Modeling Cystic Fibrosis Using Pluripotent Stem Cell-Derived Human Pancreatic Ductal Epithelial Cells

Simsek, Senem; Zhou, Ting; Robinson, Christopher L.; Tsai, Su-Yi; Crespo, María Eugenia Olivares; Amin, Sadaf; Lin, Xiangyi; Hon, Jane Date; Evans, Todd; Chen, Shuibing

doi:10.5966/sctm.2015-0276

Cited by 53 publications

(38 citation statements)

References 28 publications

(40 reference statements)

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“…These are currently being explored for cell replacement therapy, as these cells can spontaneously differentiate into SC-β cells after transplantation over the course of months [3,[27][28][29], but the mechanism of this maturation process is unknown and differs based on rodent species [27]. SC-β cells or earlier progenitors, particularly derived from diabetic patients through an induced pluripotent stem cell (iPSC) intermediate, are currently being studied for disease modeling and drug screening purposes [3,[30][31][32][33][34][35][36][37][38][39][40][41]. These studies would benefit from a cellular assembly that more closely reflects the native islet microenvironment [42].…”

Section: Discussionmentioning

confidence: 99%

Hydrogel platform for in vitro three-dimensional assembly of human stem cell-derived β cells and endothelial cells

Augsornworawat¹,

Velazco-Cruz²,

Song³

et al. 2019

Preprint

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Differentiation of stem cells into functional replacement cells and tissues is a major goal of the regenerative medicine field. However, one limitation has been organization of differentiated cells into multi-cellular, three-dimensional assemblies. The islets of Langerhans contain many endocrine and non-endocrine cell types, such as insulin-producing β cells and endothelial cells. Transplantation of exogenous islets into diabetic patients can serve as a cell replacement therapy, replacing the need for patients to inject themselves with insulin, but the number of available islets from cadaveric donors is low. We have developed a strategy of assembling human embryonic stem cell-derived β cells with endothelial cells into three-dimensional aggregates on a hydrogel. The resulting islet organoids express β cell markers and are functional, capable of undergoing glucose-stimulated insulin secretion. These results provide a platform for evaluating the effects of the islet tissue microenvironment on human embryonic stem cell-derived β cells and other islet endocrine cells to develop tissue engineered islets.

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Section: Discussionmentioning

confidence: 99%

Hydrogel platform for in vitro three-dimensional assembly of human stem cell-derived β cells and endothelial cells

Augsornworawat¹,

Velazco-Cruz²,

Song³

et al. 2019

Preprint

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“…In fact, as iterated in the introduction, the use of iPSCs to reconstitute [wildtype]CFTR in CF cholangiocytes has been reported . Similarly, iPSCs have also been developed to reconstitute [wildtype]CFTR expression in pancreatic ductal cells . In addition, inflammation in the pancreas might still have consequences for lung disease, independent of mutant CFTR.…”

Section: What Symptoms Must Be Suppressed By a Durable Therapy In Thementioning

confidence: 99%

“…15,16 Similarly, iPSCs have also been developed to reconstitute [wildtype]CFTR expression in pancreatic ductal cells. 14 In addition, inflammation in the pancreas might still have consequences for lung disease, independent of mutant CFTR.…”

Section: The Bystander/modifier Gene Problemmentioning

confidence: 99%

“…CFTR is a cAMP‐activated chloride channel, which, when properly glycosylated, trafficks to the apical plasma membrane of epithelial cells as a higher molecular weight “C” band. Importantly, equivalent experiments have been done by other investigators as well, not only for lung, but also for CF pancreatic duct and CF cholangiocytes . Thus for treating CF lung disease with iPSCs, the vision for this workflow would be as follows: (i) to obtain fibroblasts from a CF patient; (ii) to induce iPSCs from the fibroblasts; (iii) to correct the CFTR mutation, either with CRISPR/Cas9 or zinc‐finger nucleases (ZFNs); (iv) to grow up unlimited numbers of corrected iPSCs; and (v) to induce iPSCs to differentiate; and (vi) to deploy them back into the CF patient who donated the original fibroblast as personalized medicine.…”

Section: Introduction: the Ipsc Promise For Repairing The Cystic Fibrmentioning

confidence: 99%

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Induced pluripotent stem cells for treating cystic fibrosis: State of the science

