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Erythropoietin improves the accumulation and therapeutic effects of carboplatin by enhancing tumor vascularization and perfusion.

Doleschel D, Rix A, Arns S, Palmowski K, Gremse F, Merkle R, Salopiata F, Klingmüller U, Jarsch M, Kiessling F, Lederle W - Theranostics (2015)

Bottom Line: In both xenografts, rhuEpo co-medication significantly increased vessel densities, diameters and the amount of perfused vessels.However, compared with solely carboplatin-treated tumors, tumor growth was significantly slower in the groups co-medicated with rhuEpo.Doses and indications may be personalized and refined using theranostic EpoR-probes.

View Article: PubMed Central - PubMed

Affiliation: 1. Experimental Molecular Imaging, Medical Faculty, RWTH Aachen University, Aachen, Germany.

ABSTRACT
Recombinant human erythropoietin (rhuEpo) is currently under debate for the treatment of chemotherapy-induced anemia due to clinical trials showing adverse effects in Epo-treated patients and the discovery of the erythropoietin-receptor (EpoR) in tumor and endothelial cells. Here, using Epo-Cy5.5 as theranostic near-infrared fluorescent probe we analyzed the effects of rhuEpo as co-medication to carboplatin in non-small-cell-lung-cancer (NSCLC)-xenografts with different tumor cell EpoR-expression (H838 ~8-fold higher than A549). Nude mice bearing subcutaneous A549 and H838 NSCLC-xenografts received either only carboplatin or carboplatin and co-medication of rhuEpo in two different doses. Tumor sizes and relative blood volumes (rBV) were longitudinally measured by 3D-contrast-enhanced ultrasound (3D-US). Tumoral EpoR-levels were determined by combined fluorescence molecular tomography (FMT)/ micro computed tomography (µCT) hybrid imaging. We found that rhuEpo predominantly acted on the tumor endothelium. In both xenografts, rhuEpo co-medication significantly increased vessel densities, diameters and the amount of perfused vessels. Accordingly, rhuEpo induced EpoR-phoshorylation and stimulated proliferation of endothelial cells. However, compared with solely carboplatin-treated tumors, tumor growth was significantly slower in the groups co-medicated with rhuEpo. This is explained by the Epo-mediated vascular remodeling leading to improved drug delivery as obvious by a more than 2-fold higher carboplatin accumulation and significantly enhanced tumor apoptosis. In addition, co-medication of rhuEpo reduced tumor hypoxia and diminished intratumoral EpoR-levels which continuously increased during carboplatin (Cp) -treatment. These findings suggest that co-medication of rhuEpo in well balanced doses can be used to improve the accumulation of anticancer drugs. Doses and indications may be personalized and refined using theranostic EpoR-probes.

No MeSH data available.


Related in: MedlinePlus

Epo co-medication has differential effects on EpoR-expression but reduces tumor hypoxia. The EpoR-status in the tumors was longitudinally assessed by µCT/FMT hybrid imaging using Epo-Cy5.5. A: µCT/FMT fusion image demonstrating higher Epo-Cy5.5 accumulation in only carboplatin-treated (Cp) H838 (left panel) versus tumors additionally treated with 20 µg/kg of Epo (Cp + Epo 20, right panel) after three weeks of treatment. Note the different signal intensities in the tumors (lined in red), whereas they are comparable in the bone marrow of pelvis and tibia (arrowheads). B: 3D rendering of reconstructed µCT/FMT-data of the abdominal part of a H838 tumor-bearing mouse at day 21. Specific accumulation of Epo-Cy5.5 (blue) is visible in the EpoR-positive organs, the bone marrow of pelvis, spinal cord and hind legs (white), the tumor (green) and the kidneys (yellow). C: Epo-Cy5.5 levels show a similar increase in carboplatin- and additionally Epo-treated A549 tumors (upper graph), whereas in H838, significantly lower levels are recorded for combinatorial Epo-treated compared with only carboplatin-treated tumors in the third week of therapy (lower graph) (n = 5, *p < 0.05). D: Immunostainings for HIF-1α (red), endothelial cells (CD31, green) and cell nuclei (DAPI, blue) of representative A549 tumor sections at day 21. Note the reduced hypoxia in the additionally Epo-treated groups (scale bars: 200 µm). E: Quantification of HIF-1α staining reveals a significantly lower hypoxia in additionally Epo-treated A549 and H838 tumors at day 21 (n = 5, *p < 0.05). Cp: carboplatin, Epo 5: rhuEpo 5 µg/kg, Epo 20: rhuEpo 20 µg/kg.
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Figure 6: Epo co-medication has differential effects on EpoR-expression but reduces tumor hypoxia. The EpoR-status in the tumors was longitudinally assessed by µCT/FMT hybrid imaging using Epo-Cy5.5. A: µCT/FMT fusion image demonstrating higher Epo-Cy5.5 accumulation in only carboplatin-treated (Cp) H838 (left panel) versus tumors additionally treated with 20 µg/kg of Epo (Cp + Epo 20, right panel) after three weeks of treatment. Note the different signal intensities in the tumors (lined in red), whereas they are comparable in the bone marrow of pelvis and tibia (arrowheads). B: 3D rendering of reconstructed µCT/FMT-data of the abdominal part of a H838 tumor-bearing mouse at day 21. Specific accumulation of Epo-Cy5.5 (blue) is visible in the EpoR-positive organs, the bone marrow of pelvis, spinal cord and hind legs (white), the tumor (green) and the kidneys (yellow). C: Epo-Cy5.5 levels show a similar increase in carboplatin- and additionally Epo-treated A549 tumors (upper graph), whereas in H838, significantly lower levels are recorded for combinatorial Epo-treated compared with only carboplatin-treated tumors in the third week of therapy (lower graph) (n = 5, *p < 0.05). D: Immunostainings for HIF-1α (red), endothelial cells (CD31, green) and cell nuclei (DAPI, blue) of representative A549 tumor sections at day 21. Note the reduced hypoxia in the additionally Epo-treated groups (scale bars: 200 µm). E: Quantification of HIF-1α staining reveals a significantly lower hypoxia in additionally Epo-treated A549 and H838 tumors at day 21 (n = 5, *p < 0.05). Cp: carboplatin, Epo 5: rhuEpo 5 µg/kg, Epo 20: rhuEpo 20 µg/kg.

