Document text
Principal Investigator: VALERIE B GOUON-EVANS
Organization: BOSTON MEDICAL CENTER
Fiscal Year: 2023
Award: $536,021
Funding agency: National Institute of Diabetes and Digestive and Kidney Diseases
PROJECT SUMMARY
Liver transplantation is currently the only treatment for acute and chronic liver failure. Given the shortage of
donor livers, hepatocyte transplantation is considered a potential treatment for liver diseases and a bridge for
patients awaiting liver transplantation, but its application has been hampered by a limited supply of hepatocytes.
Human induced pluripotent stem cell (hiPSC) could provide an unlimited supply of patient-specific hepatocytes
for cell replacement therapy. However, generation of hiPSC-derived hepatocyte like cells (HLCs) that
repopulate a damaged liver remains a challenge, and a major gap that this application targets. Obstacles
to HLC therapy include poor cell engraftment and insufficient maturation. Thus to fill this gap, we propose a multi-
modular approach that includes (1) a bi-cell therapy composed of HLCs and supportive endothelial cells (ECs),
both derived from hiPSCs (2) engineering HLCs with novel genetic circuit technology to promote their
engraftment and maturation and (3) engineering ECs to support HLC-mediated liver repair. This approach is
based on our long-standing expertise in the directed differentiation of PSCs to HLCs having pioneered the use
of BMP4 to induce hepatic specification, an approach now utilized widely in the field. We initially observed that
ECs were always associated with mouse ESC-derived HLC clusters, and showed that ECs function as a niche
to repress Wnt and Notch signaling to promote HLC specification. Recently, we discovered that this EC
supportive function is dependent on activation of VEGFR2. Consistent with our data, it has been shown that a
VEGFA-VEGFR2 axis activates sinusoidal ECs in injured mouse livers to induce the expression of factors such
as WNT2 and HGF that trigger hepatocyte proliferation. Similarly, WNT2 and WNT9b secreted by sinusoidal
ECs were shown to induce hepatocyte proliferation following partial hepatectomy. Thus, bi-cell therapy with ECs
represents a potential strategy to overcome the obstacles of HLC therapies. In addition, we will enhance EC
supportive functions through modulation of VEGFR2 and downstream factors such as HGF, WNT2 and WNT9b.
In parallel, we will engineer HLCs to activate mitogen receptors cMET, EGFR and IL6R and express transcription
factors (ATF5, PROX1, CEBPA) that are critical for liver development, regeneration and the maturation of
reprogrammed fibroblast-derived HLCs. We will test that these pathways and TFs accelerate HLC engraftment
and maturation following bi-cell therapy. This proposal may have major clinical implications as VEGFR2,
cMET, EGFR and IL6R will be activated in vivo using the clinically safe lipid nanoparticle-complexed non-
integrative nucleoside-modified mRNA (mRNA-LNP). Our preliminary data indicate that mRNA-LNP encoding
HGF, EGF and IL6 significantly improve HLC survival after transplantation into liver injury mouse models. Our
central hypothesis is thus: a bi-cell therapy with HLCs and ECs engineered to promote the EC niche-HLC
crosstalk in combination with the unprecedented mRNA-LNP tool and genetic circuit hiPSC lines that this
application will pioneer in the liver cell therapy field will lead to successful HLC-based treatment for liver disease.
