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Long-term safety issues of iPSC-based cell therapy in a spinal cord injury model: oncogenic transformation with epithelial-mesenchymal transition.

Nori S, Okada Y, Nishimura S, Sasaki T, Itakura G, Kobayashi Y, Renault-Mihara F, Shimizu A, Koya I, Yoshida R, Kudoh J, Koike M, Uchiyama Y, Ikeda E, Toyama Y, Nakamura M, Okano H - Stem Cell Reports (2015)

Bottom Line: However, long-term observation (for up to 103 days) revealed deteriorated motor function accompanied by tumor formation.The tumors consisted of Nestin(+) undifferentiated neural cells and exhibited activation of the OCT4 transgene.Transcriptome analysis revealed that a heightened mesenchymal transition may have contributed to the progression of tumors derived from grafted cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedic Surgery, Keio University School of Medicine, 35 Shinanomachi, Shinjuku, Tokyo 160-8582, Japan; Department of Physiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku, Tokyo 160-8582, Japan.

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Histological and Gene-Expression Analyses of Tumors(A) Schematic of histological analyses of tumors.(B) Correlation between tumor diameter and the proportion of grafted cells that were Nestin+ at 47 and 103 days post-transplantation (TP).(C) Correlation between tumor diameter and the proportion of grafted cells that were Ki-67+.(D) Correlation between tumor diameter and the proportion of grafted cells that were OCT4+.(E) Correlation between the tumor diameter and the proportion of grafted cells that were OCT4+/HNu+ at 103 days after TP.(F) Correlation between the number of days after TP (47 or 103 days after TP) and the proportion of grafted cells that were OCT4+. In (B)–(F), n indicates the number of mice.(G) Schematic of mRNA expression analyses of tumors.(H–M) The expression of human OCT4-Tg, OCT4-Endo, SOX2-Tg, SOX2-Endo, KLF4-Tg, KLF4-Endo, c-MYC-Tg, and c-MYC-Endo mRNA in 253G1 cells, 253G1-NSs, 103-day post-transplant 253G1-NSs (TP 103d), and adult human dermal fibroblasts (HDFs) was analyzed by RT-PCR. Data are presented as expression levels relative to the control (HDFs) 11 days after retroviral transduction of OCT4, SOX2, KLF4, and c-MYC. Values represent the means ± SEM (n = 3 independent experiments).The p values shown in (B)–(F) were calculated using Scheffe’s test, and p values to determine significance were calculated using the Kruskal-Wallis non-parametric test: (B) 5.00E-06, (C) 7.20E-06, (D) 0.01, and (F) 1.33E-04. ∗p < 0.05, ∗∗p < 0.01. ns, non-significant.
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fig3: Histological and Gene-Expression Analyses of Tumors(A) Schematic of histological analyses of tumors.(B) Correlation between tumor diameter and the proportion of grafted cells that were Nestin+ at 47 and 103 days post-transplantation (TP).(C) Correlation between tumor diameter and the proportion of grafted cells that were Ki-67+.(D) Correlation between tumor diameter and the proportion of grafted cells that were OCT4+.(E) Correlation between the tumor diameter and the proportion of grafted cells that were OCT4+/HNu+ at 103 days after TP.(F) Correlation between the number of days after TP (47 or 103 days after TP) and the proportion of grafted cells that were OCT4+. In (B)–(F), n indicates the number of mice.(G) Schematic of mRNA expression analyses of tumors.(H–M) The expression of human OCT4-Tg, OCT4-Endo, SOX2-Tg, SOX2-Endo, KLF4-Tg, KLF4-Endo, c-MYC-Tg, and c-MYC-Endo mRNA in 253G1 cells, 253G1-NSs, 103-day post-transplant 253G1-NSs (TP 103d), and adult human dermal fibroblasts (HDFs) was analyzed by RT-PCR. Data are presented as expression levels relative to the control (HDFs) 11 days after retroviral transduction of OCT4, SOX2, KLF4, and c-MYC. Values represent the means ± SEM (n = 3 independent experiments).The p values shown in (B)–(F) were calculated using Scheffe’s test, and p values to determine significance were calculated using the Kruskal-Wallis non-parametric test: (B) 5.00E-06, (C) 7.20E-06, (D) 0.01, and (F) 1.33E-04. ∗p < 0.05, ∗∗p < 0.01. ns, non-significant.

Mentions: Next, we examined the correlation between tumor diameter and the percentages of Nestin+, Ki-67+, or OCT4+ cells among HNu+ grafted cells (Figure 3A). Statistical analysis revealed a significant correlation between tumor diameter and the percentage of Nestin+/HNu+ cells (Figure 3B). Compared with the small-tumor group, the large- and medium-tumor groups contained significantly higher percentages of Ki-67+/HNu+ cells and OCT4+/HNu+ cells (Figures 3C and 3D). However, there was no significant correlation between tumor diameter and the percentage of OCT4+/HNu+ cells at 103 days post-transplantation (Figure 3E). Meanwhile, significantly more OCT4+/HNu+ cells were observed at 103 days than at 47 days (Figure 3F); thus, the percentage of OCT4+/HNu+ cells correlated positively with post-transplant duration.


