Project Details
Description
At present there are more than 25,000 patients waiting to receive liver transplants. The number is increasing due
to an aging US population accompanied by an increasing incidence of chronic liver diseases associated with
such disorders as alcoholic liver disease, hepatitis, MAFLD and NASH. In spite of efforts to persuade people to
serve as organ donors, the demand increasingly outstrips the supply for organ transplantation. One solution to
this problem is the ability to generate human livers in animals for liver as well as hepatocyte transplantation.
Although there are numerous protocols to differentiate human embryonic stem cells (hESCs), and inducible
pluripotent stem cells (iPSCs) ex vivo to a variety of cell types, they have encountered significant challenges in
translation to the clinic. However, it is now possible to regenerate the replica of organs/cells from one species of
animal within the body of a second species. This involves the knockout (KO) of specific developmental genes in
the blastocyst of species two; and the intra-blastocyst injection of pluripotent stem cells from species one to
generate offspring that carry organs/cell types derived from that donor. The translation of this approach requires
an efficient gene-editing technology. In fact, novel TALEN/CRISPR/Cas9 technologies provide such a rapid, and
cost-effective means to generate genetically modified animals. Accordingly, we propose to employ gene-editing
technology to knockout specific genes associated with liver development in the mouse embryo. We hypothesize
that rat liver can be generated in the mouse by the injection of rat ESCs or PSCs into CRISPR-genetically
engineered murine blastocysts and transplanted back into syngeneic rats. The studies represent a first step of
interspecies development of exogenic organs for transplantation without immunosuppression. We have
designed three Specific Aims to test our central hypothesis. Specifically, we will characterize (1) intra- and
interspecies exogenic liver and endothelium derived from HHEX KO embryos; (2) the immunology and function
of interspecies exogenic liver and endothelial development derived from HHEX KO embryos; and (3) several
approaches to enhance the generation of interspecies chimeras that include humanization of morphogen ligand-
receptor interactions. The resulting exogenic rat liver and endothelium will be transplanted back into syngeneic
rats to evaluate graft survival and functionality. The generation of whole livers that are comprised primarily of rat
hepatic and endothelial cells derived from implanted rat ESCs or PSCs would represent a paradigm shift and
provide the necessary preclinical evidence for ultimately creating human livers in animals. If successful, the
proposed research would be a game-changer that could conceivably pave the way for an alternate source of
human livers for organ and/or hepatocyte transplantation that is tailored to specific patients. In addition, this
novel, albeit somewhat high-risk approach circumvents many of the problems associated with research on
xenotransplantation. The potential impact on improved health care in the U.S. and worldwide for liver diseases
is great and represents a major step towards the goal of individualized medicine.
Status | Active |
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Effective start/end date | 9/21/22 → 7/31/24 |
Funding
- National Institute of Allergy and Infectious Diseases: $751,986.00
- National Institute of Allergy and Infectious Diseases: $758,037.00
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