JPWO2021021612A5 - - Google Patents

Claims (20)

A method of producing hepatocytes, said method comprising:
manipulating hepatocyte-generated cells ex vivo by contacting said cells with at least one agent that promotes proliferation, regeneration, survival and/or engraftment;
transplanting the ex vivo engineered cells into an in vivo bioreactor under conditions suitable for engraftment;
and maintaining said in vivo bioreactor under conditions suitable for growing said engrafted cells and producing hepatocytes. 2. The method of claim 1 , further comprising isolating said expanded hepatocytes. 3. The method of claim 1 or 2 , wherein the produced hepatocytes are human hepatocytes. The method of any one of claims 1-3, wherein said hepatocyte-producing cells comprise primary human hepatocytes. The method of any one of claims 1-4 , wherein said at least one agent comprises an agonist that specifically binds to a growth factor receptor. 6. The method of Claim 5 , wherein said agonist comprises a small molecule or an antibody. 7. The method of claim 5 or 6 , wherein said growth factor receptor is c-MET and/or EGFR. The method of any one of claims 1-7 , wherein said at least one agent comprises a c-MET agonistic antibody and/or an EGFR agonistic antibody. The method of any one of claims 1-8, wherein the in vivo bioreactor comprises intrinsic liver injury. The method of any one of claims 1-9, wherein the in vivo bioreactor is immunosuppressed. 11. Any one of claims 1-10 , wherein said in vivo bioreactor is a mouse, rat, or pig comprising FAH deficiency, IL-2Rγ deficiency, RAG1 deficiency, RAG2 deficiency, or any combination thereof. the method of. 12. The method of claim 11 , wherein said in vivo bioreactor is a rodent or pig comprising FAH, RAG1 and/or RAG2, and IL-2Rγ deficiency (FRG). Hepatocytes produced by the method according to any one of claims 1-12. 14. A pharmaceutical composition for use in a method of treating liver disease in a subject, said pharmaceutical composition comprising the hepatocytes of claim 13, said method comprising treating said expanded hepatocytes in said subject A pharmaceutical composition comprising administering to 15. The pharmaceutical composition of claim 14, wherein said expanded hepatocytes have undergone further ex vivo manipulation prior to administration. Acute exacerbation of chronic liver failure (ACLF); drug or addiction-induced liver failure; congenital metabolic liver disease; Crigler-Najjar syndrome type 1; familial hypercholesterolemia; factor VII deficiency; Factor VIII deficiency (hemophilia A); phenylketonuria (PKU); glycogen storage disease type I; infantile Refsum disease; progressive familial intrahepatic cholestasis type 2; acute drug-induced liver failure; viral-induced acute liver failure; idiopathic acute liver failure; acute liver failure due to mushroom intoxication; postoperative acute liver failure; chronic liver disease, including alcoholic liver failure, hepatic encephalopathy, cirrhosis; and/or acute exacerbation of chronic liver failure due to alcohol use, drug use, and/or relapse of hepatitis B. , a pharmaceutical composition according to claim 14 or 15 . The pharmaceutical composition according to any one of claims 14-16 , wherein said liver disease is a genetic disorder. The pharmaceutical composition according to any one of claims 14-17 , wherein said liver disease comprises liver failure. The pharmaceutical composition according to any one of claims 14-18 , wherein said liver disease comprises a liver-associated enzyme deficiency. The pharmaceutical composition according to any one of claims 14-19 , wherein said liver disease is hereditary tyrosinemia.