Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Oct;11(10):e533.
doi: 10.1002/ctm2.533.

The oral KIF11 inhibitor 4SC-205 exhibits antitumor activity and potentiates standard and targeted therapies in primary and metastatic neuroblastoma models

Marc Masanas  1 , Nuria Masiá  2 , Leticia Suárez-Cabrera  2 , Mireia Olivan  2   3 , Aroa Soriano  1 , Blanca Majem  2 , Laura Devis-Jauregui  4 , Rebeca Burgos-Panadero  5   6 , Carlos Jiménez  1 , Pau Rodriguez-Sodupe  7 , Ariadna Boloix  1 , Ignasi Toledano  2 , Gabriela Guillén  1   8 , Alexandra Navarro  9 , David Llobet-Navas  4   6 , Alberto Villanueva  10   11 , Josep Sánchez de Toledo  1   12 , Josep Roma  1 , Rosa Noguera  5   6 , Lucas Moreno  1   13 , Rolf Krauss  14 , Soledad Gallego  1   13 , Anna Santamaria  2 , Miguel F Segura  1
Affiliations

The oral KIF11 inhibitor 4SC-205 exhibits antitumor activity and potentiates standard and targeted therapies in primary and metastatic neuroblastoma models

Marc Masanas et al. Clin Transl Med. 2021 Oct.
No abstract available

PubMed Disclaimer

Conflict of interest statement

Dr. Moreno participates in data monitoring committees of clinical trials sponsored by Novartis, Actuate Therapeutics, Shionogi, Incyte, the University of Southampton and the Royal Marsden NHS Foundation Trust; and had a consulting role for Novartis and Shionogi. Dr. Lucas Moreno is also a member of the Executive Committee of the European neuroblastoma research cooperative group (SIOPEN) which receives royalties for the sales of dinutuximab beta. Rolf Krauss (RK) is an employee of 4SC. Alberto Villanueva (AV) is co‐founder of Xenopat S.L. No potential conflict of interest was disclosed by the rest of the authors.

