Abstract
The development of a technique to induce the transformation of somatic cells to a pluripotent state via the ectopic expression of defined transcription factors was a transformational event in the field of regenerative medicine. The development of this technique also impacted ophthalmology, as patient-specific induced pluripotent stemcells (iPSCs) may be useful resources for some ophthalmological diseases. The lens is a key refractive element in the eye that focuses images of the visual world onto the retina. To establish a new model for drug screening to treat lens diseases and investigating lens aging and development, we examined whether human lens epithelial cells (HLECs) could be induced into iPSCs and if lens-specific differentiation of these cells could be achieved under defined chemical conditions. We first efficiently reprogrammed HLECs from age-related cataract patients to iPSCs with OCT-4, SOX-2, and KLF-4. The resulting HLEC-derived iPS (HLE-iPS) colonies were indistinguishable from human ES cells with respect to morphology, gene expression, pluripotent marker expression and their ability to generate all embryonic germ-cell layers. Next, we performed a 3-step induction procedure: HLE-iPS cells were differentiated into large numbers of lens progenitor-like cells with defined factors (Noggin, BMP and FGF2), and we determined that these cells expressed lens-specific markers (PAX6, SOX2, SIX3, CRYAB, CRYAA, BFSP1, and MIP). In addition, HLE-iPS-derived lens cells exhibited reduced expression of epithelial mesenchymal transition (EMT) markers compared with human embryonic stem cells (hESCs) and fibroblast-derived iPSCs. Our study describes a highly efficient procedure for generating lens progenitor cells from cataract patient HLEC-derived iPSCs. These patient-derived pluripotent cells provide a valuable model for studying the developmental and molecular biological mechanisms that underlie cell determination in lens development and cataract pathophysiology.
Highlights
Age-related cataracts are one of the most prevalent ocular conditions and are responsible for nearly half of the cases of blindness worldwide [1]
Derivation of induced pluripotent stemcells (iPSCs) from human lens epithelial cells (HLECs) To identify a strategy for generating iPSCs from HLECs, the cultured HLECs from the cataract patient were infected with lentiviruses carrying DNA constructs encoding KLF-4, OCT-4 and SOX-2 to generate induced pluripotent stem cells (HLECiPSCs)
This was a 56-year-old male patient diagnosed as agerelated cataract and there is no role of traumatic, metabolic or genetic factors influencing the generation and differentiation of this patient’s iPSCs
Summary
Age-related cataracts are one of the most prevalent ocular conditions and are responsible for nearly half of the cases of blindness worldwide [1]. The pathogenesis of cataracts is complex and involves both genetic and environmental factors. In contrast to the cellular and molecular complexities of most ocular tissues, the lens is a relatively simple structure. The lens is one of the most promising tissues for aging studies, due to the ease of obtaining lens epithelial and fiber cells, as well as the relative molecular simplicity of fully differentiated fiber cells [2]. Developmental defects in the lens are major causes of blindness and visual impairment among children. Because many of the pathways required for lens formation are important for lens maintenance, a detailed understanding of lens development will provide a rational basis for the treatment of childhood cataract and may shed light on the lens-associated diseases observed during the aging process
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