Generation and characterization of human induced pluripotent stem cells (iPSCs) from hand osteoarthritis patient-derived fibroblasts

Abstract

Knowledge and research results about hand osteoarthritis (hOA) are limited due to the lack of samples and animal models of the disease. Here, we report the generation of two induced pluripotent stem cell (iPSC)-lines from patients with radiographic hOA. Furthermore, we wondered whether these iPSC-lines carried single nucleotide polymorphisms (SNPs) within genes that have been associated with hOA. Finally, we performed chondrogenic differentiation of the iPSCs in order to prove their usefulness as cellular models of the disease. We performed a non-integrative reprogramming of dermal fibroblasts obtained from two patients with radiographic rhizarthrosis and non-erosive hOA by introducing the transcriptional factors Oct4, Sox2, Klf4 and c-Myc using Sendai virus. After reprogramming, embryonic stem cell-like colonies emerged in culture, which fulfilled all the criteria to be considered iPSCs. Both iPSC-lines carried variants associated with hOA in the four studied genes and showed differences in their chondrogenic capacity when compared with a healthy control iPSC-line. To our knowledge this is the first time that the generation of iPSC-lines from patients with rhizarthrosis and non-erosive hOA is reported. The obtained iPSC-lines might enable us to model the disease in vitro, and to deeper study both the molecular and cellular mechanisms underlying hOA. Osteoarthritis (OA) is a prevalent musculoskeletal disease that affects the joints, and it has a substantial effect on quality life 1-4. Hand OA affects predominantly the carpometacarpal joint (CMCJ), followed by the interphalangeal joints (IPJs), both proximal and distal 4. OA of the CMCJ, also known as rhizarthrosis or thumb OA, is the most common location in women over 55 years, and the severity is usually linked to handedness. Mechanical pain at the base of the thumb and the thenar eminence are the principal clinical manifestations of this pathology 5. Erosive hand OA is another form that may involve the CMCJ of the thumb as well as IPJs, in which central erosions are found in the subcondral bone 4. Although both the individual and the societal burden of hand OA are well known, knowledge and research results about its underlying cellular and molecular mechanisms are limited 3 , mainly due to the dearth of tissue samples and lack of animal models of this pathology 4,6. Cellular in vitro models are meaningful tools to shed light on the molecular mechanisms and pathways that are involved in hand OA. Primary chondrocytes, immortalized cell lines and mesenchymal stromal cells are commonly used as in vitro models of OA 6. However, they present

Insights Into the Role and Potential of Schwann Cells for Peripheral Nerve Repair From Studies of Development and Injury

“…Considerations for cellular reprogramming include the starting cell type and reprogramming factors. A commonly used somatic cell source amenable to lineage conversion are fibroblasts, which are typically obtained from human foreskin (Malik and Rao, 2013 ; Bajpai et al, 2017 ) or 3 mm skin punch biopsies of the dermis (Streckfuss-Bömeke et al, 2013 ; Castro-Viñuelas et al, 2020 ). Lineage conversion of fibroblasts is achieved by the overexpression of lineage-specifying TFs and/or by the addition of extrinsic factors such as growth factors or small molecule antagonists or agonists, which specify alternative cell identities, repress the identity of the starting cell type, and remove epigenetic barriers to alter cell state (Lewitzky and Yamanaka, 2007 ; Qin et al, 2017 ).…”

Versatility of Induced Pluripotent Stem Cells (iPSCs) for Improving the Knowledge on Musculoskeletal Diseases

“…In addition to being used for genetic diseases, patient-specific iPSCs have been also generated from patients with complex diseases such as rheumatoid arthritis (RA) [ 86 ] and OA [ 86 , 88 , 89 ]. In these studies, iPSCs were generated from patients with OA or RA in order to check whether human synoviocyte- or chondrocyte-derived iPSCs showed differences in their chondrogenic capacity.…”

Characterization of Early-Onset Finger Osteoarthritis-Like Condition Using Patient-Derived Induced Pluripotent Stem Cells

“…OA can affect any joint in the human body; however, hands, knees, or hips are most commonly affected by the disease. Hand OA usually affects the carpometacarpal joint and interphalangeal joint and occurs most commonly in females over 50 years of age [1,2].…”

“…Induction of the expression of transcription factors such as OCT4, SOX2, KLF4, and MYC converts somatic cells into a state that is similar to embryonic stem cells [5,6]. Modeling of cartilage and cartilage-related diseases such as rheumatoid arthritis and knee OA have been studied using hiPSCs [1,7,8]. Castro-Viñuelas et al have generated and characterized hiPSCs from fibroblasts derived from a patient with hand OA [1].…”

“…Modeling of cartilage and cartilage-related diseases such as rheumatoid arthritis and knee OA have been studied using hiPSCs [1,7,8]. Castro-Viñuelas et al have generated and characterized hiPSCs from fibroblasts derived from a patient with hand OA [1]. Two cell lines were generated from 52-and 76-year-old female patients.…”