Herein, we demonstrated that MDK serum levels increased in patients with SCLC, and MDK regulated cell proliferation and the antitumor effect of CDDP in vitro and in vivo. To the best of our knowledge, this is the first study to evaluate the expression and functions of MDK in SCLC.
Several clinical trials on targeted therapies for SCLC have been performed and numerous are ongoing; however, the reported data are unsatisfactory [33]. SCLC is heterogeneous, and various SCLC subtypes exist simultaneously within a tumor, indicating that comprehensive treatment, and not just a single treatment, is necessary for the cancer [5, 10, 11, 34]. Lim et al. proposed that non-NE-type SCLC cells secrete MDK, which promotes the growth of NE-type SCLC cells [10]. Thus, we hypothesized that MDK-targeted therapy might be sufficient for treating tumors with various SCLC cells because MDK might target non-NE-type SCLC cells, thereby indirectly inhibiting the proliferation of NE-type SCLC cells. Although the present results did not show a correlation between SCLC subtypes or NE features and MDK expression in human tumor tissues and SCLC cell lines, MDK was expressed regardless of the subtype, and both non-NE- and NE-type SCLC cells were sensitive to MDK inhibition. These results suggest that MDK may be a versatile therapeutic target for SCLC. Nevertheless, further studies are required to clarify the association between MDK expression and SCLC characteristics.
Upregulated MDK expression in tumor tissues is associated with tumor growth in many cancers, including non-SCLC (NSCLC), hepatocellular carcinoma, and bladder cancer [17, 24, 25]. Furthermore, compared with healthy controls, patients with several cancers exhibit upregulated s-MDK expression, which is useful as a non-invasive biomarker for diagnosis and disease progression [17, 35–39]. Herein, we found increased s-MDK levels in patients with SCLC for the first time and observed a correlation between increased s-MDK levels and tumor burden in SCLC. Mostly, SCLC is diagnosed at an advanced stage, and owing to its high malignant potential, prompt treatment is often required. Thus, determining s-MDK levels can be helpful for the clinical diagnosis of SCLC. Furthermore, we found that s-MDK levels decreased with treatment in patients with high s-MDK secretion, and in one case (Fig. S1a), increased s-MDK levels were detected before disease progression based on imaging and tumor markers, indicating that it might be a predictive factor for disease progression during treatment.
MDK is involved in cancer cell proliferation, survival, migration, and resistance to chemotherapy via the PI3K/AKT and MAPK pathways [27, 40, 41]. Herein, MDK activated the AKT pathway in SCLC; however, its correlation to the MAPK pathway remained unclear. Hao et al. reported that MDK inhibition by iMDK suppressed only the PI3K/AKT pathway in NSCLC; however, the reason why the MAPK pathway was not inhibited remained unclear [25]. In our study, P-ERK was upregulated in H69 after MDK suppression, which might be compensated for by the downregulation of PI3K/AKT pathway. In MS1L cells, both the PI3K/AKT and MAPK pathways were enhanced after MDK overexpression, which indicated that pathway inhibition or activation might be cell-context-dependent.
We observed the combined antitumor effects of iMDK and CDDP in vitro and in vivo. Moreover, MDK and the PI3K/AKT pathway were upregulated in the CDDP-resistant SCLC cell lines. CDDP is one of the chemotherapeutic drugs currently used for SCLC; however, acquired resistance to this drug affects treatment efficacy. MDK is overexpressed in several cancers and mediates resistance to chemotherapeutic drugs via several mechanism [27]. MDK renders glioma cells resistant to tetrahydrocannabinol and inhibits autophagy-mediated cell death activation via the AKTmTORC1 pathway [42]. In gastric cancer, MDK overexpression upregulates P-AKT and P-ERK expression, which induces resistance to adriamycin [43]. Ying et al. reported that PI3K/AKT pathway activation was associated with resistance to chemotherapy in SCLC [32], indicating that PI3K/AKT pathway suppression might be the main reason why MDK inhibition in the present study enhanced the efficacy of CDDP.
Nevertheless, the present study had several limitations. First, owing to the small number of IHC evaluations and SCLC cell lines, concluding that no correlation exists between MDK expression and SCLC subtypes is difficult. Second, the age of control population for s-MDK measurement was relatively less than that of patients with SCLC included in this study. Reportedly, s-MDK gradually decreases with age in healthy children; hence, the difference in age may have had affected the results [44]. Third, owing to the small sample size and variable disease stages of patients whose MDK was measured, evaluating the correlation between s-MDK and prognosis was difficult. Lastly, although ICI combined with chemotherapy has become one of the standard therapies for SCLC, we did not evaluate the effect of MDK on the effects of ICIs. MDK suppressed CD8 T-cell functions by stimulating immunosuppressive myeloid-derived suppressor cells, which was driven in part by nuclear factor-κB in the TME, which resulted in resistance to ICI [22, 23]. If MDK inhibition improves T-cell functions and TME in SCLC, the combination of iMDK with ICI combined with platinum-based chemotherapy might be promising.
Altogether, this is the first study to evaluate the expression and functions of MDK in SCLC, and the finding that MDK inhibition enhances the efficacy of CDDP is crucial because it presents MDK as a potential therapeutic target for SCLC. The present findings show the pivotal role of MDK in SCLC, and thus, suggest MDK as a potential therapeutic target for SCLC treatment.