WNT signaling determines tumorigenicity and function of ESC-derived retinal progenitors
Lu Cui, … , Guo-Tong Xu, Ying Jin
Lu Cui, … , Guo-Tong Xu, Ying Jin
Published April 1, 2013; First published March 25, 2013
Citation Information: J Clin Invest. 2013;123(4):1647-1661. https://doi.org/10.1172/JCI65048.
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Research Article

WNT signaling determines tumorigenicity and function of ESC-derived retinal progenitors

Abstract

Tumor formation constitutes a major obstacle to the clinical application of embryonic stem cell–derived (ESC-derived) cells. In an attempt to find major extracellular signaling and intrinsic factors controlling tumorigenicity and therapeutic functionality of transplanted ESC-derived retinal progenitor cells (ESC-RPCs), we evaluated multiple kinds of ESC-RPCs in a mouse retinal degeneration model and conducted genome-wide gene expression profiling. We identified canonical WNT signaling as a critical determinant for the tumorigenicity and therapeutic function of ESC-RPCs. The function of WNT signaling is primarily mediated by TCF7, which directly induces expression of Sox2 and Nestin. Inhibition of WNT signaling, overexpression of dominant-negative Tcf7, and silencing Tcf7, Sox2, or Nestin all resulted in drastically reduced tumor formation and substantially improved retinal integration and visual preservation in mice. These results demonstrate that the WNT signaling cascade plays a critical role in modulating the tumorigenicity and functionality of ESC-derived progenitors.

Authors

Lu Cui, Yuan Guan, Zepeng Qu, Jingfa Zhang, Bing Liao, Bo Ma, Jiang Qian, Dangsheng Li, Weiye Li, Guo-Tong Xu, Ying Jin

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Figure 1

ESC-RPC but not P-RPC transplantation generates ocular tumors.

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ESC-RPC but not P-RPC transplantation generates ocular tumors. (A) Immun...
(A) Immunofluorescence images of costaining of PAX6 with NESTIN, SIX3, and RAX, respectively, in ESC-RPCs at 14 days and costaining of PAX6 with OTX2 and NRL, respectively, or opsin staining alone at 24 days of the differentiation. DAPI is shown in gray. (B) Representative RT-PCR analysis of the expression for retinal markers in ESCs, P-RPCs, and ESC-RPCs. (C) FCM profiles of ESC-RPCs for subpopulations expressing PAX6, RAX, OTX2, and NRL, respectively. (D) H&E staining of the section of an ocular tumor 3 weeks after ESC-RPC injection. (E) The magnified image of tumor tissue in D. (F) A cryosection of a transplanted eyeball containing the tumor tissue from injected ESC-RPCsEGFP (green). (G) Protein levels of several markers in tumor tissues were compared with those in the normal adult retina. (H) The amplitude of ERG b waves at different light intensities is shown for normal mice and SI-treated mice after subretinal injection of P-RPCs, ESC-RPCs, and the cell culture medium (Sham injection), respectively. Mean ± SD; ANOVA; **P < 0.01; n = 10 for each group. Scale bar: 25 μm (A); 500 μm (D); 100 μm (E and F).