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Tumor-Free Transplantation of Patient-Derived Induced Pluripotent Stem Cell Progeny for Customized Islet Regeneration

View Article: PubMed Central - PubMed

ABSTRACT

Transplantation of progenitors from induced pluripotent stem cells reprogrammed by lentiviral vectors led to the formation of invasive teratocarcinoma-like tumors in more than 90% of immunodeficient mice. Combined transgene-free reprogramming and elimination of residual pluripotent cells by enzymatic dissociation ensured tumor-free transplantation, ultimately enabling regeneration of type 1 diabetes-specific human islet structures in vivo.

No MeSH data available.


Related in: MedlinePlus

Integrating cMYC vector imposes tumorigenic load in iPSCs. (A): Four groups of SCID-beige mice (n = 3) received pancreatic progenitor cells from control, Lenti-GFP-, Lenti-cMYC- and Lenti-OSKM-superinfected TGF-iPSCs through transplantation into renal capsules of recipient SCID-beige mice. Three weeks after transplantation, left kidneys with iPSC grafts were recovered. Representative images of left kidneys with iPSC grafts are shown. Scale bars indicate 1 cm. (B): The averages of total kidney and graft weights were determined and compared among control, Lenti-GFP-, Lenti-cMYC- and Lenti-OSKM groups. *, p < .05 versus control by t test. Data represent the mean ± SEM. (C): Cross-sections of the junctions between the kidney and iPSC-derived grafts were visualized upon H&E staining. The junctions are indicated by a green line. Scale bars = 200 µm. Abbreviations: GFP, green fluorescent protein; iPSCs, induced pluripotent stem cells; OSKM, OCT4, SOX2, KLF4, and cMYC; TGF-iPSCs, transgene-free induced pluripotent stem cells.
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Figure 4: Integrating cMYC vector imposes tumorigenic load in iPSCs. (A): Four groups of SCID-beige mice (n = 3) received pancreatic progenitor cells from control, Lenti-GFP-, Lenti-cMYC- and Lenti-OSKM-superinfected TGF-iPSCs through transplantation into renal capsules of recipient SCID-beige mice. Three weeks after transplantation, left kidneys with iPSC grafts were recovered. Representative images of left kidneys with iPSC grafts are shown. Scale bars indicate 1 cm. (B): The averages of total kidney and graft weights were determined and compared among control, Lenti-GFP-, Lenti-cMYC- and Lenti-OSKM groups. *, p < .05 versus control by t test. Data represent the mean ± SEM. (C): Cross-sections of the junctions between the kidney and iPSC-derived grafts were visualized upon H&E staining. The junctions are indicated by a green line. Scale bars = 200 µm. Abbreviations: GFP, green fluorescent protein; iPSCs, induced pluripotent stem cells; OSKM, OCT4, SOX2, KLF4, and cMYC; TGF-iPSCs, transgene-free induced pluripotent stem cells.

Mentions: To verify the role of integrated reprogramming factors in the tumorigenicity of LV-iPSC progeny, we modified nontumorigenic T1D-speicific TGF-iPSCs by a series of lentiviral vectors and assessed the influence of lentiviral content on the incidence of tumor formation. We tested four TGF-iPSC-derived lines, including (a) unmodified control, (b) lentivector integration control at multiplicity of infection (MOI) titers of 4 (+ Lenti-GFP), (c) cMYC lentivector integration (+ Lenti-cMYC, at MOI = 4), and (d) all four transcription factor lentivector integration (+ Lenti-OCT4, SOX2, KLF4, and c-MYC [OSKM]; Lenti-OCT4, Lenti-SOX2, Lenti-KLF4, and Lenti-cMYC, at MOI = 4 each). Upon transplantation of iPSC-derived pancreatic progenitors from Lenti-cMYC and Lenti-OSKM iPSCs, recipient mice developed large solid tumors (Fig. 4A, 4B). No notable tumor formation was observed in mice transplanted with control or Lenti-GFP-modified iPSC progeny. Of note, histological analysis of the grafts demonstrated that lentiviral introduction of cMYC led to invasion of the graft/tumor into the kidney cortex structure, although unmodified or Lenti-GFP-modified iPSC grafts showed no sign of invasion (Fig. 4C).


