Tissue Engineering Models of Polycystic Kidney Disease.
Dominant Polycystic Kidney Disease (ADPKD) is a life-threatening monogenic disorder
where innumerable cysts develop in the kidney, leading to renal failure. There is no
known cure, and animal studies, while beneficial, often yield mixed results when
translated to the human condition. Thus, new opportunities present itself for the study
of human ADPKD by utilizing tissue en... read moregineering principles of disease modeling. However,
modeling cystic diseases in vitro presents a unique challenge as cyst morphogenesis, in
addition to complex intercellular interactions, is also governed by synergistic spatial,
mechanical and temporal effects. This thesis reports the development of kidney-like
tissue structures for normal and diseased (cystic) states using commercially available
human kidney cells. Gene silencing is used to simulate autosomal dominant polycystic
kidney disease, as inactivating mutations in polycystins -1 and/or -2 are responsible
for the disease in vivo. Our system utilizes extracellular-matrix molecules infused in
slow degrading porous silk scaffolds, which provides a 3D microenvironment for proper
cell polarization (ECM), while exhibiting structural robustness and tension (silk
scaffold). Our results indicate development of cyst-like structures in a 3D environment,
while also demonstrating the respective normal and altered phenotypes concurrent with
normal tissue and patient-derived ADPKD tissue. The structural and functional features
of kidney-like tissue structures were further characterized based on distribution of
E-cadherin, N-cadherin, transport phenomena of 6-carboxyfluorescein, and cell-matrix
interactions through integrin signaling. Importantly, this 3D in vitro model may be
further extended via perfusion reactor for long term studies of ADPKD or other renal
cystic diseases, and may have beneficial use as a therapeutic drug screening
Thesis (M.S.)--Tufts University, 2012.
Submitted to the Dept. of Biomedical Engineering.
Advisor: David Kaplan.
Committee: Ronald Perrone, and Qiaobing Xu.
Keyword: Biomedical engineering.read less