Abstract

In this study, we employ in vitro models to study the factors involved in the differentiation of pancreatic precursor cells into hormone-producing cells of the islets of Langerhans and their mechanisms of action with a goal to develop a system that could be used for cell replacement therapy for patients with diabetes mellitus. Development of the endocrine pancreas includes a series of early events wherein precursor cells migrate to form aggregates that subsequently differentiate into islets of Langerhans. We use PANC-1 cells, a human pancreatic cell line that can be induced to differentiate into hormone-producing cell aggregates, and cells derived from human cadaveric pancreata, human islet-derived precursor cells (hIPCs), to study regulation of cell migration and aggregation that precede differentiation. Most interestingly, we have found that hIPCs most likely are derived by in vitro epithelial-to-mesenchymal transition of mature beta cells and that differentiation of hIPCs represents mesenchymal-to-epithelial transition back to pancreatic hormone-expressing. We have established a novel cell culture system using hIPCs that allows for proliferation of these precursor cells for at least 30 generations. The proliferating cells can be induced to differentiate into insulin-expressing cells in vitro and when transplanted in to mice increase their content of preproinsulin mRNA at least 100,000-fold. We have begun to use these cells to study their regulation by growth/developmental factors (GDFs). One set of GDFs that we have identified are the factors that are ligands for protease-activated receptors (PARs). Activation of PARs enhances the survival of hIPCs as they are induced to differentiate into hormone-expressing islet-like cell aggregates (ICAs). We are also studying GDFs that can allow for generations of hIPCs in the absence of animal serum/proteins and role of cell-cell and cell-matrix interactions that are involved in the regulation of differentiation of the three-dimensional ICA structures.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Intramural Research (Z01)
Project #
1Z01DK011007-04
Application #
7151494
Study Section
(RHA)
Project Start
Project End
Budget Start
Budget End
Support Year
4
Fiscal Year
2005
Total Cost
Indirect Cost

  Institution

Name
U.S. National Inst Diabetes/Digst/Kidney
Department
Type
DUNS #
City
State
Country
United States
Zip Code

  Publications

Hiram-Bab, Sahar; Katz, Liora S; Shapira, Hagit et al. (2014) Platelet-derived growth factor BB mimics serum-induced dispersal of pancreatic epithelial cell clusters. J Cell Physiol 229:743-51
Katz, Liora S; Geras-Raaka, Elizabeth; Gershengorn, Marvin C (2013) Reprogramming adult human dermal fibroblasts to islet-like cells by epigenetic modification coupled to transcription factor modulation. Stem Cells Dev 22:2551-60
Couty, Jean-Pierre; Lupu-Meiri, Monica; Oron, Yoram et al. (2009) Kaposi's sarcoma-associated herpesvirus-G protein-coupled receptor-expressing endothelial cells exhibit reduced migration and stimulated chemotaxis by chemokine inverse agonists. J Pharmacol Exp Ther 329:1142-7
Ikonomou, L; Geras-Raaka, E; Raaka, B M et al. (2008) Beta-catenin signalling in mesenchymal islet-derived precursor cells. Cell Prolif 41:474-91
Deshet, Naamit; Lupu-Meiri, Monica; Espinoza, Ingrid et al. (2008) Plasminogen-induced aggregation of PANC-1 cells requires conversion to plasmin and is inhibited by endogenous plasminogen activator inhibitor-1. J Cell Physiol 216:632-9
Mutskov, Vesco; Raaka, Bruce M; Felsenfeld, Gary et al. (2007) The human insulin gene displays transcriptionally active epigenetic marks in islet-derived mesenchymal precursor cells in the absence of insulin expression. Stem Cells 25:3223-33
Morton, Russell A; Geras-Raaka, Elizabeth; Wilson, Leah M et al. (2007) Endocrine precursor cells from mouse islets are not generated by epithelial-to-mesenchymal transition of mature beta cells. Mol Cell Endocrinol 270:87-93
Davani, Behrous; Ikonomou, Laertis; Raaka, Bruce M et al. (2007) Human islet-derived precursor cells are mesenchymal stromal cells that differentiate and mature to hormone-expressing cells in vivo. Stem Cells 25:3215-22
Wei, Chiju; Geras-Raaka, Elizabeth; Marcus-Samuels, Bernice et al. (2006) Trypsin and thrombin accelerate aggregation of human endocrine pancreas precursor cells. J Cell Physiol 206:322-8
Gershengorn, Marvin C; Geras-Raaka, Elizabeth; Hardikar, Anandwardhan A et al. (2005) Are better islet cell precursors generated by epithelial-to-mesenchymal transition? Cell Cycle 4:380-2

Showing the most recent 10 out of 12 publications