Embryonic stem cells (ESCs) are considered a promising cell source for regenerative medicine. Intensive research over the past decade has led to the possibility that ESC-differentiated cells (ESC-DCs) could be used for the treatment of human diseases. However, increasing evidence indicates that ESC-DCs generated by the current differentiation methods are not fully functional. Recent studies indicate that ESC-DCs lack innate immunity to a wide range of infectious agents. When used in the patient, ESC-DCs would be placed in a wound site that is exposed to various pathogens and inflammatory cytokines;therefore, their viability and functionality could be compromised if the cells do not have competent immunity. We recently demonstrated that ESCs are intrinsically deficient in expressing type-I IFN and inflammatory cytokines. This finding explains the lack of innate immunity in ESC-DCs and has led to our central hypothesis that innate immunity is not developed in ESCs and cannot be effectively induced by current methods of differentiation, but it could be induced if proper """"""""immunostimulation"""""""" is provided during differentiation. The proposed study aims to understand the molecular mechanisms that control innate immunity and to develop differentiation strategies that generate ESC-DCs with competent immunity. Fibroblasts (FBs) are major cells that play key roles in tissue formation and in modulating tissue immune/inflammatory responses. Using ESC-differentiated FBs (ESC-FBs) as a model system, we will determine the molecular basis for innate immunity deficiency in ESCs and identify the factors that stimulate innate immunity development (aim 1). We will then develop novel strategies that utilize IFN?, viral RNA analogs, and lipopolysaccharide as """"""""immunostimulants"""""""" to promote innate immunity during differentiation (aim 2). The innate immunity of ESC-FBs will be comparatively analyzed with naturally differentiated FBs by in vitro models and will be evaluated with a syngeneic mouse in vivo model, where immune and inflammatory responses in implanted ESC-FBs will be assessed after the host is challenged with bacterial and viral infections (aim 3). We expect that the results will lead to the development of differentiation strategies that could fundamentally transform the current methods and achieve a better understanding of mechanisms that control innate immunity development during embryogenesis.
Embryonic stem cells (ESCs) are considered a promising cell source for regenerative medicine, but clinically usable cells have not yet been generated. An important concern is that ESC-differentiated cells from in vitro cell culture do not have active innate immunity, which could compromise their intended function. The proposed study aims to develop strategies that can generate ESC-differentiated fibroblasts that have active immunity against bacterial and viral infection. We expect that the findings may lead to the development of novel strategies that fundamentally improve the current methods and a better understanding of mechanisms that control innate immunity development during embryogenesis.
|D'Angelo, William; Chen, Bohan; Gurung, Chandan et al. (2018) Characterization of embryonic stem cell-differentiated fibroblasts as mesenchymal stem cells with robust expansion capacity and attenuated innate immunity. Stem Cell Res Ther 9:278|
|Paul, Amber M; Acharya, Dhiraj; Neupane, Biswas et al. (2018) Congenital Zika Virus Infection in Immunocompetent Mice Causes Postnatal Growth Impediment and Neurobehavioral Deficits. Front Microbiol 9:2028|
|D'Angelo, William; Gurung, Chandan; Acharya, Dhiraj et al. (2017) The Molecular Basis for the Lack of Inflammatory Responses in Mouse Embryonic Stem Cells and Their Differentiated Cells. J Immunol 198:2147-2155|
|Guo, Yan-Lin (2017) Utilization of different anti-viral mechanisms by mammalian embryonic stem cells and differentiated cells. Immunol Cell Biol 95:17-23|
|D'Angelo, William; Acharya, Dhiraj; Wang, Ruoxing et al. (2016) Development of Antiviral Innate Immunity During In Vitro Differentiation of Mouse Embryonic Stem Cells. Stem Cells Dev 25:648-59|
|Guo, Yan-Lin; Carmichael, Gordon G; Wang, Ruoxing et al. (2015) Attenuated Innate Immunity in Embryonic Stem Cells and Its Implications in Developmental Biology and Regenerative Medicine. Stem Cells 33:3165-73|
|Wang, Ruoxing; Wang, Jundi; Acharya, Dhiraj et al. (2014) Antiviral responses in mouse embryonic stem cells: differential development of cellular mechanisms in type I interferon production and response. J Biol Chem 289:25186-98|