Document text
Principal Investigator: Anna R. Smith
Organization: BOSTON UNIVERSITY MEDICAL CAMPUS
Fiscal Year: 2023
Award: $47,694
Funding agency: National Institute of Diabetes and Digestive and Kidney Diseases
ABSTRACT
Alpha-1 antitrypsin deficiency (AATD) is a genetic disease most commonly caused by the Z point mutation in
the SERPINA1 gene, resulting in misfolded Z-AAT protein polymerization in hepatocytes, cell death, and often
cirrhosis. Replacement of ZZ with wild type MM hepatocytes via liver transplantation is the only available cure
for AATD liver disease. However, organs for transplant are in short supply and transplantation comes with risk
of graft rejection. Hepatocyte-like cells (HLCs), derived from AATD patient induced pluripotent stem cells (iPSCs)
after gene editing of the Z mutation, could provide an unlimited supply of autologous M-AAT producing cells for
transplantation without the burden of rejection. Yet, poor engraftment of HLCs remains a critical gap that
must be addressed before iPSC-based therapy can be made available to AATD patients suffering from
severe liver disease.
The goal of this proposal is to fill this gap by promoting iPSC-derived HLC survival, proliferation and
maturation, key features for cell engraftment, to treat the AATD associated liver disease of the NSG-PiZ
transgenic mouse model. To do so, we will employ AATD patient-specific iPSCs that have been gene edited
from ZZ to MM. These MM iPSCs will be engineered to make MM HLCs that express physiological levels of the
2 key known hepatocyte mitogen receptors, hepatocyte growth factor (HGF) receptor, cMET, and epidermal
growth factor (EGF) receptor, EGFR, as well as 3 transcription factors known to be critical for hepatocyte
maturation, ATF5, PROX1, and CEBPA, using a timely controlled doxycycline inducible piggyBac transposon
system. The receptors, cMET and EGFR, will be activated using the corresponding ligands, HGF and EGF,
delivered via intravenous injection of nucleoside-modified mRNA encapsulated in lipid nanoparticles (mRNA-
LNP), a non-integrative and safe technology that our lab has recently established to treat various liver diseases.
Our preliminary data support the feasibility of this project and are summarized as follows: (1) We have built the
transcriptional units of the doxycycline-inducible piggyBac platform, and we expect to complete the platform and
start engineering hiPSC lines when the award will be initiated; (2) We showed that diseased hepatocytes in NSG-
PiZ mice are efficiently transfected with mRNA-LNPs, validating the mRNA-LNP tool to deliver mitogens in the
liver of these mice; (3) We showed that HGF+EGF mRNA-LNP treatment enhances transplanted control primary
human hepatocyte engraftment and also improves, albeit transiently, HLC survival after transplantation into
NSG-PiZ mice. This leaves room for improvement, the goal of the present application. Thus, our central
hypothesis is: Activation of the mitogen HGF/cMET and EGF/EGFR axes in combination with expression of 3
key hepatocyte maturation factors ATF5, PROX1, and CEBPA will lead to successful HLC therapy for AATD
liver disease. Importantly, this project will pioneer the use of mRNA-LNPs, which have been widely validated as
safe with the recent mRNA-based vaccines, to harness HLC-based liver therapy for AATD patients.
Terms: <2019-nCoV vaccine><A1PI><AAT deficiency><Address><Anthelone U><Antiproteases><Autologous><Award><BMP4><Basal Transcription Factor><Basal transcription factor genes><Blood><Blood Reticuloendothelial System><Blood Serum><C/EBP-alpha><CEBPA><CEBPA gene><COVID-19 vaccine><COVID19 vaccine><Cell Body><Cell Culture Techniques><Cell Cycle><Cell Death><Cell Division Cycle><Cell Line><Cell Maturation><Cell Survival><Cell Therapy><Cell Transplantation><Cell Viability><CellLine><Cells><Cirrhosis><Clinical><Culture Media><Data><Disease><Disease model><Disorder><Doxycycline><E2A Immunoglobulin Enhancer Binding Factor E12><E2A Immunoglobulin Enhancer Binding Factor E47><EGF Receptor><EGFR><ERBB Protein><Emphysema><Encapsulated><Endopeptidase Inhibitors><Engineering><Engraftment><Epidermal Growth Factor><Epidermal Growth Factor Receptor><Epidermal Growth Factor Receptor Kinase><Epidermal Growth Factor Receptor Protein-Tyrosine Kinase><Epidermal Growth Factor-Urogastrone><Epidermal Growth Factor-Urogastrone Receptors><Esteroproteases><Gene Transcription><General