Generation and differentiation of induced pluripotent stem cells reveal ankylosing spondylitis risk gene expression in bone progenitors

Abstract

Axial spondyloarthritis (axSpA), which encompasses ankylosing spondylitis, is a complex genetic disease. Aberrant bone formation is a key feature of pathogenesis that can lead to ankylosis of the spine. Our objective is to determine, whether genes whose variants confer susceptibility to AS are expressed in bone progenitors like mesenchymal stem cells (MSCs). Since MSCs from bone marrow is difficult to obtain, we first examined, whether MSCs can be derived from induced pluripotent stem cells (iPSCs). Dermal fibroblasts of two axSpA patients and one healthy control were reprogrammed into iPSCs using a Sendai virus vector encoding pluripotency genes. Pluripotency of iPSCs was examined by embryoid body formation and by testing for stem cell specific gene and protein expression using RT-PCR and immuno fluorescence. iPSCs were differentiated into MSCs by a TGFß inhibitor. MSCs were characterized by flow cytometry using lineage specific antibodies and by their capacity to develop into chondrocytes, adipocytes, and osteoblasts in lineage-specific medium. RNA-seq was applied to determine genome-wide gene expression patterns in MSCs, iPSCs, and blood. We show for the first time, that expression levels of several AS susceptibility genes (EDIL3, ANO6, HAPLN1, ANTXR2) involved in bone formation are significantly elevated in MSCs (2–15-fold; p ≤ 0.05) compared to blood or iPSCs and demonstrate that iPSC-derived MSCs can be differentiated into osteoblasts, chondrocytes, and adipocytes. We conclude, MSCs generated from patient fibroblast-derived iPSC lines are useful tools for studying functional genomics of risk genes associated with bone formation in AS pathogenesis.Electronic supplementary materialThe online version of this article (doi:10.1007/s10067-016-3469-5) contains supplementary material, which is available to authorized users.

Enhanced osteogenic differentiation of mesenchymal stem cells in ankylosing spondylitis: a study based on a three-dimensional biomimetic environment

“…Recently, our research, as well as that of other researchers, demonstrated that MSCs from AS patients (AS-MSCs) outperformed MSCs from healthy donors (HD-MSCs) in osteogenic differentiation in two-dimensional (2D) culture, which could be an important mechanism of pathological osteogenesis in AS 11,12 . However, whether AS-MSCs exhibit an enhanced osteogenic differentiation ability in vivo is still unclear.…”

Exploration of Alternative Splicing Events in Mesenchymal Stem Cells from Human Induced Pluripotent Stem Cells

“…Beyond direct iMSC-mediated effects, iMSCs have been demonstrated to have therapeutic paracrine effects, thus resulting in an attenuation of cardiomyopathy [ 12 ] and rejuvenation in aging [ 13 ]. In another application, disease-specific iMSCs have provided unparalleled opportunities as human cell-based disease models for pathological research, drug screening, and toxicity testing to discover novel therapeutic strategies for treating a variety of mesenchymal diseases, such as osteogenesis imperfecta [ 14 ], Hutchinson–Gilford progeria syndrome [ 15 ], axial spondyloarthritis [ 16 ], and fibrous dysplasia [ 17 ].…”

Correlation between C-Reactive Protein to Albumin Ratio and Disease Activity in Patients with Axial Spondyloarthritis

“…Many studies have shown that the whole blood components including neutrophils, lymphocytes, monocytes, and platelets were correlated with systemic inflammation [8][9][10]. Neutrophil to lymphocyte ratio (NLR), monocyte to lymphocyte ratio (MLR), and platelet to lymphocyte ratio (PLR) have been identified as new inflammatory markers and reported to be higher in axSpA patients and related to disease activity [11][12][13].…”