection (subAR) in a transplant patient with stable serum creatinine levels;
(c) detecting subclinical acute rejection in the transplant recipient that has the serum creatinine level that is stable and less than 2.3 mg/dL based on the expression levels detected in (b); and
(d) administering a new immunosuppressive drug or a higher dose of the immunosuppressive drug to the transplant recipient in order to treat the subclinical acute rejection detected in (c).
2. The method of claim 1 , wherein the transplant recipient that has the serum creatinine level that is stable and less than 2.3 mg/dL also has normal serum creatinine levels.
3. The method of claim 1 , further comprising contacting the nucleic acids of interest with probes, wherein the probes are specific for the at least five genes selected in (b).
4. The method of claim 1 , wherein the method is repeated at different times on the transplant recipient that has the serum creatinine level that is stable and less than 2.3 mg/dL.
5. The method of claim 1 , wherein the transplant recipient that has the serum creatinine level that is stable and less than 2.3 mg/dL is a kidney transplant recipient.
6. The method of claim 1 , wherein the blood sample in (a) comprises whole blood, peripheral blood, serum, plasma, peripheral blood lymphocytes (PBLs), peripheral blood mononuclear cells (PBMCs), T cells, CD4 T cells, CD8 T cells, or macrophages.
7. The method of claim 1 , comprising administering the new immunosuppressive drug to the transplant recipient that has the serum creatinine level that is stable and less than 2.3 mg/dL based on the expression levels detected in (b).
8. The method of claim 1 , further comprising performing a kidney biopsy or serum creatinine test to detect subAR or risk thereof based on the expression levels detected in (b).
9. The method of claim 1 , comprising performing the Next Generation sequencing assay on the nucleic acids of interest obtained in (a).
10. The method of claim 1 , comprising performing the microarray assay on the nucleic acids of interest obtained in (a) wherein the performing the microarray assay on the nucleic acids of interest obtained in (a) comprises hybridizing the mRNA or DNA complements of mRNA from the transplant recipient that has the serum creatinine level that is stable and less than 2.3 mg/dL to a probe set, wherein the probe set comprises one or more oligonucleotides and wherein the probe set specifically detects subAR in a transplant patient with stable serum creatinine levels.
11. The method of claim 1 , wherein the transplant recipient that has the serum creatinine level that is stable and less than 2.3 mg/dL has a normal or stable eGFR.
12. The method of claim 1 , wherein the transplant recipient that has the serum creatinine level that is stable and less than 2.3 mg/dL has a serum creatinine not exceeding 20% of a baseline value of the kidney transplant recipient.
13. The method of claim 1 , additionally comprising reporting the subclinical acute rejection detected in (c) or the expression levels detected in (b) to a caregiver.
14. The method of claim 1 , wherein the immunosuppressive drug or the new immunosuppressive drug is a calcineurin inhibitor, an mTOR inhibitor, an anti-proliferative, a corticosteroid, or an anti-T-cell antibody.
15. The method of claim 1 , wherein the immunosuppressive drug or new immunosuppressive drug is cyclosporine, tacrolimus, sirolimus, everolimus, azathioprine, mycophenolic acid, prednisone, hydrocortisone, basiliximab, daclizumab, Orthoclone, anti-thymocyte globulin or anti-lymphocyte globulin.
16. The method of claim 1 , further comprising repeating (a)-(c) after administration of the new immunosuppressive drug or higher dose of the immunosuppressive drug.
17. The method of claim 1 , comprising administering a new immunosuppressive drug in (d).
18. The method of claim 17 , comprising terminating administration of the new immunosuppressive drug after repeating (a)-(c).
19. The method of claim 1 , comprising detecting expression levels of at least five genes capable of specifically detecting subclinical acute rejection selected from the group consisting of the genes recited in Tables 2, 3, 4, 7, 8, 11, 12, 14, 15, 17, and 18.
20. The method of claim 1 , comprising performing the microarray assay on the nucleic acids of interest obtained in (a).