Long-term safety issues of iPSC-based cell therapy in a spinal cord injury model: oncogenic transformation with epithelial-mesenchymal transition.

Nori S, Okada Y, Nishimura S, Sasaki T, Itakura G, Kobayashi Y, Renault-Mihara F, Shimizu A, Koya I, Yoshida R, Kudoh J, Koike M, Uchiyama Y, Ikeda E, Toyama Y, Nakamura M, Okano H - Stem Cell Reports (2015)

Histological and Gene-Expression Analyses of Tumors(A) Schematic of histological analyses of tumors.(B) Correlation between tumor diameter and the proportion of grafted cells that were Nestin+ at 47 and 103 days post-transplantation (TP).(C) Correlation between tumor diameter and the proportion of grafted cells that were Ki-67+.(D) Correlation between tumor diameter and the proportion of grafted cells that were OCT4+.(E) Correlation between the tumor diameter and the proportion of grafted cells that were OCT4+/HNu+ at 103 days after TP.(F) Correlation between the number of days after TP (47 or 103 days after TP) and the proportion of grafted cells that were OCT4+. In (B)–(F), n indicates the number of mice.(G) Schematic of mRNA expression analyses of tumors.(H–M) The expression of human OCT4-Tg, OCT4-Endo, SOX2-Tg, SOX2-Endo, KLF4-Tg, KLF4-Endo, c-MYC-Tg, and c-MYC-Endo mRNA in 253G1 cells, 253G1-NSs, 103-day post-transplant 253G1-NSs (TP 103d), and adult human dermal fibroblasts (HDFs) was analyzed by RT-PCR. Data are presented as expression levels relative to the control (HDFs) 11 days after retroviral transduction of OCT4, SOX2, KLF4, and c-MYC. Values represent the means ± SEM (n = 3 independent experiments).The p values shown in (B)–(F) were calculated using Scheffe’s test, and p values to determine significance were calculated using the Kruskal-Wallis non-parametric test: (B) 5.00E-06, (C) 7.20E-06, (D) 0.01, and (F) 1.33E-04. ∗p < 0.05, ∗∗p < 0.01. ns, non-significant.
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fig3: Histological and Gene-Expression Analyses of Tumors(A) Schematic of histological analyses of tumors.(B) Correlation between tumor diameter and the proportion of grafted cells that were Nestin+ at 47 and 103 days post-transplantation (TP).(C) Correlation between tumor diameter and the proportion of grafted cells that were Ki-67+.(D) Correlation between tumor diameter and the proportion of grafted cells that were OCT4+.(E) Correlation between the tumor diameter and the proportion of grafted cells that were OCT4+/HNu+ at 103 days after TP.(F) Correlation between the number of days after TP (47 or 103 days after TP) and the proportion of grafted cells that were OCT4+. In (B)–(F), n indicates the number of mice.(G) Schematic of mRNA expression analyses of tumors.(H–M) The expression of human OCT4-Tg, OCT4-Endo, SOX2-Tg, SOX2-Endo, KLF4-Tg, KLF4-Endo, c-MYC-Tg, and c-MYC-Endo mRNA in 253G1 cells, 253G1-NSs, 103-day post-transplant 253G1-NSs (TP 103d), and adult human dermal fibroblasts (HDFs) was analyzed by RT-PCR. Data are presented as expression levels relative to the control (HDFs) 11 days after retroviral transduction of OCT4, SOX2, KLF4, and c-MYC. Values represent the means ± SEM (n = 3 independent experiments).The p values shown in (B)–(F) were calculated using Scheffe’s test, and p values to determine significance were calculated using the Kruskal-Wallis non-parametric test: (B) 5.00E-06, (C) 7.20E-06, (D) 0.01, and (F) 1.33E-04. ∗p < 0.05, ∗∗p < 0.01. ns, non-significant.
Mentions: Next, we examined the correlation between tumor diameter and the percentages of Nestin+, Ki-67+, or OCT4+ cells among HNu+ grafted cells (Figure 3A). Statistical analysis revealed a significant correlation between tumor diameter and the percentage of Nestin+/HNu+ cells (Figure 3B). Compared with the small-tumor group, the large- and medium-tumor groups contained significantly higher percentages of Ki-67+/HNu+ cells and OCT4+/HNu+ cells (Figures 3C and 3D). However, there was no significant correlation between tumor diameter and the percentage of OCT4+/HNu+ cells at 103 days post-transplantation (Figure 3E). Meanwhile, significantly more OCT4+/HNu+ cells were observed at 103 days than at 47 days (Figure 3F); thus, the percentage of OCT4+/HNu+ cells correlated positively with post-transplant duration.

Bottom Line: However, long-term observation (for up to 103 days) revealed deteriorated motor function accompanied by tumor formation.The tumors consisted of Nestin(+) undifferentiated neural cells and exhibited activation of the OCT4 transgene.Transcriptome analysis revealed that a heightened mesenchymal transition may have contributed to the progression of tumors derived from grafted cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedic Surgery, Keio University School of Medicine, 35 Shinanomachi, Shinjuku, Tokyo 160-8582, Japan; Department of Physiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku, Tokyo 160-8582, Japan.

Show MeSH
Related in: MedlinePlus