Mentions: In order to analyze whether Epo co-medication had an effect on EpoR-expression, the EpoR-levels were measured longitudinally by µCT/FMT hybrid imaging using the NIR-labeled EpoR-probe Epo-Cy5.5 26. In both NSCLC-models, the Epo-Cy5.5 levels continuously increased during sole carboplatin-treatment, pointing to an up-regulation of EpoR in the tumor tissue (Fig. 6 C, Supplementary Fig. 4). However, in the H838 model, Epo-Cy5.5 accumulation was markedly lower in tumors co-medicated with Epo, reaching significant differences to solely carboplatin-treated tumors in week three of therapy (Fig. 6 A - C, *p < 0.05). A marginal decrease in EpoR-expression was also observed in the Epo co-medicated A549 tumors (Fig. 6 C, Supplementary Fig. 4). Since the tumor vascularization was significantly higher in the Epo co-medicated groups, which could have facilitated passive probe accumulation, the reduced Epo-Cy5.5 levels cannot be explained by a pharmacological effect but rather by down-regulation of EpoR.


Erythropoietin improves the accumulation and therapeutic effects of carboplatin by enhancing tumor vascularization and perfusion.

Doleschel D, Rix A, Arns S, Palmowski K, Gremse F, Merkle R, Salopiata F, Klingmüller U, Jarsch M, Kiessling F, Lederle W - Theranostics (2015)

Epo co-medication has differential effects on EpoR-expression but reduces tumor hypoxia. The EpoR-status in the tumors was longitudinally assessed by µCT/FMT hybrid imaging using Epo-Cy5.5. A: µCT/FMT fusion image demonstrating higher Epo-Cy5.5 accumulation in only carboplatin-treated (Cp) H838 (left panel) versus tumors additionally treated with 20 µg/kg of Epo (Cp + Epo 20, right panel) after three weeks of treatment. Note the different signal intensities in the tumors (lined in red), whereas they are comparable in the bone marrow of pelvis and tibia (arrowheads). B: 3D rendering of reconstructed µCT/FMT-data of the abdominal part of a H838 tumor-bearing mouse at day 21. Specific accumulation of Epo-Cy5.5 (blue) is visible in the EpoR-positive organs, the bone marrow of pelvis, spinal cord and hind legs (white), the tumor (green) and the kidneys (yellow). C: Epo-Cy5.5 levels show a similar increase in carboplatin- and additionally Epo-treated A549 tumors (upper graph), whereas in H838, significantly lower levels are recorded for combinatorial Epo-treated compared with only carboplatin-treated tumors in the third week of therapy (lower graph) (n = 5, *p < 0.05). D: Immunostainings for HIF-1α (red), endothelial cells (CD31, green) and cell nuclei (DAPI, blue) of representative A549 tumor sections at day 21. Note the reduced hypoxia in the additionally Epo-treated groups (scale bars: 200 µm). E: Quantification of HIF-1α staining reveals a significantly lower hypoxia in additionally Epo-treated A549 and H838 tumors at day 21 (n = 5, *p < 0.05). Cp: carboplatin, Epo 5: rhuEpo 5 µg/kg, Epo 20: rhuEpo 20 µg/kg.
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Related In: Results  -  Collection