Pollard¹,

Pollard

2018

Pediatric Pulmonology

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Induced pluripotent stem cells (iPSCs) are a recently developed technology in which fully differentiated cells such as fibroblasts from individual CF patients can be repaired with [wildtype] CFTR, and reprogrammed to differentiate into fully differentiated cells characteristic of the proximal and distal airways. Here, we review properties of different epithelial cells in the airway, and the in vitro genetic roadmap which iPSCs follow as they are step-wise differentiated into either basal stem cells, for the proximal airway, or into Type II Alveolar cells for the distal airways. The central theme is that iPSC-derived basal stem cells, are penultimately dependent on NOTCH signaling for differentiation into club cells, goblet cells, ciliated cells, and neuroendocrine cells. Furthermore, given the proper matrix, these cellular progenies are also able to self-assemble into a fully functional pseudostratified squamous proximal airway epithelium. By contrast, club cells are reserve stem cells which are able to either differentiate into goblet or ciliated cells, but also to de-differentiate into basal stem cells. Variant club cells, located at the transition between airway and alveoli, may also be responsible for differentiation into Type II Alveolar cells, which then differentiate into Type I Alveolar cells for gas exchange in the distal airway. Using gene editing, the mutant CFTR gene in iPSCs from CF patients can be repaired, and fully functional epithelial cells can thus be generated through directed differentiation. However, there is a limitation in that the lung has other CFTR-dependent cells besides epithelial cells. Another limitation is that there are CFTR-dependent cells in other organs which would continue to contribute to CF disease. Furthermore, there are also bystander or modifier genes which affect disease outcome, not only in the lung, but specifically in other CF-affected organs. Finally, we discuss future personalized applications of the iPSC technology, many of which have already survived the "proof-of-principle" test. These include (i) patient-derived iPSCs used as a "lung-on-a-chip" tool for personalized drug discovery; (ii) replacement of mutant lung cells by wildtype lung cells in the living lung; and (iii) development of bio-artificial lungs. It is hoped that this review will give the reader a roadmap through the most complicated of the obstacles, and foster a guardedly optimistic view of how some of the remaining obstacles might one day be overcome.

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“…With the current progress of research in this field, SC-β cells could be used a cell source in the near future, which meet the current need for cell sourcing, and a multi-faceted approach to analyzing the function of these cells that includes glucose-stimulated insulin secretion, mitochondrial respiration, and cytoplasmic calcium measurements with a microfluidic 11 approach could be key to properly characterizing these cells. This would synergize with other efforts in the field to study SC-β cells or earlier progenitors for disease modeling purposes, particularly with patient-derived cells [30][31][32][33][34][35][36][37][38][39][40][41][42] .…”

mentioning

confidence: 94%

Assessment of the in vitro function of human stem cell-derived β cells

Srivatsava¹,

Shahan²,

Gutgesell³

et al. 2019

Preprint

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Insulin-producing human embryonic stem cell-derived β (SC-β) cells are a promising cell source for diabetes cell replacement therapy. We have recently reported a differentiation strategy that produces SC-β cells in islet organoids that not only undergo glucose-stimulated insulin secretion but also have an islet-like dynamic insulin release profile, displaying both first and second phase insulin secretion. The goal of this study was to further characterize the functional profile of these SC-β cells in vitro. We utilized a Seahorse extracellular flux analyzer to measure mitochondrial respiration of SC-β cells at low and high glucose. We also used photolithography to fabricate a microfluidic device containing microwells to immobilize SC-β cells for perfusional analysis, monitoring cytoplasmic calcium using Fluo-4 AM at low and high glucose. Here we find that in addition to increased insulin secretion, SC-β cells have increased cellular respiration and cytoplasmic calcium ion concentration in response to a high glucose stimulation. Our results indicate that SC-β cells have similar function to that reported for islets, providing further performance characterization that could help with eventual development for diabetes cell therapy and drug screening.3 Diabetes Mellitus (DM) is a group of metabolic disorders that leads to the inability of the body to regulate blood glucose levels. Type 1 diabetes (T1D) involves the autoimmunemediated destruction of the insulin-producing pancreatic β cells located in the islets of Langerhans, leading to insulin deficiency and hyperglycemia. T1D is typically managed by injection of exogenous insulin. However, this clinical intervention does not emulate the normal behavior of the native β cells, making patients at risk for many long term complications 1 . An alternative therapy for T1D is the transplantation of cadaveric human islets, with the hope of reducing hyper-and hypoglycemic episodes. Some patients who were transplanted with islets have remained insulin independent for several years 2 . A major limitation of this approach, however, is the scarcity and quality of islets sourced from cadavers, limiting the widespread application of this therapy. Differentiation of human embryonic stem cells (hESCs) to insulinproducing β (SC-β) cells in islet organoids could serve as an unlimited supply of cells to treat millions of patients 3 , particularly if combined with transplantation strategies that vascularization, allow retrievability, and/or protect the cells from immune attack 4-8 .We recently reported a strategy for making large numbers of SC-β cells from hESCs 9 . This protocol is highly efficient, generating an almost pure population of pancreatic endocrine, of which most cells express insulin. These cells are highly functional, capable of undergoing glucose-stimulated insulin secretion. These cells were capable of restoring glucose tolerance when transplanted into streptozotocin-treated mice. Of particular note was the ability of these cells to display first and second phase insulin s...

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Modeling Cystic Fibrosis Using Pluripotent Stem Cell-Derived Human Pancreatic Ductal Epithelial Cells

Cited by 53 publications

References 28 publications

Hydrogel platform for in vitro three-dimensional assembly of human stem cell-derived β cells and endothelial cells

Hydrogel platform for in vitro three-dimensional assembly of human stem cell-derived β cells and endothelial cells

Induced pluripotent stem cells for treating cystic fibrosis: State of the science

Assessment of the in vitro function of human stem cell-derived β cells

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