Figures

FIGURE 1
FIGURE 1
KIF11 expression is an independent prognostic factor of survival in neuroblastoma. (A) KIF11 mRNA expression levels comparing low‐/intermediate‐ with high‐risk neuroblastoma tumors (GSE62564, n = 498). (B‐D) Kaplan–Meier overall survival curve in a cohort of 498 patients based on KIF11 mRNA expression (B) or stratified in low‐ and intermediate‐risk (C) or high‐risk (D) neuroblastoma subcohorts. (E) KIF11 mRNA expression in neuroblastoma patients with different genomic alterations (GSE3960, n = 101). (F) Representative images of KIF11 immunohistochemistry in low‐and high‐risk neuroblastoma tissues. Scale bar indicates 50 μm. (G and H) Kaplan–Meier curves of event‐free survival (G) and overall survival (H) based on KIF11 protein expression
FIGURE 2
FIGURE 2
Genetic and pharmacological inhibition of KIF11 reduces tumor growth in subcutaneous neuroblastoma xenografts. (A) Analysis of tumor volume of SK‐N‐BE(2) cells transduced with an inducible shKIF11 lentiviral construct comparing the effects of KIF11 silencing (+Dox) versus control (‐Dox). (B) Waterfall plot comparing the change in tumor volume at day 28 post‐injection versus day 18, when doxycycline was added into the drinking water. (C) Western blot analysis of excised tumors at termination of the experiment. Turbo‐RFP (tRFP) reporter expression was used as a control for shRNA transgene induction. (D) 4SC‐205 chemical structure. (E) Dose‐response curves of neuroblastoma cell lines treated with increasing concentrations of 4SC‐205 for 48 h. IC50 values are represented as the average of three independent experiments ± SEM. (F) Mitotic spindle immunofluorescence of SK‐N‐BE(2) cells transfected with siKIF11 or treated with 4SC‐205 (25 nM) for 24 h. KIF11: red, α‐TUBULIN: green, DAPI: blue. Scale bar, 5 μm. (G) Individual tumor growth of xenografts derived from SK‐N‐BE(2) comparing vehicle (n = 10) versus 40 mg/kg 4SC‐205 (n = 10). (H) Waterfall plot comparing the change in tumor volume at day 16 post‐treatment versus day 4. (I) Western blot analysis of cell‐cycle and apoptosis‐related proteins in SK‐N‐BE(2) resected tumors. (J) Tumor growth of subcutaneous xenograft derived from SK‐N‐AS treated with vehicle (n = 10) or 40 mg/kg 4SC‐205 (n = 10). (K) Tumor volume fold change at day 7 post‐treatment versus day 0. (L) Western blot analysis of resected tumors at the end of the experiment. (M) Gene set enrichment analysis of SK‐N‐BE(2) xenografts treated with vehicle or 4SC‐205. Graph represents normalized enrichment score (NES) values of enriched sets with p < 0.05. (N and O) Heatmap representing top 20 differentially deregulated genes of G2/M checkpoint and E2F targets (N), and mTORC signaling‐related genes (O)
FIGURE 3
FIGURE 3
4SC‐205 impairs PDOX growth and prolongs the survival of mice bearing neuroblastoma liver metastasis. (A) Schematic representation of PDOX generation and characterization. (B) Immunohistochemistry of neuroblastoma markers in FFPE tumor sections from the original tumor (upper panels) and after implantation in mice (lower panels). H&E: Hematoxylin and eosin staining. Scale bar represents 110 μm. (C and D) Chromosomal copy number variations (CNV) of the original tumor and after implantation in mice. The most relevant molecular pathogenic alterations found in the original tumor and PDOX are highlighted. (E) Schematic illustration of the treatment schedule. Mice bearing PDOX were treated for 3 weeks with either vehicle (n = 7) or 4SC‐205 (40 mg/kg, n = 11). (F) Tumor weights at the end of the experiment. (G) Representative picture of the dissected tumors (T: tumor; K: kidney). Scale bar: 1 cm. (H) Representative images of phosphorylated histone H3. Scale bar: 10 μm. (I). Quantification of phospho‐histone H3 positive cells in histological sections from vehicle‐ and 4SC‐205‐treated tumors. Graph represents the average percentage of positive cells ± SEM from vehicle‐or 4SC‐205 (10 representative fields/tumor) ‐treated tumors (n = 5/group). (J) Scheme of the experimental design. SK‐N‐BE(2) cells were injected into the tail vein, and 21 days later, mice were randomized into vehicle (n = 13) and 40 mg/kg 4SC‐205 groups (n = 15). The mice received oral administration of 4SC‐205 three times a week for five consecutive weeks. (K) Scatter dot plots representing the average quantification of tumor bioluminescence ± SEM at the indicated days post‐treatment. **p < 0.01, two‐tailed student's t‐test. (L) Representative images of luciferase activity in eight mice from the vehicle and 4SC‐205 treatment groups at 28 days posttreatment. (M) Kaplan–Meier survival curve of mice with neuroblastoma liver metastases treated with either vehicle or 40 mg/kg 4SC‐205 for 5 weeks. Statistical differences were calculated using the Gehan–Breslow–Wilcoxon test. *p < 0.05, ***p < 0.001, two‐tailed Student's t‐test
FIGURE 4
FIGURE 4
4SC‐205 potentiates the effect of chemotherapy and neuroblastoma‐targeted therapies. (A) Scheme of the experimental design for the combination of 4SC‐205 and standard chemotherapies. Images are representative of crystal violet staining of SK‐N‐BE(2) cells treated with CDDP (1000 nM), doxorubicin (15 nM), topotecan (10 nM), 4SC‐205 (17.5 nM) and their corresponding combinations (Combo). (B) Heatmaps showing the percentage of cellular fraction affected by drug combination treatments. (C) Graphs represent the average effect on cell viability from three independent experiments ± SEM (n = 3/condition). (D) Combinatorial analysis performed using SynergyFinder 2.0 software. (E) Schematic representation of the experimental design combining 4SC‐205 and neuroblastoma‐targeted therapies. Representative crystal violet staining images of the ALK‐mutated SH‐SY5Y cell line treated with the ALK inhibitors ceritinib (62.5 nM) or lorlatinib (50 nM) with or without 4SC‐205 (15 nM). (F) Heatmaps show the percentage of the cellular fraction affected by the drug combination treatments at the indicated doses. The graph represents the average of three independent experiments (n = 3/condition) ± SEM at 62.5 nM and 50 nM of ceritinib or lorlatinib, respectively. (G) Combinatorial analysis of 4SC‐205 and ceritinib/lorlatinib in SH‐SY5Y and KELLY cells. (H) Crystal violet staining images of SK‐N‐BE(2) cells treated with selumetinib (250 nM) and 4SC‐205 (20 nM) and their respective combinations. (I) Heatmap showing the fraction of cells affected after the combination of 4SC‐205 and selumetinib at the indicated concentrations for 48 h. Graph represents the average percentage of cell proliferation (n = 3/condition) ± SEM at 250 nM of selumetinib. (J) Combinatorial analysis of 4SC‐205 and selumetinib in SK‐N‐BE(2) and SK‐N‐AS. *p < 0.05, **p < 0.01, ***p < 0.001, two tailed Student's t‐test
See this image and copyright information in PMC

References

    1. Matthay KK, Maris JM, Schleiermacher G, et al. Neuroblastoma. Nat Rev Dis Primers. 2016;2:16078. - PubMed
    1. Stafman LL, Beierle EA. Cell proliferation in neuroblastoma. Cancers. 2016;8:13. - PMC - PubMed
    1. London WB, Shimada H, d'Amore E, et al. Age, tumor grade, and mitosis‐karyorrhexis index (MKI) are independently predictive of outcome in neuroblastoma (NB). J Clin Oncol. 2007;25(18_suppl):9558.
    1. Blangy A, Lane HA, d'Herin P, Harper M, Kress M, Nigg EA. Phosphorylation by p34cdc2 regulates spindle association of human Eg5, a kinesin‐related motor essential for bipolar spindle formation in vivo. Cell. 1995;83(7):1159–1169. - PubMed
    1. Hansson K, Radke K, Aaltonen K, et al. Therapeutic targeting of KSP in preclinical models of high‐risk neuroblastoma. Sci Transl Med. 2020;12(562):eaba4434. - PubMed

Publication types

LinkOut - more resources