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Growth inhibition of colorectal carcinoma by lentiviral TRAIL-transgenic human mesenchymal stem cells requires their substantial intratumoral presence.

Luetzkendorf J, Mueller LP, Mueller T, Caysa H, Nerger K, Schmoll HJ - J. Cell. Mol. Med. (2010)

Bottom Line: Mesenchymal stem cells (MSC) home to tumours and may therefore serve as a novel therapeutic tool for intratumoral delivery of antineoplastic factors.We generated TRAIL-MSC by transduction of human MSC with a third generation lentiviral vector system and analysed their characteristics and capacity to inhibit CRC growth. (1) TRAIL-MSC showed stable transgene expression with neither changes in the defining MSC characteristics nor signs of malignant transformation. (2) Upon direct in vitro coculture TRAIL-MSC induced apoptosis in TRAIL-sensitive CRC-cell lines (DLD-1 and HCT-15) but also in CRC-cell lines resistant to soluble TRAIL (HCT-8 and SW480). (3) In mixed subcutaneous (s.c.) xenografts TRAIL-MSC inhibited CRC-tumour growth presumably by apoptosis induction but a substantial proportion of TRAIL-MSC within the total tumour cell number was needed to yield such anti-tumour effect. (4) Systemic application of TRAIL-MSC had no effect on the growth of s.c.Systemic TRAIL-MSC caused no toxicity in this model. (5) Wild-type MSC seemed to exert a tumour growth-supporting effect in mixed s.c.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine IV, Oncology/Hematology, Martin-Luther-University Halle-Wittenberg, Halle, Germany.

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Lack of effects of systemic TRAIL-MSC on growth of s.c. CRC xenografts due to poor tumour integration in agreement with the need for a substantial proportion of tumour-integrated TRAIL-MSC to inhibit CRC-tumour growth in vivo. (A) DiI-labelled WT- and TRAIL-MSC were injected into tail vein of nude mice bearing s.c. GFP-DLD-1 xenografts on days 2, 4, 7, 10 and 15. Tumour sizes were quantified by imaging the GFP-fluorescence intensity of xenografts without MSC application (black square), with i.v. WT-MSC (green square) and with i.v. TRAIL-MSC (blue triangle). The total GFP-signal intensity divided by exposure time in ms is plotted. The total intensity / ms 2.5 hrs after generating xenografts was set 100%. Depicted as mean ± standard error of the mean (N= 3), except for tumours without MSC application (N= 1). (B) Lungs from mice with and without i.v. application of DiI-labelled WT- or TRAIL-MSC were dissected and imaged ex vivo to visualize DiI-labelled cells (red). The upper panel shows greyscale images. In the lower panel the respective fluorescence images are pictured. (C) Cryosections from lung and s.c. DLD-1 xenograft after i.v. application of DiI-labelled TRAIL-MSC. Nuclei were counterstained with DAPI. Fluorescence images are pictured as overlays of DiI (red) and DAPI (blue) fluorescence. Original magnification ×400. (D) S.c. mixed xenografts were generated with DsRed-DLD-1 cells and different amounts of WT-MSC (i.e. native or GFP-MSC) or TRAIL-MSC. Tumour size was examined by imaging the DsRed fluorescence intensity. Fluorescence signals of xenografts from DsRed-DLD-1 cells without MSC (black square), from DsRed-DLD-1 cells mixed with WT-MSC (green square) and TRAIL-MSC (blue triangle) were quantified. The total DsRed-signal intensity of tumours divided by exposure time in ms is plotted. The total intensity / ms 2.5 hrs after generating xenografts was set as 100%. Depicted as mean ± standard error of the mean (20% MSC N= 9; 10%N= 6; 3%N= 6; 1%N= 3; no MSC N= 4). Statistical relevance is only reached in comparison of 20%TRAIL-MSC with 20% WT-MSC (P= 0.005). Note that for 20% MSC the curve for WT-MSC is composed of data from xenografts with 20%GFP-MSC (N= 6) and with 20% WT-MSC (N= 3).
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fig06: Lack of effects of systemic TRAIL-MSC on growth of s.c. CRC xenografts due to poor tumour integration in agreement with the need for a substantial proportion of tumour-integrated TRAIL-MSC to inhibit CRC-tumour growth in vivo. (A) DiI-labelled WT- and TRAIL-MSC were injected into tail vein of nude mice bearing s.c. GFP-DLD-1 xenografts on days 2, 4, 7, 10 and 15. Tumour sizes were quantified by imaging the GFP-fluorescence intensity of xenografts without MSC application (black square), with i.v. WT-MSC (green square) and with i.v. TRAIL-MSC (blue triangle). The total GFP-signal intensity divided by exposure time in ms is plotted. The total intensity / ms 2.5 hrs after generating xenografts was set 100%. Depicted as mean ± standard error of the mean (N= 3), except for tumours without MSC application (N= 1). (B) Lungs from mice with and without i.v. application of DiI-labelled WT- or TRAIL-MSC were dissected and imaged ex vivo to visualize DiI-labelled cells (red). The upper panel shows greyscale images. In the lower panel the respective fluorescence images are pictured. (C) Cryosections from lung and s.c. DLD-1 xenograft after i.v. application of DiI-labelled TRAIL-MSC. Nuclei were counterstained with DAPI. Fluorescence images are pictured as overlays of DiI (red) and DAPI (blue) fluorescence. Original magnification ×400. (D) S.c. mixed xenografts were generated with DsRed-DLD-1 cells and different amounts of WT-MSC (i.e. native or GFP-MSC) or TRAIL-MSC. Tumour size was examined by imaging the DsRed fluorescence intensity. Fluorescence signals of xenografts from DsRed-DLD-1 cells without MSC (black square), from DsRed-DLD-1 cells mixed with WT-MSC (green square) and TRAIL-MSC (blue triangle) were quantified. The total DsRed-signal intensity of tumours divided by exposure time in ms is plotted. The total intensity / ms 2.5 hrs after generating xenografts was set as 100%. Depicted as mean ± standard error of the mean (20% MSC N= 9; 10%N= 6; 3%N= 6; 1%N= 3; no MSC N= 4). Statistical relevance is only reached in comparison of 20%TRAIL-MSC with 20% WT-MSC (P= 0.005). Note that for 20% MSC the curve for WT-MSC is composed of data from xenografts with 20%GFP-MSC (N= 6) and with 20% WT-MSC (N= 3).

