Molecular imaging of EGFR and CD44v6 for prediction and response monitoring of HSP90 inhibition in an in vivo squamous cell carcinoma model

Spiegelberg, Diana; Mortensen, Anja C.; Selvaraju, Ram Kumar; Eriksson, Olof; Stenerlöw, Bo; Nestor, Marika

doi:10.1007/s00259-015-3260-x

Cited by 23 publications

(23 citation statements)

References 23 publications

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“…The higher CD44v6-density on HCT116 cells was in line with results from LigandTracer analyses, where a clear signal was detected from HCT116 cells, whereas the signal from UM-SCC-74B cells was below the detection limit of the LigandTracer instrument. This supports previous studies assessing the CD44v6-antigen expression of these cell lines, verifying that CD44v6-antigen density on HCT116 and UM-SCC-74B cells can be considered moderate and low, respectively [27][28][29].…”

Section: Discussionsupporting

confidence: 91%

Enhancing the therapeutic effects of in vitro targeted radionuclide therapy of 3D multicellular tumor spheroids using the novel stapled MDM2/X-p53 antagonist PM2

et al. 2020

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Background: Precision therapeutics continuously make advances in cancer therapy, and a field of growing interest is the combination of targeted radionuclide therapy (TRNT) with potential radiosensitizing agents. This study evaluated whether the effects of in vitro TRNT, using the 177 Lu-labeled anti-CD44v6 antibody AbN44v6, were potentiated by the novel stapled MDM2/X-p53 antagonist PM2. Materials and methods: Two wt p53 cell lines, HCT116 (colorectal carcinoma) and UM-SCC-74B (head and neck squamous cell carcinoma), expressing different levels of the target antigen, CD44v6, were used. Antigen-specific binding of 177 Lu-AbN44v6 was initially verified in a 2D cell assay, after which the potential effects of unlabeled AbN44v6 on downstream phosphorylation of Erk1/2 were evaluated by western blotting. Further, the therapeutic effects of unlabeled AbN44v6, 177 Lu-AbN44v6, PM2, or a combination (labeled/unlabeled AbN44v6 +/− PM2) were assessed in 3D multicellular tumor spheroid assays. Results: Radiolabeled antibody bound specifically to CD44v6 on both cell lines. Unlabeled AbN44v6 binding did not induce downstream phosphorylation of Erk1/2 at any of the concentrations tested, and repeated treatments with the unlabeled antibody did not result in any spheroid growth inhibition. 177 Lu-AbN44v6 impaired spheroid growth in a dose-dependent and antigen-dependent manner. A single modality treatment with 20 μM of PM2 significantly impaired spheroid growth in both spheroid models. Furthermore, the combination of TRNT and PM2based therapy proved significantly more potent than either monotherapy. In HCT116 spheroids, this resulted in a two-and threefold spheroid growth rate decrease for the combination of PM2 and 100 kBq 177 Lu-AbN44v6 compared to monotherapies 14-day post treatment. In UM-SCC-74B spheroids, the combination therapy resulted in a reduction in spheroid size compared to the initial spheroid size 10-day post treatment. Conclusion: TRNT using 177 Lu-AbN44v6 proved efficient in stalling spheroid growth in a dose-dependent and antigen-dependent manner, and PM2 treatment demonstrated a growth inhibitory effect as a monotherapy. Moreover, by combining TRNT with PM2-based therapy, therapeutic effects of TRNT were potentiated in a 3D multicellular tumor spheroid model. This proof-of-concept study exemplifies the strength and possibility of combining TRNT targeting CD44v6 with PM2-based therapy.

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Section: Discussionsupporting

confidence: 91%

Enhancing the therapeutic effects of in vitro targeted radionuclide therapy of 3D multicellular tumor spheroids using the novel stapled MDM2/X-p53 antagonist PM2

et al. 2020

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“…The receptor expression was reduced by more than 50% in the named groups. The downregulation of HSP90 as well as its client proteins is a very attractive measure for treatment efficacy in vivo and expression levels of EGFR or HER2 among others have been assessed for treatment response monitoring with molecular imaging techniques like PET and SPECT cameras [14][15][16] .…”

Section: Discussionmentioning

confidence: 99%

The HSP90 inhibitor Onalespib exerts synergistic anti-cancer effects when combined with radiotherapy: an in vitro and in vivo approach