Tumor-Free Transplantation of Patient-Derived Induced Pluripotent Stem Cell Progeny for Customized Islet Regeneration
Integrating cMYC vector imposes tumorigenic load in iPSCs. (A): Four groups of SCID-beige mice (n = 3) received pancreatic progenitor cells from control, Lenti-GFP-, Lenti-cMYC- and Lenti-OSKM-superinfected TGF-iPSCs through transplantation into renal capsules of recipient SCID-beige mice. Three weeks after transplantation, left kidneys with iPSC grafts were recovered. Representative images of left kidneys with iPSC grafts are shown. Scale bars indicate 1 cm. (B): The averages of total kidney and graft weights were determined and compared among control, Lenti-GFP-, Lenti-cMYC- and Lenti-OSKM groups. *, p < .05 versus control by t test. Data represent the mean ± SEM. (C): Cross-sections of the junctions between the kidney and iPSC-derived grafts were visualized upon H&E staining. The junctions are indicated by a green line. Scale bars = 200 µm. Abbreviations: GFP, green fluorescent protein; iPSCs, induced pluripotent stem cells; OSKM, OCT4, SOX2, KLF4, and cMYC; TGF-iPSCs, transgene-free induced pluripotent stem cells.
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Related In: Results  -  Collection

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Figure 4: Integrating cMYC vector imposes tumorigenic load in iPSCs. (A): Four groups of SCID-beige mice (n = 3) received pancreatic progenitor cells from control, Lenti-GFP-, Lenti-cMYC- and Lenti-OSKM-superinfected TGF-iPSCs through transplantation into renal capsules of recipient SCID-beige mice. Three weeks after transplantation, left kidneys with iPSC grafts were recovered. Representative images of left kidneys with iPSC grafts are shown. Scale bars indicate 1 cm. (B): The averages of total kidney and graft weights were determined and compared among control, Lenti-GFP-, Lenti-cMYC- and Lenti-OSKM groups. *, p < .05 versus control by t test. Data represent the mean ± SEM. (C): Cross-sections of the junctions between the kidney and iPSC-derived grafts were visualized upon H&E staining. The junctions are indicated by a green line. Scale bars = 200 µm. Abbreviations: GFP, green fluorescent protein; iPSCs, induced pluripotent stem cells; OSKM, OCT4, SOX2, KLF4, and cMYC; TGF-iPSCs, transgene-free induced pluripotent stem cells.
Mentions: To verify the role of integrated reprogramming factors in the tumorigenicity of LV-iPSC progeny, we modified nontumorigenic T1D-speicific TGF-iPSCs by a series of lentiviral vectors and assessed the influence of lentiviral content on the incidence of tumor formation. We tested four TGF-iPSC-derived lines, including (a) unmodified control, (b) lentivector integration control at multiplicity of infection (MOI) titers of 4 (+ Lenti-GFP), (c) cMYC lentivector integration (+ Lenti-cMYC, at MOI = 4), and (d) all four transcription factor lentivector integration (+ Lenti-OCT4, SOX2, KLF4, and c-MYC [OSKM]; Lenti-OCT4, Lenti-SOX2, Lenti-KLF4, and Lenti-cMYC, at MOI = 4 each). Upon transplantation of iPSC-derived pancreatic progenitors from Lenti-cMYC and Lenti-OSKM iPSCs, recipient mice developed large solid tumors (Fig. 4A, 4B). No notable tumor formation was observed in mice transplanted with control or Lenti-GFP-modified iPSC progeny. Of note, histological analysis of the grafts demonstrated that lentiviral introduction of cMYC led to invasion of the graft/tumor into the kidney cortex structure, although unmodified or Lenti-GFP-modified iPSC grafts showed no sign of invasion (Fig. 4C).

View Article: PubMed Central - PubMed

ABSTRACT

Transplantation of progenitors from induced pluripotent stem cells reprogrammed by lentiviral vectors led to the formation of invasive teratocarcinoma-like tumors in more than 90% of immunodeficient mice. Combined transgene-free reprogramming and elimination of residual pluripotent cells by enzymatic dissociation ensured tumor-free transplantation, ultimately enabling regeneration of type 1 diabetes-specific human islet structures in vivo.

No MeSH data available.


Related in: MedlinePlus