Liver cell transplantation has extensive potential for a variety of genetic and acquired conditions. Through pioneering work performed under this grant, we established for the first time that liver sinusoidal endothelial cells (LSEC) can be successfull transplanted. We developed suitable conditions to demonstrate engraftment of transplanted cells, including roles in this process of intrahepatic cell-cell interactions and cytokines/chemokines/receptors. Detailed analysis established that transplanted LSEC integrated in the endothelial lining of the liver and continued to express endothelial functions. Also, we established mechanisms regulating proliferation of transplanted LSEC in the liver. The ability of transplanted LSEC to synthesize and release FVIII permitted us to achieve permanent correction of hemophilia A in a mouse model. Subsequently, we established mechanisms in therapeutic gene transfer in transplanted LSEC that had been genetically-modified by lentiviral vectors. In other studies, we determined whether LSEC could originate from extrahepatic sources, e.g., bone marrow. These studies excluded that endothelial cells could originate from donor bone marrow-derived cells but we identified additional cell types, i.e., mononuclear cells, including Kupffer cells, as well as mesenchymal stromal cells derived from bone marrow with ability to express FVIII in lower levels. Finally, we identified antiviral mechanisms for hepatitisB and C replication, to consider whether disease-resistant hepatocytes and healthy endothelial cells could be simultaneously replaced. Therefore, insights in the biology of LSEC will offer suitable frameworks for cell/gene therapy in liver-related conditions. To advance cell/gene therapy, we now propose studies with human LSEC progenitor cells. Our hypothesis is that suitable endothelial progenitor cell populations can be isolated, expanded and manipulated in culture conditions from healthy donor tissues, followed by transplantation to replace liver sinusoidal endothelium, such that cell therapy in people will be advanced. As appropriate strategies are critical for inducing engraftment and proliferation in transplanted cells, we will develop studies in well-characterized and superb cell transplantation models. First, we will define properties of candidate human endothelial progenitor cells. This will be followed by examination of mechanisms to improve engraftment and proliferation in transplanted endothelial cells. These studies will permit examination of the cell therapy potential of human cells in hemophilia A mice. In this way, innovative approaches for cell therapy with human endothelial cells will overcome existing barriers in curing hemophilia and other disorders. Fundamental knowledge in respect to the properties and uses of human endothelial cells will advance insights in liver biology and liver cell therapy, which will be of enormous value for improving human health.
Liver-directed cell therapy has extensive potential for impacting human health. Already, some 100 people have undergone hepatocyte transplantation for liver failure, enzyme deficiency states, FVII deficiency, etc. (1-3). Moreover, animal studies showed that cell therapy will be helpful in other situations. Based on our work under this grant in transplantation of endothelial cells, extensive further interest has been generated in cell therapy for coagulation defects, e.g., hemophilia. Our cell transplantation studies in hemophilia mouse have shown for the first time that hemophilia can be permanently cured by cell therapy. Despite work over the past two decades, this accomplishment in hemophilia had eluded gene therapy investigators. Since hemophilia A afflicts over one million individuals worldwide, this will be highly significant for human health. Moreover, many individuals with hemophilia are burdened with associated liver diseases, e.g., chronic hepatitis B and C. Therefore, replacement of healthy LSEC and hepatocytes will eventually be appropriate for many individuals. In this way, lessons from the current proposal will be of extensive value for larger populations, since chronic viral hepatitis afflicts some 400 million people worldwide, and will have uncommon public health significance and impact in therapies for liver diseases.
|Kakabadze, Zurab; Kakabadze, Ann; Chakhunashvili, David et al. (2018) Decellularized human placenta supports hepatic tissue and allows rescue in acute liver failure. Hepatology 67:1956-1969|
|Sharma, Yogeshwar; Liu, Jinghua; Kristian, Kathleen E et al. (2018) In Atp7b-/- Mice Modeling Wilson's Disease Liver Repopulation with Bone Marrowderived Myofibroblasts or Inflammatory Cells and not Hepatocytes is Deleterious. Gene Expr :|
|Jaber, Fadi Luc; Sharma, Yogeshwar; Gupta, Sanjeev (2017) Demonstrating Potential of Cell Therapy for Wilson's Disease with the Long-Evans Cinnamon Rat Model. Methods Mol Biol 1506:161-178|
|Rogler, Charles E; Bebawee, Remon; Matarlo, Joe et al. (2017) Triple Staining Including FOXA2 Identifies Stem Cell Lineages Undergoing Hepatic and Biliary Differentiation in Cirrhotic Human Liver. J Histochem Cytochem 65:33-46|
|Merlin, Simone; Bhargava, Kuldeep K; Ranaldo, Gabriella et al. (2016) Kupffer Cell Transplantation in Mice for Elucidating Monocyte/Macrophage Biology and for Potential in Cell or Gene Therapy. Am J Pathol 186:539-51|
|Yu, Yang; Guerrero, Candace R; Liu, Shuo et al. (2016) Comprehensive Assessment of Oxidatively Induced Modifications of DNA in a Rat Model of Human Wilson's Disease. Mol Cell Proteomics 15:810-7|
|Viswanathan, Preeti; Gupta, Priya; Kapoor, Sorabh et al. (2016) Thalidomide promotes transplanted cell engraftment in the rat liver by modulating inflammation and endothelial integrity. J Hepatol 65:1171-1178|
|Zanolini, Diego; Merlin, Simone; Feola, Maria et al. (2015) Extrahepatic sources of factor VIII potentially contribute to the coagulation cascade correcting the bleeding phenotype of mice with hemophilia A. Haematologica 100:881-92|
|Dogra, Samriti; Bandi, Sriram; Viswanathan, Preeti et al. (2015) Arsenic trioxide amplifies cisplatin toxicity in human tubular cells transformed by HPV-16 E6/E7 for further therapeutic directions in renal cell carcinoma. Cancer Lett 356:953-61|
|Kumar, Mukesh; Sharma, Yogeshwar; Bandi, Sriram et al. (2015) Endogenous antiviral microRNAs determine permissiveness for hepatitis B virus replication in cultured human fetal and adult hepatocytes. J Med Virol 87:1168-83|
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