EP-4735066-A1 - MANUFACTURING OF BIOENGINEERED RECELLULARIZED ORGANS AND METHODS OF USE THEREOF

Abstract

Provided herein are methods and compositions relating to at least partially recellularized human organs. Various methods of decellularizing non-human animal organs and recellularizing a non-human animal extracellular matrix with cell compositions. Further provided are compositions and methods for treating a liver or other disease (such as acute liver failure) using an extracorporeal bioengineered liver or other organ.

Inventors

• ROSS, JEFF J.
• FECTEAU, Christopher J.
• BARRY, JOHN J.
• RIESGRAF, Shawn
• BERGSTROM, Isaac

Assignees

• Miromatrix Medical Inc.

Dates

Publication Date

20260506

Application Date

20240628

Claims (20)

1. 1. A method of making an at least partially recellularized liver composition, the method comprising: (a) treating a non-human animal liver with an anti-viral treatment; (b) perfusion decellularizing the non-human animal liver to obtain a decellularized extracellular matrix; (c) contacting the decellularized extracellular matrix with a cell composition comprising a population of human liver cells to form an at least partially recellularized liver composition.
2. 2. The method of claim 1, wherein the anti-viral treatment comprises irradiation of the non-human animal liver with an electron beam (E-beam).
3. 3. The method of claim 2, wherein the irradiation comprises exposing the non-human animal liver to an electron beam dose that is from about 2 kGy to about 50 kGy.
4. 4. The method of claim 2, wherein the irradiation comprises exposing the non-human animal liver to an electron beam dose that is from about 5 kGy to about 25 kGy.
5. 5. The method of claim 2, wherein the irradiation comprises exposing the non-human animal liver to an electron beam dose that is from about 10 kGy to 20 kGy.
6. 6. The method of claim 1, further comprising contacting the decellularized extracellular matrix with at least one of a peroxy acid or hydrogen peroxide.
7. 7. The method of claim 1, wherein the anti-viral treatment comprises irradiation of the non-human animal liver with an electron beam (E-beam), and wherein the method further comprises contacting the decellularized extracellular matrix with at least one of a peroxy acid or hydrogen peroxide.
8. 8. The method of claim 6 or claim 7, wherein the peroxy acid comprises peroxyacetic acid, peracetic acid, peroxy carboxylic acid, derivatives, or combinations thereof.
9. 9. The method of claim 1, further comprising contacting the decellularized extracellular matrix with an additional cell composition comprising a population of human vascular endothelial cells (HUVECs).
10. 10. The method of claim 1, wherein the cell composition comprises both a population of human liver cells and a population of HUVECs.
11. 11. The method of claim 1. wherein the population of human liver cells are primary human liver cells.
12. 12. The method of claim 1, wherein the population of human liver cells are in vitro- differentiated human liver cells.
13. 13. The method of claim 12, wherein the in vv/ro-differen ti ted human liver cells are differentiated from a population of embryonic stem cells, a population of induced pluripotent stem cells (iPSCs), or a population of adult stem cells.
14. 14. The method of claim 1, wherein the population of human liver cells are contacted with a protease prior to contacting the decellularized extracellular matrix with the cell composition in step (c).
15. 15. The method of claim 1, wherein the at least partially recellularized liver is characterized as having an increase in a level of ammonia clearance relative to a population of liver cells that are not engrafted onto a decellularized extracellular matrix.
16. 16. The method of claim 1, wherein the non-human animal liver is from a non-human mammal.
17. 17. The method of claim 16, wherein the non-human mammal is an ungulate.
18. 18. The method of claim 17, wherein the ungulate is a pig.
19. 19. The method of claim 1. wherein the non-human animal liver is frozen prior to the antiviral treatment.
20. 20. A method of making an at least partially recellularized liver composition, the method comprising: (a) treating a non-human animal liver with an anti-viral treatment; (b) perfusion decellularizing the non-human animal liver to obtain a decellularized extracellular matrix; (c) contacting the decellularized extracellular matrix with a first cell composition comprising a population of human vascular endothelial cells; and (d) contacting the decellularized extracellular matrix with a second cell composition comprising a population of human liver cells to form an at least partially recellularized liver composition.