Terms: <Acceleration><Acute><Address><American><B-Cell Differentiation Factor Gene><B-Cell Stimulatory Factor 2 Gene><BMP4><BSF-2 Gene><BSF2 Gene><Basal Transcription Factor><Basal transcription factor genes><Beta Cadherin-Associated Protein><Beta-1 Catenin><Beta-2 Gene Interferon><C/EBP-alpha><CEBPA><CEBPA gene><CRISPR><CRISPR/Cas system><CUL-2><Cell Body><Cell Communication><Cell Communication and Signaling><Cell Function><Cell Interaction><Cell Line><Cell Process><Cell Signaling><Cell Survival><Cell Therapy><Cell Transplantation><Cell Viability><Cell physiology><Cell-to-Cell Interaction><CellLine><Cells><Cellular Function><Cellular Physiology><Cellular Process><Chronic><Clinical><Clustered Regularly Interspaced Short Palindromic Repeats><Co-culture><Cocultivation><Coculture><Coculture Techniques><Complex><Data><EGF><EGF Receptor><EGF gene><EGFR><ERBB Protein><Endothelial Cells><Endothelium><Engineering><Engraftment><Epidermal Growth Factor Receptor><Epidermal Growth Factor Receptor Kinase><Epidermal Growth Factor Receptor Protein-Tyrosine Kinase><Epidermal Growth Factor-Urogastrone Receptors><Exhibits><FLK1><Fibroblasts><General Transcription Factor Gene><General Transcription Factors><Generations><Genetic><Goals><HER1><HSF Gene><Hepatic><Hepatic Cells><Hepatic Disorder><Hepatic Failure><Hepatic Parenchymal Cell><Hepatic Transplantation><Hepatocyte><Hepatocyte Stimulatory Factor Gene><Hepatocyte transplant><Hepatocyte transplantation><Human><Human Engineering><Hybridoma Growth Factor Gene><IFNB2 Gene><IL-6 Gene><IL6><IL6 gene><INT1-Related Protein><INT1L1><IRP Gene><IRP Protein><In Vitro><Individual><Injections><Injury to Liver><Interleukin 6 (Interferon, Beta 2) Gene><Interleukin-6 Gene><Intracellular Communication and Signaling><KDR gene><Ligands><Liver><Liver Cells><Liver Failure><Liver Grafting><Liver Regeneration><Liver Transplant><Liver diseases><Mediating><Messenger RNA><Mice><Mice Mammals><Mitogen Receptors><Mitogens><Modeling><Modern Man><Murine><Mus><Natural regeneration><Nucleosides><Oncogene INT1-Like 1><PRO2286><Partial Hepatectomy><Pathway interactions><Patients><Proliferating><Receptor Activation><Receptor Protein><Recombinant Proteins><Recombinants><Regeneration><Regenerative capacity><Regenerative pathway><Repression><Role><Safety><Signal Transduction><Signal Transduction Systems><Signaling><Specific qualifier value><Specified><Strains Cell Lines><Subcellular Process><TGF-alpha Receptor><Technology><Testing><Therapeutic><Transcription Factor Proto-Oncogene><Transcription factor genes><Transforming Growth Factor alpha Receptor><Transplantation><United States><Urogastrone Receptor><VEGF Receptors><VEGFA><VEGFA gene><VEGFR><VEGFR-2><VEGFR2><VPF Receptor><Vascular Endothelial Cell Growth Factor Receptor><Vascular Endothelial Growth Factor A><Vascular Endothelial Growth Factor Receptor 2><Vascular Permeability Factor Receptor><Vasculotropin><WIngless-Type MMTV Integration Site Family, Member 2><WNT2><WNT2 gene><Waiting Lists><acute liver injury><anti-cancer immunotherapy><anticancer immunotherapy><beta catenin><biological signal transduction><c-erbB-1><c-erbB-1 Protein><cancer clinical trial><cancer immunotherapy><cell engineering><cell mediated therapies><cell replacement therapy><cell replacement treatment><cell-based therapeutic><cell-based therapy><cellular engineering><cellular therapeutic><cellular therapy><chronic liver injury><clinical relevance><clinical translation><clinically relevant><clinically translatable><cultured cell line><directed differentiation><end stage liver disease><end stage liver failure><erbB-1><erbB-1 Proto-Oncogene Protein><erbBl><experiment><experimental research><experimental study><experiments><hepatic body system><hepatic cell proliferation><hepatic cellular proliferation><hepatic damage><hepatic disease><hepatic injury><hepatic organ system><hepatocyte cell proliferation><hepatocyte cellular proliferation><hepatocyte proliferation><hepatopathy><human pluripotent stem cell><iPS><iPSC><iPSCs><immune-based cancer therapies><immunotherapy for cancer><immunotherapy of cancer><improved><in vivo><in vivo engraftment><induced pluripotent cell><induced pluripotent stem cell><inducible pluripotent stem cell><injured><lipid based nanoparticle><lipid nanoparticle><liver cell proliferation><liver cellular proliferation><liver damage><liver development><liver disorder><liver injury><liver repair><liver transplantation><mRNA><mouse model><murine model><notch><notch protein><notch receptors><novel><oncology clinical trial><overexpress><overexpression><partial excision of liver><pathway><proto-oncogene protein c-erbB-1><receptor><regenerate><regeneration ability><regeneration capacity><regeneration pathway><regenerative><social role><subtotal hepatectomy><tool><transcription factor><transplant><waitlist><β-catenin>