Isolated Organ Evaluation and Treatment

Abstract

The present invention relates to methods of evaluating and/or treating organs during isolated organ perfusion, and kits for carrying out this evaluation.

1 . A method of evaluating an organ during isolated organ perfusion, the method comprising the steps of:
 perfusing the isolated organ with a Perfusate; and   measuring the concentration in the Perfusate of one or more biomarkers including a cell rupture marker, wherein the cell rupture marker is dsDNA, mitochondrial DNA, or a ribosome.   
     
     
         2 . A method according to  claim 1 , wherein the method further comprises measuring the concentration of an additional biomarker in the perfusate wherein the biomarker is selected from the group consisting of TF, tPA, SpA, and SpD. 
     
     
         3 . A method according to  claim 1 , wherein the isolated organ is from an animal or a human being and is a lung or lungs, a kidney, a liver, a heart, a pancreas or a bowel. 
     
     
         4 . A method according to  claim 1 , wherein the isolated organ is circulatorily isolated in vivo or is circulatorily isolated ex vivo. 
     
     
         5 . A method according to  claim 1 , wherein measurement of the concentration in the Perfusate of the one or more biomarkers is conducted continuously over a predetermined time period or repeatedly at predetermined intervals. 
     
     
         6 . A method according to  claim 1 , additionally comprising the step of comparing the measured concentration in the Perfusate of the one or more biomarkers against reference values, and determining whether or not to select the organ for transplantation on the basis of the comparison. 
     
     
         7 . A method according to  claim 1 , additionally comprising the step of comparing the measured concentration in the Perfusate of the one or more biomarkers against reference values, and treating the isolated organ by adding a component to the Perfusate on the basis of the comparison. 
     
     
         8 . A method according to  claim 7 , which additionally comprises the step of monitoring the treatment effect of the component on the isolated organ by continuing to measure the concentration of the one or more biomarkers in the Perfusate. 
     
     
         9 . A method according to  claim 7 , wherein a TF suppressor molecule is added to the Perfusate when the measured concentration of TF exceeds a reference value. 
     
     
         10 . A method according to  claim 8 , wherein the TF suppressor molecule is selected from the group consisting of: HMG-CoA reductase inhibitors, cyclooxygenase (COX) inhibitors, paclitaxel, lysophosphatidylcholine, insulin, nicotinamide, nitric oxide (NO)/or soluble guanylate cyclase activator, hydroxyurea, ethyl pyruvate, dimethyl sulfoxide (DMSO), angiotensin converting enzyme (ACE) inhibitors, adiponectin, retinoic acid, all-trans retinoic acid, vitamin D3, PGJ2, PPARα agonists activators (WY14643 and eicosatetraenoic acid), liver X receptor agonists, pentroxifylline, phenolics/resveratrol derivatives, indobufen, amiodarone, metformin, elevated intracellular cAMP, and PI3K/Akt/PKB signaling, miR-19, short hairpin RNA, hairpin ribozyme, or antisense ODN. 
     
     
         11 . A method according to  claim 7 , wherein an anti-apoptotic agent is added to the Perfusate when the measured concentration of one or more cell rupture markers exceed a reference value. 
     
     
         12 . A method according to  claim 7 , wherein a fibrinolytic agent is added to the Perfusate when the measured concentration of tPA is below a reference value. 
     
     
         13 . A method according to  claim 12 , wherein the fibrinolytic agent is Streptokinase or tPA. 
     
     
         14 . A method according to  claim 1 , wherein if, after evaluation, the organ is transplanted, it is not transplanted into the body from which it came. 
     
     
         15 . A method of treating an organ during isolated organ perfusion, the method comprising the step of perfusing the isolated organ with a Perfusate, wherein the Perfusate comprises a TF suppressor molecule. 
     
     
         16 . (canceled) 
     
     
         17 . A method according to  claim 15 , wherein the TF suppressor molecule is selected from the group consisting of: HMG-CoA reductase inhibitors, cyclooxygenase (COX) inhibitors, paclitaxel, lysophosphatidylcholine, insulin, nicotinamide, nitric oxide (NO)/or soluble guanylate cyclase activator, hydroxyurea, ethyl pyruvate, dimethyl sulfoxide (DMSO), angiotensin converting enzyme (ACE) inhibitors, adiponectin, retinoic acid, all-trans retinoic acid, vitamin D3, PGJ2, PPARα agonists activators (WY14643 and eicosatetraenoic acid), liver X receptor agonists, pentroxifylline, phenolics/resveratrol derivatives, indobufen, amiodarone, metformin, elevated intracellular cAMP, and PI3K/Akt/PKB signaling, miR-19, short hairpin RNA, hairpin ribozyme, or antisense ODN. 
     
     
         18 . A kit for analysis of a Perfusate during isolated organ perfusion, the kit comprising a combination of at least two assays comprising:
 an immunologic analysis for TF, tPA, dsDNA, SpA or SpD; or   a chromogenic analysis for TF or tPA; or   a Picogreen analysis for dsDNA.