Description

MANUFACTURING OF BIOENGINEERED RECELLULARIZED ORGANS AND METHODS OF USE THEREOF CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims priority to, and the benefit of, U.S. Provisional Application No. 63/511,413, filed June 30, 2023, the contents of which is entirely incorporated herein by reference for all purposes and commonly owned. BACKGROUND [0002] Organ transplants are common across the United States and throughout the world. Organs such as livers, kidneys, pancreas, lungs and hearts are commonly transplanted to prolong the life of the recipients. However, there is often a shortage of organs, creating a demand and a waiting list for the organs. As one example, Acute Liver Failure (ALF), can be a life-threatening, critical illness in patients that is characterized by the rapid onset of abnormal liver biochemistry, coagulopathy, and often the progression to encephalopathy. While clinical outcomes for patients with ALF have steadily improved with changes in medical management, the gold standard to treat ALF remains liver transplantation, with a 1-year survival rate of 91%. Data from the Acute Liver Failure Study Group (ALFSG) Registry and the Scientific Registry of Transplant Recipients (SRTR) reported that only 64% of patients listed for transplant received a lifesaving organ. Strategies are needed to mitigate the long waitlists for organ transplants and improve patient quality of life. SUMMARY [0003] Provided herein are bioengineered, recellularized organs for use in the treatment of various diseases. In some embodiments, the bioengineered, recellularized organ comprises a liver or a liver has reduced and/or inactivated microbial particles, can be cultured for long periods of time (e.g, greater than 4 days) prior to transplantation or use in an ex-vivo blood circuit without systemic anticoagulation, and can be stored in cold storage for more than 6 hours and maintain liver function (e.g., ammonia clearance and urea production). Further provided herein are methods of manufacturing bioengineered liver tissues and methods of treating a patient with an extracorporeal liver. [0004] Provided herein are methods of making an at least partially recellularized organ composition, including but not limited to liver, kidney, lung and heart, wherein the methods comprise: (a) treating a non-human animal organ with an anti-viral treatment; (b) perfusion decellularizing the non-human animal organ to obtain a decellularized extracellular matrix; (c) contacting the decellularized extracellular matrix with a cell composition comprising a population of human cells to form an at least partially recellularized organ composition. [0005] Provided herein are methods of making an at least partially recellularized liver composition, wherein the methods comprise: (a) treating a non-human animal liver with an antiviral treatment; (b) perfusion decellul arizing the non-human animal liver to obtain a decellularized extracellular matrix; (c) contacting the decellularized extracellular matrix with a cell composition comprising a population of human liver cells to form an at least partially recellularized liver composition. [0006] Provided herein are methods of making an at least partially recellularized organ composition, wherein the methods comprise: (a) treating a non-human animal organ with an antiviral treatment; (b) perfusion decellul arizing the non-human animal organ to obtain a decellularized extracellular matrix; (c) contacting the decellularized extracellular matrix with a cell composition comprising a population of human cells to form an at least partially recellularized organ composition. [0007] Provided herein are methods of making an at least partially recellularized organ composition, the methods comprising: (a) treating a non-human animal organ with an anti-viral treatment; (b) perfusion decellularizing the non-human animal organ to obtain a decellularized extracellular matrix; (c) contacting the decellularized extracellular matrix with a first cell composition comprising a population of human vascular endothelial cells; and (d) contacting the decellularized extracellular matrix with a second cell composition to form an at least partially recellularized organ composition. [0008] Provided herein are methods of making an at least partially recellularized liver composition, the methods comprising: (a) treating a non-human animal liver with an anti-viral treatment; (b) perfusion decellularizing the non-human animal liver to obtain a decellularized extracellular matrix; (c) contacting the decellularized extracellular matrix with a first cell composition comprising a population of human vascular endothelial cells; and (d) contacting the decellularized extracellular matrix with a second cell composition comprising a population of human liver cells to form an at least partially recellularized liver composition. [0009] Provided herein are compositions comprising an at least partially recellularized organ produc