Transcription Factor Gene><General Transcription Factors><Generalized Growth><Genes><Genetic Alteration><Genetic Change><Genetic Diseases><Genetic Transcription><Genetic defect><Goals><Graft Rejection><Grafting Procedure><Growth><Growth Agents><Growth Factor><Growth Factor Receptors><Growth Substances><HER1><HGF Receptor><HGF gene><HGF/SF><HGFR><Hepatic><Hepatic Cells><Hepatic Cirrhosis><Hepatic Disorder><Hepatic Parenchymal Cell><Hepatic Transplantation><Hepatocyte><Hepatocyte Growth Factor><Hepatocyte Growth Factor Receptor><Hepatocyte transplant><Hepatocyte transplantation><Hepatopoietin A><Human><Human Engineering><Immune><Immunes><Immunoglobulin Transcription Factor 1><In Vitro><Injections><Kappa-E2-Binding Factor><Ligands><Liver><Liver Cells><Liver Cirrhosis><Liver Grafting><Liver Transplant><Liver diseases><Lung><Lung Fibroblast-Derived Mitogen><Lung Respiratory System><MET Protooncogene><MET gene><Messenger RNA><Mice><Mice Mammals><Mitogen Receptors><Mitogens><Modern Man><Murine><Mus><Mutation><Nucleosides><Organ Transplantation><Organ Transplants><Pathway interactions><Patients><Peptidase Inhibitors><Peptidases><Peptide Hydrolase Inhibitors><Peptide Hydrolases><Peptide Peptidohydrolase Inhibitors><Physiologic><Physiological><Point Mutation><Polymers><Production><Proliferating><Protease Antagonists><Protease Gene><Protease Inhibitor><Proteases><Proteinase Inhibitors><Proteinases><Proteins><Proteins Growth Factors><Proteolytic Enzymes><Pulmonary Emphysema><RNA Expression><RNA vaccine><RNA-based vaccine><Receptor Protein><Risk><SARS-CoV-2 vaccine><SARS-CoV2 vaccine><SARS-coronavirus-2 vaccine><Scatter Factor><Serum><Severe Acute Respiratory Syndrome CoV 2 vaccine><Severe acute respiratory syndrome coronavirus 2 vaccine><Specific qualifier value><Specified><Strains Cell Lines><System><TGF-alpha Receptor><Technology><Testing><Therapeutic><Tissue Growth><Toxic effect><Toxicities><Transcription><Transcription Factor 3><Transcription Factor E2-Alpha><Transcription Factor Proto-Oncogene><Transcription factor genes><Transfection><Transforming Growth Factor alpha Receptor><Transgenic Mice><Transplant Rejection><Transplantation><Transplantation Rejection><Urogastrone><Urogastrone Receptor><Vibramycin><a1-antitrypsin deficiency><alpha 1 Antiprotease><alpha 1-Antiproteinase><alpha 1-Antitrypsin><alpha 1-Antitrypsin Deficiency><alpha 1-Antitrypsin Trypsin Inhibitor><alpha 1-Protease Inhibitor><alpha 1-Proteinase Inhibitor><alpha-1-anti-trypsin deficiency><alpha-6-Deoxyoxytetracycline><alpha1-antitrypsin deficiency><alternative treatment><beta-Urogastrone><c-erbB-1><c-erbB-1 Protein><cell culture><cell cultures><cell engineering><cell mediated therapies><cell-based therapeutic><cell-based therapy><cellular engineering><cellular therapeutic><cellular therapy><cirrhotic><corona virus disease 2019 vaccine><coronavirus disease 2019 vaccine><coronavirus disease-19 vaccine><cultured cell line><directed differentiation><disorder model><emphysematous><erbB-1><erbB-1 Proto-Oncogene Protein><erbBl><genetic condition><genetic disorder><genome mutation><growth media><hepatic body system><hepatic disease><hepatic organ system><hepatocyte engraftment><hepatopathy><iPS><iPSC><iPSCs><improved><in vivo><induced pluripotent cell><induced pluripotent stem cell><inducible expression><inducible gene expression><inducible pluripotent stem cell><inhibitor><intravenous injection><lipid based nanoparticle><lipid nanoparticle><liver disorder><liver function><liver transplantation><mRNA><mRNA vaccine><mRNA-based vaccine><mouse model><murine model><nCoV vaccine><nCoV-19 vaccine><nCoV19 vaccine><necrocytosis><ontogeny><organ allograft><organ graft><organ xenograft><pathway><polymer><polymeric><polymerization><preconditioning><prevent><preventing><proto-oncogene protein c-erbB-1><pulmonary><receptor><regenerative><stem cell based therapy><stem cell mediated therapy><stem cell therapeutics><stem cell therapy><stem cell treatment><stem cell-based treatment><tool><transcription factor><transplant><vaccine against 2019-nCov><vaccine against SARS-CoV-2><vaccine against SARS-CoV2><vaccine against SARS-coronavirus-2><vaccine against Severe Acute Respiratory Syndrome CoV 2><vaccine against Severe acute respiratory syndrome coronavirus 2><vaccine for novel coronavirus><α-1 anti-trypsin deficiency><α-1-antitrypsin deficiency><α1-Antitrypsin><α1-Antitrypsin Deficiency><α1-Proteinase Inhibitor>