Biomarkers related to organ function
Disclosed herein are methods of identifying biomarkers (such as genes (e.g., RNA or mRNA), proteins, and/or small molecules) that can be used to predict organ or tissue function or dysfunction. In some embodiments, the methods include ex vivo perfusion of the organ or tissue, collection of samples from the organ or tissue (for example, perfusate, fluids produced by the organ (such as bile or urine), or tissue biopsies) and measuring the level of one or more biomarkers in the sample. It is also disclosed herein that an analysis of biomarkers (such as genes (e.g., RNA or mRNA), proteins, and/or small molecules) present in a biological sample from an organ, tissue, or subject can be used to identify whether the organ, tissue, or subject is at risk for (or has) organ dysfunction or organ failure.
1. A method for identifying organ dysfunction, comprising:
obtaining a sample from an ex vivo organ undergoing machine perfusion with a perfusion solution comprising a hemoglobin-based oxygen carrier at subnormothermic temperatures, wherein the perfusion solution comprises 3-4 g/dL acellular cross-linked hemoglobin in a physiologically acceptable medium, has a pH of about 7.55-7.85 at room temperature, an osmolality of about 290-300 mOsm/kg, and a colloid osmotic pressure of about 35-65 mm Hg;
measuring the amount of glucose, lactate, pyruvate, fructose 6-phosphate, glucose 6-phosphate, valine, leucine, isoleucine, alpha-ketoglutarate, glutamate, 15-HETE, and 12-HETE in the sample;
comparing the amount of glucose, lactate, pyruvate, fructose 6-phosphate, glucose 6-phosphate, valine, leucine, isoleucine, alpha-ketoglutarate, glutamate, 15-HETE, and 12-HETE with the amount of glucose, lactate, pyruvate, fructose 6-phosphate, glucose 6-phosphate, valine, leucine, isoleucine, alpha-ketoglutarate, glutamate, 15-HETE, and 12-HETE in a reference or control from one or more organs that are known to have organ dysfunction;
determining that glucose, lactate, pyruvate, fructose 6-phosphate, glucose 6-phosphate, valine, leucine, isoleucine, alpha-ketoglutarate, glutamate, 15-HETE, and 12-HETE are biomarkers of organ dysfunction when the amount of the glucose, lactate, pyruvate, fructose 6-phosphate, glucose 6-phosphate, valine, leucine, isoleucine, alpha-ketoglutarate, glutamate, and 15-HETE is decreased compared to the reference or control and the amount of the 12-HETE is increased compared to the reference or control;
utilizing the amount of the glucose, lactate, pyruvate, fructose 6-phosphate, glucose 6-phosphate, valine, leucine, isoleucine, alpha-ketoglutarate, glutamate, 15-HETE, and 12-HETE to select the organ undergoing machine perfusion when the amount of the glucose, lactate, pyruvate, fructose 6-phosphate, glucose 6-phosphate, valine, leucine, isoleucine, alpha-ketoglutarate, glutamate, 15-HETE, and 12-HETE of is a predictor of organ function; and
transplanting the selected organ into a transplant recipient.
2. The method of claim 1 , wherein the perfusion solution comprises 3-4 g/dL cross-linked hemoglobin, 25-30 mM NaCl, 1-2 mM KCl, 17-19 mM KH 2 PO 4 , 55-65 mM sodium gluconate, 6-8 mM sodium lactate, 3-4 mM magnesium gluconate, 0.6-0.8 mM CaCl 2 dihydrate, 15-16 mM NaOH, 3-4 mM adenine, 6-8 mM dextrose, 2-3 mM glutathione, 6-8 mM HEPES, 3-4 mM ribose, 20-25 mM mannitol, 35-40 g/L hydroxyethyl starch, and 40-60 mg/dL N-acetyl-L-cysteine.
3. The method of claim 1 , wherein the sample comprises perfusion or preservation solution from the organ, a biopsy from the organ, or a fluid produced by the organ.
4. The method of claim 1 , wherein the organ is a liver, a kidney, a lung, a heart, a pancreas, a small intestine, a limb, an extremity, or a portion of any one thereof.
5. A method for identifying organ dysfunction, comprising:
obtaining a sample from an ex vivo organ undergoing machine perfusion with a perfusion solution comprising a hemoglobin-based oxygen carrier at subnormothermic temperatures, wherein the perfusion solution comprises 3-4 g/dL acellular cross-linked hemoglobin in a physiologically acceptable medium, has a pH of about 7.55-7.85 at room temperature, an osmolality of about 290-300 mOsm/kg, and a colloid osmotic pressure of about 35-65 mm Hg;
measuring the amount of glucose, lactate, pyruvate, fructose 6-phosphate, glucose 6-phosphate, valine, leucine, isoleucine, alpha-ketoglutarate, glutamate, 15-HETE, and 12-HETE in the sample;
comparing the amount of the glucose, lactate, pyruvate, fructose 6-phosphate, glucose 6-phosphate, valine, leucine, isoleucine, alpha-ketoglutarate, glutamate, 15-HETE, and 12-HETE with the amount of glucose, lactate, pyruvate, fructose 6-phosphate, glucose 6-phosphate, valine, leucine, isoleucine, alpha-ketoglutarate, glutamate, 15-HETE, and 12-HETE in a reference or control from one or more organs that are known to have organ dysfunction; and
wherein the amount of the glucose, lactate, pyruvate, fructose 6-phosphate, glucose 6-phosphate, valine, leucine, isoleucine, alpha-ketoglutarate, glutamate, and 15-HETE is decreased compared to the reference or control and the amount of the 12-HETE is increased compared to the reference or control.
6. The method of claim 5 , wherein the perfusion solution comprises 3-4 g/dL cross-linked hemoglobin, 25-30 mM NaCl, 1-2 mM KCl, 17-19 mM KH 2 PO 4 , 55-65 mM sodium gluconate, 6-8 mM sodium lactate, 3-4 mM magnesium gluconate, 0.6-0.8 mM CaCl 2 dihydrate, 15-16 mM NaOH, 3-4 mM adenine, 6-8 mM dextrose, 2-3 mM glutathione, 6-8 mM HEPES, 3-4 mM ribose, 20-25 mM mannitol, 35-40 g/L hydroxyethyl starch, and 40-60 mg/dL N-acetyl-L-cysteine.
7. The method of claim 5 , wherein the sample comprises perfusion or preservation solution from the organ, a biopsy from the organ, or a fluid produced by the organ.
8. The method of claim 5 , wherein the organ is a liver, a kidney, a lung, a heart, a pancreas, a small intestine, a limb, an extremity, or a portion of any one thereof.