The online version of this article (https://doi.org/10.1186/s13046-018-0671-0) contains supplementary material, which is available to authorized users.
Small molecule ONC201 is an investigational anti-tumor agent that upregulates intra-tumoral TRAIL expression and the integrated stress response pathway. A Phase I clinical trial using ONC201 therapy in advanced cancer patients has been completed and the drug has progressed into Phase II trials in several cancer types. Colorectal cancer (CRC) remains one of the leading causes of cancer worldwide and metastatic disease has a poor prognosis. Clinical trials in CRC and other tumor types have demonstrated that therapeutics targeting the vascular endothelial growth factor (VEGF) pathway, such as bevacizumab, are effective in combination with certain chemotherapeutic agents.
We investigated the potential combination of VEGF inhibitors such as bevacizumab and its murine-counterpart; along with other anti-angiogenic agents and ONC201 in both CRC xenograft and patient-derived xenograft (PDX) models. We utilized non-invasive imaging and immunohistochemistry to determine potential mechanisms of action.
Our results demonstrate significant tumor regression or complete tumor ablation in human xenografts with the combination of ONC201 with bevacizumab, and in syngeneic MC38 colorectal cancer xenografts using a murine VEGF-A inhibitor. Imaging demonstrated the impact of this combination on decreasing tumor growth and tumor metastasis. Our results indicate that ONC201 and anti-angiogenic agents act through distinct mechanisms while increasing tumor cell death and inhibiting proliferation.
With the use of both a murine VEGF inhibitor in syngeneic models, and bevacizumab in human cell line-derived xenografts, we demonstrate that ONC201 in combination with anti-angiogenic therapies such as bevacizumab represents a promising approach for further testing in the clinic for the treatment of CRC.
Additional file 1: Figure S1. ONC201 and combination with CRC approved chemotherapeutics; CTG data. CTG data of HCT116 CRC cells treated with indicated doses and compounds for 72 hours. (PPTX 80 kb)13046_2018_671_MOESM1_ESM.pptx
Additional file 2: Figure S2. Total number of metastases within liver and lung. A) Total number of metastases in HT29 tissues as seen in H&E slides by pathologist of lungs and liver. B) Total number of metastases seen in HCT116-GFP mice as seen by bioluminescent imaging and pathology of lungs and liver. (PPTX 75 kb)13046_2018_671_MOESM2_ESM.pptx
Additional file 3: Figure S3. ONC201 and combination with bevacizumab are non-toxic in vivo. Blood chemistry panel results from mice treated with indicated drugs. Blood was collected by cardiac puncture at end of experiment. ONC201 50mg/kg weekly. Bevacizumab is 5 mg/kg every 2 weeks. Regorafenib: 5 mg/kg daily. N=3. (PPTX 98 kb)13046_2018_671_MOESM3_ESM.pptx
Additional file 4: Figure S4. VEGF expression in HCT116 xenografts. Representative IHC staining of VEGF expression from mice treated with indicated drugs. Tumors harvested and placed in paraffin. ONC201: 50 mg/kg every week. Regorafenib: 5 mg/kg daily. N=5 tumors, minimum of 3 sections per tumor stained. (PPTX 195 kb)13046_2018_671_MOESM4_ESM.pptx
Additional file 5: Figure S5. Analysis of regorafenib and ONC201 mechanism in combination. A) HUVEC representative images of sprouting from HUVECs grown on Matrigel. B) Quantitation of HUVEC sprouting and branching after 12 hours. C) HCT116 cells from live cell imaging using CHOP-800 and Actin-700 on LiCor Odyssey. Cells treated for 48 hours. ONC201: 5 μM. HUVECS N=4, ONC201 treatment 5 μM, Regorafenib 1 mg/ml. (PPTX 623 kb)13046_2018_671_MOESM5_ESM.pptx
Additional file 6: Figure S6. Full imaging of Superhance blood flow and GFP from HCT116-GFP mice. Representative image using superhance 680 probe in HCT116 xenograft bearing athymic nude mice after 4 weeks. ONC201: 50 mg/kg every week. Regorafenib: 5 mg/kg daily. Bevacizumab: 5 mg/kg every other week. N=5. (PPTX 469 kb)13046_2018_671_MOESM6_ESM.pptx
Additional file 7: Figure S7. CD31 expression in HT29 xenografts. Representative IHC staining of CD31 expression from mice treated with indicated drugs. Tumors harvested and placed in paraffin. ONC201: 50 mg/kg every week. Bevacizumab: 5 mg/kg every other week. N=5 tumors, minimum of 3 sections per tumor stained. (PPTX 184 kb)13046_2018_671_MOESM7_ESM.pptx
Additional file 8: Figure S8. CD31 expression in MC38 CRC tumors. Representative IHC staining of CD31 expression from mice treated with indicated drugs. Tumors harvested and placed in paraffin. ONC201: 50 mg/kg every week. Anti VEGF-A (VEGF inhibitor): 10 μg twice weekly. N=6 tumors, minimum of 3 sections per tumor stained. (PPTX 199 kb)13046_2018_671_MOESM8_ESM.pptx
Additional file 9: Figure S9. TRAIL and M30 ELISA standard curves. Standard curve of both TRAIL and M30 Elisa kits using the manufacturer’s instructions. (PPTX 66 kb)13046_2018_671_MOESM9_ESM.pptx
Additional file 10: Figure S10. Ki67 staining of HCT116 xenografts. Representative IHC staining of Ki67 expression from mice treated with indicated drugs. Tumors harvested and placed in paraffin. ONC201: 50 mg/kg every week. Bevacizumab: 5 mg/kg every other week. Regorafenib: 5 mg/kg daily N=5 tumors, minimum of 3 sections per tumor stained. (PPTX 186 kb)13046_2018_671_MOESM10_ESM.pptx
Additional file 11: Figure S11. Ki67 staining of HT29 xenografts. A) Representative IHC staining B) Quantitation using vectra and Inform analysis. Tumors harvested and placed in paraffin. ONC201: 50 mg/kg every week. Bevacizumab: 5 mg/kg every other week. N=5 tumors, minimum of 3 sections per tumor stained. (PPTX 1470 kb)13046_2018_671_MOESM11_ESM.pptx
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- Anti-tumor effects of ONC201 in combination with VEGF-inhibitors significantly impacts colorectal cancer growth and survival in vivo through complementary non-overlapping mechanisms
C. Leah Kline
Wafik S. El-Deiry
- BioMed Central
Journal of Experimental & Clinical Cancer Research
Elektronische ISSN: 1756-9966
Neu im Fachgebiet Onkologie
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