Ph.D. in Biochemistry, University of Colorado Denver and Health Sciences Center, Aurora, CO - 2002-2007
University of Colorado Denver, Department of Biochemistry and Molecular Genetics-2007
University of Colorado Denver, Department of Dermatology -2007-2009
The discovery of methods for reprogramming adult somatic cells into induced Pluripotent Stem Cells (iPSCs) has raised the possibility of producing truly personalized treatment options for numerous diseases. Similar to Embryonic Stem Cells (ESCs), iPSCs can give rise to any cell type in the body and they are amenable to genetic correction by homologous recombination (HR). These ESC-like properties of iPSCs allow for the development of permanent corrective therapies for many currently incurable disorders, including inherited skin diseases, without the need to use embryonic tissues or oocytes.
The successful development of iPSC- based therapies for genetic disorders depends on four important steps. First, cells need to be isolated from a patient in a non-invasive manner. Second, these cells have to be reprogrammed into iPSCs. During reprogramming, cells acquire ESC properties and undergo "rejuvenation", as can be determined by the elongation of telomeres and the restoration of mitochondrial function. Third, the genetic defects in generated iPSCs need to be corrected by safe approaches, preferentially through HR. Fourth, these genetically corrected patient-specific pluripotent cells have to be differentiated into the cell type relevant to their disease, followed by transplantation into the same patient as an autograft. Moreover, uncorrected iPSCs may be utilized as a source of disease-relevant patient-specific cells for in vitro disease modeling and in vivo xenograft modeling, which would provide platforms for yielding new insights into disease mechanisms and drug discovery.
Employing iPSC technology, our group is focusing on the development of experimental gene-correction strategies for the treatment of inherited skin blistering diseases such as the epidermolysis bullosa (EB) subtypes and congenital ichthyoses. Toward this goal, we have optimized a feeder-independent integration-free mRNA-based approach to generate clinically relevant patient-specific iPSCs. We do not rely on commercial sources to obtain reprogramming mRNAs and synthesize these molecules ourselves. This gives us the flexibility to optimize the reprogramming protocol for a variety of cell types. We also perform the design and the production of customized Transcription Activator-Like Effector Nucleases (TALENs) to enhance HR and achieve specific gene correction in disease patient-specific iPSCs. The generation of iPSC lines from patients with genetic skin disorders provides an unlimited supply of cells for studies to better understand these diseases, and an opportunity to repair gene defects in vitro. The genetically corrected iPSCs can be differentiated into keratinocytes and dermal fibroblasts suitable for autologous transplantation and used as a therapeutic option for patients. Our group was the first to establish the methodology for differentiating iPSCs into epidermal skin stem cells and bone progenitors, and we successfully differentiate human patient-specific iPSCs into functional skin cells capable of reconstituting normal human skin in a mouse xenograft model.
In addition to gene correction studies, we investigate the applicability of iPSCs in in vivo modeling of inherited skin disease, tissue rejuvenation and wound healing using mouse xenograft models. We are also interested in defining the mechanisms behind iPSC-driven rejuvenation of somatic tissues.
In collaboration with other groups, we investigate the potential of iPSCs to produce functional cardiomyocytes, neurons and mesenchymal stem cells with the goal to establish functional in vitro and in vivo human disease models.
Bilousova, G., and D.R. Roop. (2013). Generation of functional multipotent keratinocytes from mouse induced pluripotent stem cells. Methods Mol. Biol. 961:337-50
Ikonomou, L., Hemnes, A.H., Bilousova, G., Hamid, R., Loyd, J.E., Hatzopoulos, A.K., Kotton, D.N., Majka, S. M. and E. D. Austin. (2011). Programmatic Change: Lung Disease Research in the Era of Induced Pluripotency. Am J Physiol Lung Cell Mol Physiol. Dec;301(6):L830-5.
Huebner, A.J., Zhang, L., Chen, J., Bilousova, G. and D.R. Roop (2011). Genetically Engineered Mouse Models for Keratinization Disorders: New Insights into Pathophysiology and Therapeutic Approaches. The Color Atlas of Disorders of Keratinization. H. Ogawa. Tokyo, Japan, Kyowa Kikaku, LTD: 8-16.
Bilousova, G., Chen, J., and D.R. Roop. (2011). Differentiation of mouse induced pluripotent stem cells into a multipotent keratinocyte lineage. J Invest Dermatol. Apr;131(4):857-64. Epub 2010 Dec 9 (cover story with commentary).
Bilousova, G., D.H. Jun, K. King, S. DeLanghe, W. Chick, E. Torchia, K. Chow, D. Klemm, D.R. Roop, and S.M. Majka. (2011). Osteoblasts derived from induced pluripotent stem cells form calcified structures in scaffolds both in vitro and in vivo. Stem Cells. Feb;29(2):206-16. doi: 10.1002/stem.566.
Bilousova, G. and D.R. Roop. (2010). Altering cell fate: from thymus epithelium to skin stem cells. Cell Stem Cell 7: 419-20.
Bilousova, G., Marusyk, A., Porter, CC, Cardiff, R.D. and J. DeGregori. (2005). Impaired DNA replication within progenitor cell pools promotes leukemogenesis. PLoS Biol. 3(12): p e401, 2135-2147.