Spiegelberg

Abramenkovs

Mortensen

et al. 2020

Sci Rep

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Oncogenic client-proteins of the chaperone Heat shock protein 90 (HSP90) insure unlimited tumor growth and are involved in resistance to chemo-and radiotherapy. The HSP90 inhibitor Onalespib initiates the degradation of oncoproteins, and might also act as a radiosensitizer. The aim of this study was therefore to evaluate the efficacy of Onalespib in combination with external beam radiotherapy in an in vitro and in vivo approach. Onalespib downregulated client proteins, lead to increased apoptosis and caused DNA-double-strands. Monotherapy and combination with radiotherapy reduced colony formation, proliferation and migration assessed in radiosensitive HCT116 and radioresistant A431 cells. In vivo, a minimal treatment regimen for 3 consecutive days of Onalespib (3 × 10 mg/kg) doubled survival, whereas Onalespib with radiotherapy (3 × 2 Gy) caused a substantial delay in tumor growth and prolonged the survival by a factor of 3 compared to the HCT116 xenografted control group. Our results demonstrate that Onalespib exerts synergistic anti-cancer effects when combined with radiotherapy, most prominent in the radiosensitive cell models. We speculate that the depletion and downregulation of client proteins involved in signalling, migration and DNA repair mechanisms is the cause. Thus, individually, or in combination with radiotherapy Onalespib inhibits tumor growth and has the potential to improve radiotherapy outcomes, prolonging the overall survival of cancer patients.

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“…As Onalespib can influence angiogenesis and the expression of numerous client proteins [32,33], the binding and spatial distribution of 177 Lu-DOTATATE after Onalespib treatment as well as expression of SSTR2 was investigated. Ex vivo autoradiography of BON tumor sections 48 h after the last treatment revealed that Onalespib treatment did not alter the uptake of 177 Lu-DOTATATE in the assessed sections (Fig.…”

Section: Lu-dotatate Selectivity and Distributionmentioning

confidence: 99%

“…EGFR and VEGFR are HSP90 client proteins and can be used to assess molecular responses after Onalespib therapy [15,30,33]. Here, the expression of EGFR and VEGFR was assessed in tumors with immunohistochemistry 48 h after the last treatment ( Fig.…”

Section: Molecular Response To Treatmentmentioning

confidence: 99%

The radiosensitizer Onalespib increases complete remission in 177Lu-DOTATATE-treated mice bearing neuroendocrine tumor xenografts

Lindström

Spiegelberg

Raval

et al. 2020

Eur J Nucl Med Mol Imaging

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Purpose 177 Lu-DOTATATE targeting the somatostatin receptor (SSTR) is utilized for treatment of neuroendocrine tumors (NETs). Onalespib, a heat shock protein 90 (HSP90) inhibitor, has demonstrated radiosensitizing properties and may thus enhance the effect of 177 Lu-DOTATATE. Consequently, the aim of this study was to assess the potential of Onalespib in combination with 177 Lu-DOTATATE in vivo and to examine the toxicity profiles of the treatments. Methods 177 Lu-DOTATATE selectivity and distribution in NET xenografts were studied using biodistribution and autoradiography. Therapeutic effects of Onalespib in combination with 177 Lu-DOTATATE were studied in NET xenografts. Histological analyses were used to assess molecular effects from treatment and to establish toxicity profiles. Results Biodistribution and autoradiography confirmed the SSTR-selective tumor uptake of 177 Lu-DOTATATE, which was unaffected by Onalespib treatment. Immunohistochemistry verified molecular responses to Onalespib therapy in the tumors. While Onalespib and 177 Lu-DOTATATE monotherapies resulted in a 10% and 33% delay in tumor doubling time compared with control, the combination treatment resulted in a 73% delayed tumor doubling time. Moreover, combination treatment increased complete remissions threefold from 177 Lu-DOTATATE monotherapy, resulting in 29% complete remissions. In addition, histological analyses demonstrated radiation-induced glomerular injury in the 177 Lu-DOTATATE monotherapy group. The damage was decreased tenfold in the combination group, potentially due to Onalespib-induced HSP70 upregulation in the kidneys. Conclusion Treatment with Onalespib potentiated 177 Lu-DOTATATE therapy of NET xenografts with a favorable toxicity profile. Utilizing Onalespib's radiosensitizing properties with 177 Lu-DOTATATE may lead to better therapeutic results in the future and may reduce unwanted side effects in dose-limiting organs.

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Molecular imaging of EGFR and CD44v6 for prediction and response monitoring of HSP90 inhibition in an in vivo squamous cell carcinoma model

Cited by 23 publications

References 23 publications

Enhancing the therapeutic effects of in vitro targeted radionuclide therapy of 3D multicellular tumor spheroids using the novel stapled MDM2/X-p53 antagonist PM2

Enhancing the therapeutic effects of in vitro targeted radionuclide therapy of 3D multicellular tumor spheroids using the novel stapled MDM2/X-p53 antagonist PM2

The HSP90 inhibitor Onalespib exerts synergistic anti-cancer effects when combined with radiotherapy: an in vitro and in vivo approach

The radiosensitizer Onalespib increases complete remission in 177Lu-DOTATATE-treated mice bearing neuroendocrine tumor xenografts

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