Cell source and vascularization are two challenges to successful cell-based therapy. In this proposal we present an enabling technology for the vascularization of physiologically restored parenchymal cells derived from genetically deficient induced pluripotent stem cells (iPSC). As a proof-of-principle we have restored the low-density lipoprotein receptor (LDLR) feedback endocytosis in Familial Hypercholesterolemia (FH) iPSC. We will derive restored hepatocyte-like cells (HLC) and combine them with a unique stromal and vascularization cell source isolated from adipose tissue, known as the stromal vascular fraction (SVF). The objective of this proposal is to determine the capacity of an ectopic cell-based apheresis and metabolism device to modulate circulating LDL cholesterol (LDL-C) under hypercholesterolemic conditions. We will pursue this objective through the following Specific Aims:
Aim 1 : Determine the apheresis device LDL-C metabolism function. In this Aim we will investigate our device design for LDL-C internalization and metabolism, parenchymal cell survival, device vascularization and residual pluripotent cell elimination.
Aim 2 : Determine the apheresis device capacity to modulate circulating LDL-C. In this Aim we will investigate the capacity of the device to engraft, modulate LDL-C levels and produce human circulating proteins.
Cell-based technologies could be used for removing circulating toxins or providing missing biomolecules due to genetic deficiencies. Induced pluripotent stem cells (iPSC) could provide an unparalleled autologous cell source for therapeutic applications; however they may require modification to restore physiological function. Additionally engineered tissues are challenged by poor vascularization. In this project we propose to develop a vascularized cell-based enabling technology that is functionally modifiable to the patient's requirements. As a proof-of-principle we will deliver hepatocyte-like cells (HLC) derived from low-density lipoprotein receptor (LDLR) deficient Familial Hypercholesterolemia iPSC that have receptor mediated endocytosis feedback control restored. The FH-HLC-LDLR will be combined with a unique vascularization cell source isolated from adipose tissue. The implant will function as an ectopic device for removal and metabolism of excess circulating LDL to reduce the risk of cardiovascular disease development.
| Ramakrishnan, Venkat M; Boyd, Nolan L (2018) The Adipose Stromal Vascular Fraction as a Complex Cellular Source for Tissue Engineering Applications. Tissue Eng Part B Rev 24:289-299 |
| Omer, Linda; Hudson, Elizabeth A; Zheng, Shirong et al. (2017) CRISPR Correction of a Homozygous Low-Density Lipoprotein Receptor Mutation in Familial Hypercholesterolemia Induced Pluripotent Stem Cells. Hepatol Commun 1:886-898 |
