Methods to detect, treat and prevent acute cellular rejection in kidney allografts
Methods for prevention and treatment of kidney transplant rejection are described that involve determination, analysis and computation of a 3-gene molecular signature of levels of specific RNAs (IP-10 mRNA, CD3ε mRNA, and 18S rRNA) in urinary sample cells. The methods and devices described herein are diagnostic and prognostic of acute cellular rejection in kidney allografts.
1. A method comprising treating a developing or existing dysfunction or rejection of a kidney transplant in a subject when a urinary RNA sample has a diagnostic signature greater than −1.21, and the signature is determined by a method comprising:
(a) determining an absolute urinary CD3ε mRNA, IP-10 mRNA, and 18S rRNA copy numbers per microgram of total RNA in the urine RNA sample using at least one probe or primer with a SEQ ID NO: 4, 5, 6, 13, 14, 15, 28, 29, or 30 sequence; and
(b) ascertaining a diagnostic signature of developing or existing dysfunction or rejection of a kidney transplant in the subject with the following algorithm:
signature=−6.1487+0.8534 log 10 ( CD 3ε/18 S )+0.6376 log 10 ( IP -10/18 S )+1.6464 log 10 (18 S )
where:
CD3ε is an absolute urinary CD3ε mRNA copy number per microgram of total RNA in the urine sample;
IP-10 is an absolute urinary IP-10 mRNA copy number per microgram of total RNA in the urine sample; and
18S is an absolute urinary 18S rRNA copy number per microgram of total RNA in the urine sample times 10 −6 ;
to thereby detect and treat a developing or existing dysfunction or rejection of a kidney transplant in the subject.
2. The method of claim 1 , comprising hybridizing the at least one probe or primer to RNA from the urine sample, wherein the at least one probe or primer can hybridize to a CD3ε mRNA, IP-10 mRNA, or 18S rRNA.
3. The method of claim 1 , comprising hybridizing the at least one probe or primer to RNA from the urine sample, wherein at least one probe or primer can hybridize to an nucleic acid sequence that has at least 70% sequence identity or complementarity to SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:3.
4. The method of claim 1 , comprising normalizing the amount or copy number of CD3ε mRNA or IP-10 mRNA in the sample against the amount or copy number of 18S rRNA in the sample.
5. The method of claim 1 , wherein the subject has an existing dysfunction or rejection of a kidney transplant when the signature is greater than −1.21.
6. The method of claim 1 , wherein the subject will develop dysfunction or rejection of a kidney transplant when the signature is greater than −1.21.
7. The method of claim 1 , comprising monitoring a subject over time for developing or existing dysfunction or rejection of a kidney transplant in a subject.
8. The method of claim 1 , comprising monitoring a subject over time and informing the subject if there is continuing rise in the value of the diagnostic signature over time.
9. The method of claim 1 , comprising informing the subject of a developing or existing dysfunction or rejection of a kidney transplant in the subject.
10. The method of claim 1 , comprising informing the subject of a developing or existing dysfunction or rejection of a kidney transplant in the subject when the signature is greater than −1.21.
11. The method of claim 1 , comprising treating a developing or existing dysfunction or rejection of a kidney transplant in the subject.
12. The method of claim 1 , comprising treating a developing or existing dysfunction or rejection of a kidney transplant in the subject, wherein treating comprises plasmapheresis, administration of an anti-rejection agent, increasing a dosage of an anti-rejection agent that the subject is receiving or any combination thereof.
13. A composition consisting essentially of at least one probe or primer with a SEQ ID NO: 4, 5, 6, 13, 14, 15, 28, 29, or 30 sequence for each of CD3ε mRNA, IP-10 mRNA, and 18S rRNA; and an optional control or reference probe or primer.
14. A device consisting essentially of at least one probe or primer with a SEQ ID NO: 4, 5, 6, 13, 14, 15, 28, 29, or 30 sequence for each of CD3ε mRNA, IP-10 mRNA, and 18S rRNA covalently linked to a solid support, and an optional control or reference probe or primer covalently linked to a solid support.
15. A kit consisting essentially of at least one probe or primer with a SEQ ID NO: 4, 5, 6, 13, 14, 15, 28, 29, or 30 sequence for each of CD3ε mRNA, IP-10 mRNA, and 18S rRNA, and instructions for use in detecting potential or existing dysfunction or rejection of a kidney transplant in a subject.
16. The kit of claim 15 , comprising a filter to collect cells from a urine sample.
17. The method of claim 1 , wherein determining absolute urinary CDR mRNA, IP-10 mRNA, and 18S rRNA copy numbers per microgram of total RNA in the urine RNA sample comprises measurement of CD3ε mRNA, mRNA, and 18S rRNA by nucleic acid amplification and determination of CD3ε mRNA, IP-10 mRNA, and 18S rRNA copy numbers per microgram of total RNA with a standard curve.