Background Tasquinimod (a quinoline-3-carboxyamide) is a small molecule immunotherapy with demonstrated effects on the tumor microenvironment (TME) involving immunomodulation, anti-angiogenesis and inhibition of metastasis. A target molecule of tasquinimod is the inflammatory protein S100A9 which has been shown to affect the accumulation and function of suppressive myeloid cell subsets in tumors. Given the major impact of myeloid cells to the tumor microenvironment, manipulation of this cell compartment is a desirable goal in cancer therapeutics. Methods To understand the consequences of tasquinimod treatment on the TME, we evaluated early treatment effects in tumor infiltrating myeloid cells. Cellular phenotypes were studied by flow cytometry while gene expression both in tumor tissue and in isolated CD11b⁺ cells or tumor cells were measured by real time-PCR. Effects on angiogenesis were monitored by changes in CD31 levels and by gene expression in tumor tissue. Effects on cytokine levels in tumor tissue and serum were determined by multiplex analysis. Results The MC38-C215 colon carcinoma tumors showed a substantial infiltration of primarily myeloid cells that were dominated by Ly6C^lowF4/80⁺CD206⁺ M2-polarized tumor associated macrophages (TAMs), an immuno-suppressive and pro-angiogenic cell population. Here, we show that tasquinimod treatment induces an anti-tumor effect which is subsequent to a reduction in tumor infiltrating CD206⁺ M2 macrophages and a simultaneous increase in M1 macrophages expressing MHC class II and CD86. The tasquinimod-induced changes in TAM polarization were evident within 24 h of exposure, emphasizing the ability of tasquinimod to rapidly reprogram the tumor microenvironment. This change in the tumor associated myeloid compartment preceded an increased IL12-production within the tumor and a decrease in tumor neovascularization. The switch in TAM polarization by tasquinimod was confirmed in the 4T1 breast cancer model where tasquinimod also reduce lung metastasis development. Conclusion Our data show that tasquinimod affects tumor infiltrating myeloid cells early after exposure, leading to a change in phenotype from pro-angiogenic and immunosuppressive M2-like TAMs to pro-inflammatory M1-like macrophages. These changes are consistent with the effects of tasquinimod seen on tumor vascularization, immune suppression and metastasis giving further insights to the anti-tumor mechanism of action of tasquinimod.

Treatment of neuroendocrine tumours with the radiolabelled somatostatin receptor subtype 2 (SST₂) peptide agonist [¹⁷⁷Lu]Lu-DOTA-TATE is effective and well-established. Recent studies suggest improved therapeutic efficacy using the SST₂ peptide antagonist [¹⁷⁷Lu]Lu-OPS201. However, little is known about the cellular mechanisms that lead to the observed differences. In the present in vitro study, we compared kinetic binding, saturation binding, competition binding, cellular uptake and release of [¹⁷⁷Lu]Lu-OPS201 versus [¹⁷⁷Lu]Lu-DOTA-TATE using HEK cells stably transfected with the human SST₂. While [¹⁷⁷Lu]Lu-OPS201 and [¹⁷⁷Lu]Lu-DOTA-TATE exhibited comparable affinity (K_D, 0.15 ± 0.003 and 0.08 ± 0.02 nM, respectively), [¹⁷⁷Lu]Lu-OPS201 recognized four times more binding sites than [¹⁷⁷Lu]Lu-DOTA-TATE. Competition assays demonstrated that a high concentration of the agonist displaced only 30% of [¹⁷⁷Lu]Lu-OPS201 bound to HEK-SST₂ cell membranes; an indication that the antagonist binds to additional sites that are not recognized by the agonist. [¹⁷⁷Lu]Lu-OPS201 showed faster association and slower dissociation than [¹⁷⁷Lu]Lu-DOTA-TATE. Whereas most of [¹⁷⁷Lu]Lu-OPS201 remained at the cell surface, [¹⁷⁷Lu]Lu-DOTA-TATE was almost completely internalised inside the cell. The present data identified distinct differences between [¹⁷⁷Lu]Lu-OPS201 and [¹⁷⁷Lu]Lu-DOTA-TATE regarding the recognition of receptor binding sites (higher for [¹⁷⁷Lu]Lu-OPS201) and their kinetics (faster association and slower dissociation of [¹⁷⁷Lu]Lu-OPS201) that explain, to a great extent, the improved therapeutic efficacy of [¹⁷⁷Lu]Lu-OPS201 compared to [¹⁷⁷Lu]Lu-DOTA-TATE.

Limited experiments have compared the treatment effects of repetitive cycles of radiolabelled somatostatin (SST) analogues. In vitro and in vivo experiments were conducted in an AR42J cancer cell model, comparing the antagonist [¹⁷⁷Lu]Lu-satoreotide tetraxetan with the agonist [¹⁷⁷Lu]Lu-DOTA-TATE in terms of their binding properties, biodistribution, anti-tumour activity and toxicity. Histopathological and immunohistochemical examinations were performed at different timepoints. In the in vitro assays, [¹⁷⁷Lu]Lu-satoreotide tetraxetan recognised twice as many SST₂ binding sites as [¹⁷⁷Lu]Lu-DOTA-TATE. In mice treated once a week for four consecutive weeks, [¹⁷⁷Lu]Lu-satoreotide tetraxetan (15 MBq) revealed a significantly greater median time taken to reach a tumour volume of 850 mm³ (68 days) compared to [¹⁷⁷Lu]Lu-DOTA-TATE at 15 MBq (43 days) or 30 MBq (48 days). This was associated with a higher tumour uptake, enhanced DNA damage and no or mild effects on body weight, haematological toxicity, or renal toxicity with [¹⁷⁷Lu]Lu-satoreotide tetraxetan (15 MBq). At the end of the study, complete tumour senescence was noted in 20% of animals treated with [¹⁷⁷Lu]Lu-satoreotide tetraxetan, in 13% of those treated with [¹⁷⁷Lu]Lu-DOTA-TATE at 30 MBq, and in none of those treated with [¹⁷⁷Lu]Lu-DOTA-TATE at 15 MBq. In conclusion, repeated administrations of [¹⁷⁷Lu]Lu-satoreotide tetraxetan were able to potentiate peptide receptor radionuclide therapy with a higher tumour uptake, longer median survival, and enhanced DNA damage, with a favourable efficacy/safety profile compared to [¹⁷⁷Lu]Lu-DOTA-TATE.