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. 2020 Mar 20;10:5178. doi: 10.1038/s41598-020-62088-7

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Delanzomib has in vivo antitumor activity in IM-sensitive and IM-resistant GIST xenografts. (A) Relative tumor volume of UZLX-GIST1 xenografts over the course of a 21-day treatment with vehicle, delanzomib or imatinib. Measurements represent the average of at least four tumors per group. (B) Histopathologic response of imatinib-sensitive UZLX-GIST1 xenografts to treatment with delanzomib in comparison with placebo-treated and imatinib-treated controls. Hematoxylin and eosin (H&E) as well as immunohistochemical staining for Ki-67, phosphorylated histone H3 S10 or cleaved PARP (10X magnification). Histopathologic response grade was determined according to Agaram et al.⁵⁴. Data shown in graph represent at least six tumors per group. Quantification of proliferation index (% Ki-67 positive cells), mitotic cells (pH3-positive cells per 10 high-power fields, HPF) and percentage of cleaved PARP-positive cells (brown staining, respectively) represents the average of at least six tumors per group. It is of note, that only intact, individual cells were included in the counts. Hence, the central debris area in the cleaved PARP stain (although massively positive) is not reflected in the accompanying graph. Columns, mean + SE; *p ≤ 0.05 in comparison to control; **p ≤ 0.01 in comparison to control; ***p ≤ 0.001 in comparison to control (Student’s t-test, 2-tailed). (C) Immunoblot analysis of UZLX-GIST1 xenografts treated with delanzomib (24 h) in comparison with placebo and imatinib-treated positive control. Each lane represents one xenograft (C, vehicle control; DLZ, delanzomib; IM, imatinib). Grouped immunoblot images are either cropped from different parts of the same gel or from a separate gel run with another aliquot of the same protein lysate. (D) Histopathologic response of imatinib-resistant GIST430 xenografts to treatment with delanzomib in comparison with placebo. (E) Immunohistochemical staining for phosphorylated histone H3 S10 in GIST430 xenografts treated with delanzomib or placebo control.