US 11,891,664 B2
Methods of detecting renal transplant rejection using microvesicle nucleic acid biomarkers
James Hurley, Marblehead, MA (US); Christine Coticchia, Waltham, MA (US); Robert Kitchen, Somerville, MA (US); Vasisht Tadigotla, Newton, MA (US); Johan Karl Olov Skog, Lincoln, MA (US); and Jamil Azzi, Boston, MA (US)
Assigned to Exosome Diagnostics, Inc., Waltham, MA (US); and The Brigham and Women's Hospital, Boston, MA (US)
Appl. No. 16/613,298
Filed by Exosome Diagnostics, Inc., Waltham, MA (US); and The Brigham and Women's Hospital, Boston, MA (US)
PCT Filed May 16, 2018, PCT No. PCT/US2018/032888
§ 371(c)(1), (2) Date Nov. 13, 2019,
PCT Pub. No. WO2018/213392, PCT Pub. Date Nov. 22, 2018.
Claims priority of provisional application 62/507,433, filed on May 17, 2017.
Prior Publication US 2020/0199675 A1, Jun. 25, 2020
Int. Cl. C12Q 1/6883 (2018.01); G01N 1/28 (2006.01); G01N 33/68 (2006.01)
CPC C12Q 1/6883 (2013.01) [C12Q 2600/158 (2013.01); G01N 1/28 (2013.01); G01N 33/6893 (2013.01); G01N 2800/245 (2013.01); G01N 2800/347 (2013.01)] 8 Claims
 
1. A method of treating a kidney transplant rejection in a patient, the method comprising the steps of:
a) isolating a microvesicle fraction from a urine sample from the patient;
b) extracting nucleic acids from the microvesicle fraction;
c) determining the expression levels of at least two biomarkers from the extracted nucleic acids, wherein the at least two biomarkers comprise each of CXCL9 and CXCL10, and the expression level of at least one normalizing gene, wherein the at least one normalizing gene comprises IL17RA;
d) normalizing the expression levels of the at least two biomarkers to the expression level of the at least one normalizing gene;
e) determining that the patient is undergoing a kidney transplant rejection based on the normalized expression levels of the at least two biomarkers; and
f) administering at least one kidney transplant rejection therapy to the patient undergoing a kidney transplant rejection.