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Figure 6: Epo co-medication has differential effects on EpoR-expression but reduces tumor hypoxia. The EpoR-status in the tumors was longitudinally assessed by µCT/FMT hybrid imaging using Epo-Cy5.5. A: µCT/FMT fusion image demonstrating higher Epo-Cy5.5 accumulation in only carboplatin-treated (Cp) H838 (left panel) versus tumors additionally treated with 20 µg/kg of Epo (Cp + Epo 20, right panel) after three weeks of treatment. Note the different signal intensities in the tumors (lined in red), whereas they are comparable in the bone marrow of pelvis and tibia (arrowheads). B: 3D rendering of reconstructed µCT/FMT-data of the abdominal part of a H838 tumor-bearing mouse at day 21. Specific accumulation of Epo-Cy5.5 (blue) is visible in the EpoR-positive organs, the bone marrow of pelvis, spinal cord and hind legs (white), the tumor (green) and the kidneys (yellow). C: Epo-Cy5.5 levels show a similar increase in carboplatin- and additionally Epo-treated A549 tumors (upper graph), whereas in H838, significantly lower levels are recorded for combinatorial Epo-treated compared with only carboplatin-treated tumors in the third week of therapy (lower graph) (n = 5, *p < 0.05). D: Immunostainings for HIF-1α (red), endothelial cells (CD31, green) and cell nuclei (DAPI, blue) of representative A549 tumor sections at day 21. Note the reduced hypoxia in the additionally Epo-treated groups (scale bars: 200 µm). E: Quantification of HIF-1α staining reveals a significantly lower hypoxia in additionally Epo-treated A549 and H838 tumors at day 21 (n = 5, *p < 0.05). Cp: carboplatin, Epo 5: rhuEpo 5 µg/kg, Epo 20: rhuEpo 20 µg/kg.
Mentions: In order to analyze whether Epo co-medication had an effect on EpoR-expression, the EpoR-levels were measured longitudinally by µCT/FMT hybrid imaging using the NIR-labeled EpoR-probe Epo-Cy5.5 26. In both NSCLC-models, the Epo-Cy5.5 levels continuously increased during sole carboplatin-treatment, pointing to an up-regulation of EpoR in the tumor tissue (Fig. 6 C, Supplementary Fig. 4). However, in the H838 model, Epo-Cy5.5 accumulation was markedly lower in tumors co-medicated with Epo, reaching significant differences to solely carboplatin-treated tumors in week three of therapy (Fig. 6 A - C, *p < 0.05). A marginal decrease in EpoR-expression was also observed in the Epo co-medicated A549 tumors (Fig. 6 C, Supplementary Fig. 4). Since the tumor vascularization was significantly higher in the Epo co-medicated groups, which could have facilitated passive probe accumulation, the reduced Epo-Cy5.5 levels cannot be explained by a pharmacological effect but rather by down-regulation of EpoR.

Bottom Line: In both xenografts, rhuEpo co-medication significantly increased vessel densities, diameters and the amount of perfused vessels.However, compared with solely carboplatin-treated tumors, tumor growth was significantly slower in the groups co-medicated with rhuEpo.Doses and indications may be personalized and refined using theranostic EpoR-probes.

View Article: PubMed Central - PubMed

Affiliation: 1. Experimental Molecular Imaging, Medical Faculty, RWTH Aachen University, Aachen, Germany.

ABSTRACT
Recombinant human erythropoietin (rhuEpo) is currently under debate for the treatment of chemotherapy-induced anemia due to clinical trials showing adverse effects in Epo-treated patients and the discovery of the erythropoietin-receptor (EpoR) in tumor and endothelial cells. Here, using Epo-Cy5.5 as theranostic near-infrared fluorescent probe we analyzed the effects of rhuEpo as co-medication to carboplatin in non-small-cell-lung-cancer (NSCLC)-xenografts with different tumor cell EpoR-expression (H838 ~8-fold higher than A549). Nude mice bearing subcutaneous A549 and H838 NSCLC-xenografts received either only carboplatin or carboplatin and co-medication of rhuEpo in two different doses. Tumor sizes and relative blood volumes (rBV) were longitudinally measured by 3D-contrast-enhanced ultrasound (3D-US). Tumoral EpoR-levels were determined by combined fluorescence molecular tomography (FMT)/ micro computed tomography (µCT) hybrid imaging. We found that rhuEpo predominantly acted on the tumor endothelium. In both xenografts, rhuEpo co-medication significantly increased vessel densities, diameters and the amount of perfused vessels. Accordingly, rhuEpo induced EpoR-phoshorylation and stimulated proliferation of endothelial cells. However, compared with solely carboplatin-treated tumors, tumor growth was significantly slower in the groups co-medicated with rhuEpo. This is explained by the Epo-mediated vascular remodeling leading to improved drug delivery as obvious by a more than 2-fold higher carboplatin accumulation and significantly enhanced tumor apoptosis. In addition, co-medication of rhuEpo reduced tumor hypoxia and diminished intratumoral EpoR-levels which continuously increased during carboplatin (Cp) -treatment. These findings suggest that co-medication of rhuEpo in well balanced doses can be used to improve the accumulation of anticancer drugs. Doses and indications may be personalized and refined using theranostic EpoR-probes.

No MeSH data available.


Related in: MedlinePlus