Mentions: In comparison to controls receiving no MSC a similar tumour growth occurred in mice receiving WT-MSC as well as in mice receiving TRAIL-MSC (Fig. 6A). No signs of toxicity were seen upon systemic transplantation of TRAIL-MSC. In particular no organ dysfunction and no formation of additional tumours were observed upon live observation of animals and macroscopic inspection after killing.


Growth inhibition of colorectal carcinoma by lentiviral TRAIL-transgenic human mesenchymal stem cells requires their substantial intratumoral presence.

Luetzkendorf J, Mueller LP, Mueller T, Caysa H, Nerger K, Schmoll HJ - J. Cell. Mol. Med. (2010)

Lack of effects of systemic TRAIL-MSC on growth of s.c. CRC xenografts due to poor tumour integration in agreement with the need for a substantial proportion of tumour-integrated TRAIL-MSC to inhibit CRC-tumour growth in vivo. (A) DiI-labelled WT- and TRAIL-MSC were injected into tail vein of nude mice bearing s.c. GFP-DLD-1 xenografts on days 2, 4, 7, 10 and 15. Tumour sizes were quantified by imaging the GFP-fluorescence intensity of xenografts without MSC application (black square), with i.v. WT-MSC (green square) and with i.v. TRAIL-MSC (blue triangle). The total GFP-signal intensity divided by exposure time in ms is plotted. The total intensity / ms 2.5 hrs after generating xenografts was set 100%. Depicted as mean ± standard error of the mean (N= 3), except for tumours without MSC application (N= 1). (B) Lungs from mice with and without i.v. application of DiI-labelled WT- or TRAIL-MSC were dissected and imaged ex vivo to visualize DiI-labelled cells (red). The upper panel shows greyscale images. In the lower panel the respective fluorescence images are pictured. (C) Cryosections from lung and s.c. DLD-1 xenograft after i.v. application of DiI-labelled TRAIL-MSC. Nuclei were counterstained with DAPI. Fluorescence images are pictured as overlays of DiI (red) and DAPI (blue) fluorescence. Original magnification ×400. (D) S.c. mixed xenografts were generated with DsRed-DLD-1 cells and different amounts of WT-MSC (i.e. native or GFP-MSC) or TRAIL-MSC. Tumour size was examined by imaging the DsRed fluorescence intensity. Fluorescence signals of xenografts from DsRed-DLD-1 cells without MSC (black square), from DsRed-DLD-1 cells mixed with WT-MSC (green square) and TRAIL-MSC (blue triangle) were quantified. The total DsRed-signal intensity of tumours divided by exposure time in ms is plotted. The total intensity / ms 2.5 hrs after generating xenografts was set as 100%. Depicted as mean ± standard error of the mean (20% MSC N= 9; 10%N= 6; 3%N= 6; 1%N= 3; no MSC N= 4). Statistical relevance is only reached in comparison of 20%TRAIL-MSC with 20% WT-MSC (P= 0.005). Note that for 20% MSC the curve for WT-MSC is composed of data from xenografts with 20%GFP-MSC (N= 6) and with 20% WT-MSC (N= 3).
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fig06: Lack of effects of systemic TRAIL-MSC on growth of s.c. CRC xenografts due to poor tumour integration in agreement with the need for a substantial proportion of tumour-integrated TRAIL-MSC to inhibit CRC-tumour growth in vivo. (A) DiI-labelled WT- and TRAIL-MSC were injected into tail vein of nude mice bearing s.c. GFP-DLD-1 xenografts on days 2, 4, 7, 10 and 15. Tumour sizes were quantified by imaging the GFP-fluorescence intensity of xenografts without MSC application (black square), with i.v. WT-MSC (green square) and with i.v. TRAIL-MSC (blue triangle). The total GFP-signal intensity divided by exposure time in ms is plotted. The total intensity / ms 2.5 hrs after generating xenografts was set 100%. Depicted as mean ± standard error of the mean (N= 3), except for tumours without MSC application (N= 1). (B) Lungs from mice with and without i.v. application of DiI-labelled WT- or TRAIL-MSC were dissected and imaged ex vivo to visualize DiI-labelled cells (red). The upper panel shows greyscale images. In the lower panel the respective fluorescence images are pictured. (C) Cryosections from lung and s.c. DLD-1 xenograft after i.v. application of DiI-labelled TRAIL-MSC. Nuclei were counterstained with DAPI. Fluorescence images are pictured as overlays of DiI (red) and DAPI (blue) fluorescence. Original magnification ×400. (D) S.c. mixed xenografts were generated with DsRed-DLD-1 cells and different amounts of WT-MSC (i.e. native or GFP-MSC) or TRAIL-MSC. Tumour size was examined by imaging the DsRed fluorescence intensity. Fluorescence signals of xenografts from DsRed-DLD-1 cells without MSC (black square), from DsRed-DLD-1 cells mixed with WT-MSC (green square) and TRAIL-MSC (blue triangle) were quantified. The total DsRed-signal intensity of tumours divided by exposure time in ms is plotted. The total intensity / ms 2.5 hrs after generating xenografts was set as 100%. Depicted as mean ± standard error of the mean (20% MSC N= 9; 10%N= 6; 3%N= 6; 1%N= 3; no MSC N= 4). Statistical relevance is only reached in comparison of 20%TRAIL-MSC with 20% WT-MSC (P= 0.005). Note that for 20% MSC the curve for WT-MSC is composed of data from xenografts with 20%GFP-MSC (N= 6) and with 20% WT-MSC (N= 3).
Mentions: In comparison to controls receiving no MSC a similar tumour growth occurred in mice receiving WT-MSC as well as in mice receiving TRAIL-MSC (Fig. 6A). No signs of toxicity were seen upon systemic transplantation of TRAIL-MSC. In particular no organ dysfunction and no formation of additional tumours were observed upon live observation of animals and macroscopic inspection after killing.

Bottom Line: Mesenchymal stem cells (MSC) home to tumours and may therefore serve as a novel therapeutic tool for intratumoral delivery of antineoplastic factors.We generated TRAIL-MSC by transduction of human MSC with a third generation lentiviral vector system and analysed their characteristics and capacity to inhibit CRC growth. (1) TRAIL-MSC showed stable transgene expression with neither changes in the defining MSC characteristics nor signs of malignant transformation. (2) Upon direct in vitro coculture TRAIL-MSC induced apoptosis in TRAIL-sensitive CRC-cell lines (DLD-1 and HCT-15) but also in CRC-cell lines resistant to soluble TRAIL (HCT-8 and SW480). (3) In mixed subcutaneous (s.c.) xenografts TRAIL-MSC inhibited CRC-tumour growth presumably by apoptosis induction but a substantial proportion of TRAIL-MSC within the total tumour cell number was needed to yield such anti-tumour effect. (4) Systemic application of TRAIL-MSC had no effect on the growth of s.c.Systemic TRAIL-MSC caused no toxicity in this model. (5) Wild-type MSC seemed to exert a tumour growth-supporting effect in mixed s.c.

View Article: PubMed Central - PubMed

Affiliation: Department of Internal Medicine IV, Oncology/Hematology, Martin-Luther-University Halle-Wittenberg, Halle, Germany.

Show MeSH
Related in: MedlinePlus