Search

Supplementary Figure Legends PDF file - 92K, This file contains the Figure legends to Supplementary Figures 1-5

Supplementary Figure 2 PDF file - 394K,Data are presented to show that congenital expression of NRASG12D in mouse melanocytes does not affect eyes or heart development

Supplementary Figure 4 PDF file - 39K, Sequencing of RT-PCR amplified cDNA is presented to show that NRASG12D is expressed in the CNS tumors of our mice

Supplementary Figure 5 PDF file - 1328K, Histopathology is shown to confirm that the tumors from patients 1 and 2 are melanoma of the CNS and that the congenital nevus from patient 2 expresses NRASQ61K

Supplementary Figure 1 Growth kinetics for all 786-0 xenografts included in the analysis from Figure 1; Supplementary Figure 2 Details of the in vitro angiogenesis assay; Supplementary Figure 3 Isolation of a sunitinib-refractory sub-line (786-0-R) from the parental 786-0 cell line; Supplementary Figure 4 Anti-angiogenic activity of sunitinib combined with trametinib within an in vitro tubule formation assay; Supplementary Figure 5 Quantification of tumor vessel density in treated tumors; Supplementary Table 1 Genotyping of the original patient sample and each generation of the PDX by STR typing

Supplementary Figure 1 PDF file - 795K, Data are presented to show that NRASG12D does not induce melanoma when expressed in the melanocytes of adult mice

Sunitinib and pazopanib are antiangiogenic tyrosine kinase inhibitors (TKI) used to treat metastatic renal cell carcinoma (RCC). However, the ability of these drugs to extend progression-free and overall survival in this patient population is limited by drug resistance. It is possible that treatment outcomes in RCC patients could be improved by rationally combining TKIs with other agents. Here, we address whether inhibition of the Ras-Raf-MEK-ERK1/2 pathway is a rational means to improve the response to TKIs in RCC. Using a xenograft model of RCC, we found that tumors that are resistant to sunitinib have a significantly increased angiogenic response compared with tumors that are sensitive to sunitinib in vivo. We also observed significantly increased levels of phosphorylated ERK1/2 in the vasculature of resistant tumors, when compared with sensitive tumors. These data suggested that the Ras-Raf-MEK-ERK1/2 pathway, an important driver of angiogenesis in endothelial cells, remains active in the vasculature of TKI-resistant tumors. Using an in vitro angiogenesis assay, we identified that the MEK inhibitor (MEKI) trametinib has potent antiangiogenic activity. We then show that, when trametinib is combined with a TKI in vivo, more effective suppression of tumor growth and tumor angiogenesis is achieved than when either drug is utilized alone. In conclusion, we provide preclinical evidence that combining a TKI, such as sunitinib or pazopanib, with a MEKI, such as trametinib, is a rational and efficacious treatment regimen for RCC. Mol Cancer Ther; 15(1); 172–83. ©2015 AACR.

Supplementary Figure 3 PDF file - 498K, Histopathology data are presented to confirm that CNS tumors in our mice are malignant melanomas

Increased stiffness in the extracellular matrix (ECM) contributes to tumor progression and metastasis. Therefore, stromal modulating therapies and accompanying biomarkers are being developed to target ECM stiffness. Magnetic resonance (MR) elastography can noninvasively and quantitatively map the viscoelastic properties of tumors in vivo and thus has clear clinical applications. Herein, we used MR elastography, coupled with computational histopathology, to interrogate the contribution of collagen to the tumor biomechanical phenotype and to evaluate its sensitivity to collagenase-induced stromal modulation. Elasticity (Gd) and viscosity (Gl) were significantly greater for orthotopic BT-474 (Gd = 5.9 ± 0.2 kPa, Gl = 4.7 ± 0.2 kPa, n = 7) and luc-MDA-MB-231-LM2-4 (Gd = 7.9 ± 0.4 kPa, Gl = 6.0 ± 0.2 kPa, n = 6) breast cancer xenografts, and luc-PANC1 (Gd = 6.9 ± 0.3 kPa, Gl = 6.2 ± 0.2 kPa, n = 7) pancreatic cancer xenografts, compared with tumors associated with the nervous system, including GTML/Trp53KI/KI medulloblastoma (Gd = 3.5 ± 0.2 kPa, Gl = 2.3 ± 0.2 kPa, n = 7), orthotopic luc-D-212-MG (Gd = 3.5 ± 0.2 kPa, Gl = 2.3 ± 0.2 kPa, n = 7), luc-RG2 (Gd = 3.5 ± 0.2 kPa, Gl = 2.3 ± 0.2 kPa, n = 5), and luc-U-87-MG (Gd = 3.5 ± 0.2 kPa, Gl = 2.3 ± 0.2 kPa, n = 8) glioblastoma xenografts, intracranially propagated luc-MDA-MB-231-LM2-4 (Gd = 3.7 ± 0.2 kPa, Gl = 2.2 ± 0.1 kPa, n = 7) breast cancer xenografts, and Th-MYCN neuroblastomas (Gd = 3.5 ± 0.2 kPa, Gl = 2.3 ± 0.2 kPa, n = 5). Positive correlations between both elasticity (r = 0.72, P < 0.0001) and viscosity (r = 0.78, P < 0.0001) were determined with collagen fraction, but not with cellular or vascular density. Treatment with collagenase significantly reduced Gd (P = 0.002) and Gl (P = 0.0006) in orthotopic breast tumors. Texture analysis of extracted images of picrosirius red staining revealed significant negative correlations of entropy with Gd (r = −0.69, P < 0.0001) and Gl (r = −0.76, P < 0.0001), and positive correlations of fractal dimension with Gd (r = 0.75, P < 0.0001) and Gl (r = 0.78, P < 0.0001). MR elastography can thus provide sensitive imaging biomarkers of tumor collagen deposition and its therapeutic modulation.Significance:MR elastography enables noninvasive detection of tumor stiffness and will aid in the development of ECM-targeting therapies.

Contains Supplementary Methods and Table S1 and S2 which summarize the origin, characteristics, provenance and culture conditions of the cell lines used in this study

Oncogenic BRAF is a critical driver of proliferation and survival and is thus a validated therapeutic target in cancer. We have developed a potent inhibitor, termed 1t (CCT239065), of the mutant protein kinase, V600EBRAF. 1t inhibits signaling downstream of V600EBRAF in cancer cells, blocking DNA synthesis, and inhibiting proliferation. Importantly, we show that 1t is considerably more selective for mutated BRAF cancer cell lines compared with wild-type BRAF lines. The inhibitor is well tolerated in mice and exhibits excellent oral bioavailability (F = 71%). Suppression of V600EBRAF-mediated signaling in human tumor xenografts was observed following oral administration of a single dose of 1t. As expected, the growth rate in vivo of a wild-type BRAF human tumor xenograft model is unaffected by inhibitor 1t. In contrast, 1t elicits significant therapeutic responses in mutant BRAF–driven human melanoma xenografts. Cancer Res; 70(20); 8036–44. ©2010 AACR.

Supplementary Figure 1 from A Novel, Selective, and Efficacious Nanomolar Pyridopyrazinone Inhibitor of V600EBRAF

Supplementary Table 1 from A Novel, Selective, and Efficacious Nanomolar Pyridopyrazinone Inhibitor of V600EBRAF

Supplementary Table 2 from A Novel, Selective, and Efficacious Nanomolar Pyridopyrazinone Inhibitor of V600EBRAF

Scripta materialia 233, 115515 (2023). doi:10.1016/j.scriptamat.2023.115515, Published by Elsevier Science, Amsterdam [u.a.]