Guided tissue organization and disease modeling in a kidney tubule array

Abstract

Three-dimensional (3D) in vitro kidney tubule models have either largely relied on the self-morphogenetic properties of the mammalian cells or used an engineered microfluidic platform with a monolayer of cells cultured on an extracellular matrix (ECM) protein coated porous membrane. These systems are used to understand critical processes during kidney development and transport properties of renal tubules. However, high variability and lack of kidney tubule-relevant geometries among engineered structures limit their utility in disease research and pre-clinical drug testing. Here, we report a novel bioengineered guided kidney tubule (gKT) array system that incorporates in vivo-like physicochemical cues in 3D culture to reproducibly generate homogeneous kidney tubules. The system utilizes a unique 3D micro-molded ECM platform in human physiology-scale dimensions (50-μm diameter) and relevant shapes to guide cells towards formation of mature tubule structures. The guided kidney tubules in our array system display enhanced tubule homogeneity with in vivo-like structural and functional features as evaluated by marker protein localization and epithelial transport analysis. Furthermore, the response of gKT structures to forskolin treatment exhibits characteristic tissue transformations from tubules to expanding cysts. Moreover, acute cisplatin injury causes induction of Kidney Injury Molecule-1 (KIM-1) expression as well as tubular necrosis and apoptosis. Thus the gKT array system offers enhanced structural uniformity with accurate in vivo-like tissue architecture, and will have broad applications in kidney tubule disease pathophysiology (including ciliopathies and drug-induced acute kidney injury), and will enhance pre-clinical drug screening studies.

A Multitubular Kidney-on-Chip to Decipher Pathophysiological Mechanisms in Renal Cystic Diseases

“…No mean tubule dilation was observed with MDCK tubes. It is noteworthy to mention that MDCK tube dilation might have been triggered with some drug treatments, like cAMP agonists, as reported for renal cells in a bioengineered guided kidney tubule array system, where forskolin treatment of mIMCD3 cells induced a transformation from tubules to progressively dilating cystic structures ( Subramanian et al, 2018 ). However, the scope of our study was to study the behavior of specific ADPKD models.…”

“…While these different approaches have been focused on the study of renal transport function, morphology or collective cell organization, exploring cystic diseases with kidney-on-chips has been scarcely addressed. Recently, microlithography-based approaches were used to generate parallelepiped structures in a collagen-Matrigel matrix, with tube to cyst transition upon cAMP stimulation ( Subramanian et al, 2018 ).…”

Organ-on-a-Chip Systems for Modeling Pathological Tissue Morphogenesis Associated with Fibrosis and Cancer

“…This approach enhances the uniformity of tubule size and shape, which is advantageous for high-throughput pre-clinical drug-testing applications. Importantly, because cells are surrounded by ECM, which is permissive to cell-mediated remodeling and deformation, pathological tissue morphogenesis in response to an induced pathology can be modeled with the gKT array system . Indeed, Subramanian et al demonstrated that upon treatment with forskolin to induce acute kidney injury, cells showed disrupted apical-basal polarity and tubules formed progressively expanding cysts.…”

“…For example, the lung microtissue array uses micropillars to guide the formation of suspended 3D fibroblast-collagen microtissues into thin membranous morphologies that resemble the walls of alveolar sacs . Other organ-on-a-chip systems use internal cavities within ECM to guide the self-assembly of cells into the desired 3D morphologies (e.g., tubes). ,,, Because these organ-on-a-chip systems support tissue morphogenesis to build 3D tissues, they are well-suited for modeling pathological tissue morphogenesis.…”

Advanced bioengineering technologies for preclinical research

“…Regarding tubular structures, very recently an array of kidney tubules produced from a micro-molded platform in collagen-matrigel gels. This platform allows for the modeling of kidney diseases such as cystic kidney disease and acute kidney injury as well as drug testing purpose [104]. Finally, the small intestine crypt-villus morphology has also been reproduced by either replica molding procedures [29,105] and, more recently, by reactiondiffusion mediated photolithography ( Figure 6(c)) [106].…”