Elevated TIMP-1 expression is associated with a prometastatic phenotype, disease relapse, and poor survival in neuroblastoma

Abstract

Approximately two-thirds of patients with neuroblastoma are found to have metastatic disease at time of diagnosis with frequent skeletal, lymph node, central nervous system, and liver involvement. Using a serial in vivo splenic injection model, we have isolated an aggressive subclone (BE(2)-C/LM2) from MYCN-amplified neuroblastomas that demonstrate an enhanced propensity to develop metastatic liver lesions. BE(2)-C/LM2 subclone cells demonstrate increased adherent, soft agar colony and tumorsphere growth in vitro. Components of the tumor microenvironment regulate cancer progression, via networks of cytokines and growth factors. Cytokine array analysis identified increased TIMP-1 in the plasma of mice injected with BE(2)-C/LM2 subclone cells, leading us to hypothesize that TIMP-1 may play a role in our observed prometastatic phenotype. Immunoblotting and ELISA demonstrated enhanced endogenous TIMP-1 expression in our isolated neuroblastoma subclone. Silencing endogenous TIMP-1 successfully blocked in vitro proliferation, soft agar colony formation and tumorsphere formation by BE(2)-C/LM2 cells. Stable RNA interference of endogenous TIMP-1 failed to reverse the prometastatic phenotype of our BE(2)-C/LM2 subclone in our liver metastasis model, suggesting that endogenous TIMP-1 levels may not be an essential component of this in vivo behavior. Notably, tissue microarray analysis and Kaplan-Meier by gene expression demonstrates that elevated TIMP-1 expression is correlated with increased disease relapse and mortality in patients with neuroblastoma. Taken together, our study identifies TIMP-1 as a novel soluble factor that is associated with a prometastatic phenotype in our in vivo model and adverse outcomes in patients with neuroblastoma.

Nanoformulation of CCL21 greatly increases its effectiveness as an immunotherapy for neuroblastoma

“…However, unexpectedly, nanoformulated CCL21 also increased the presence of TIMP-1 (Fig. 13), which has been reported to have pro-tumor effects in the neuroblastoma setting [85].…”

Induction of serine hydroxymethyltransferase 2 promotes tumorigenesis and metastasis in neuroblastoma

“…To further evaluate the role of SHMT2 on NB metastatic potential, the LM2 cell line was evaluated. The LM2 cell line, which was established in our laboratory previously [ 13 ], is a highly aggressive NB cell line with propensity to metastasize that was obtained from BE(2)-C cell liver metastases after splenic injection for two cycles. In order to assess for SHMT2 mRNA and protein expression, RT-qPCR and immunoblotting were performed.…”

DEAD-Box Helicase 4 (Ddx4)+ Stem Cells Sustain Tumor Progression in Non-Serous Ovarian Cancers

“…In particular, upregulated DEGs included SIRT1 , which is involved in stemness [ 28 ] and chemoresistance [ 29 ], SNAIL2, which participates in EMT and collagen remodeling [ 30 , 31 ], and SIRT2 , CTGF , IL-11 and MMP13 , involved in cell migration, invasiveness as well as resistance to platinum and paclitaxel [ 32 , 33 , 34 , 35 , 36 ]. Other upregulated DEGs, such as MCM-6 , TGFB-1 , HMGA2 and FOXM1 , are involved in similar cancer-related processes [ 37 , 38 , 39 , 40 , 41 , 42 ], while downregulated DEGs, such as LOX and TIMP-1 (which are associated with poor cancer prognosis [ 43 , 44 , 45 ]), were observed in A2780 co-cultured with both A1 and A2 derived ML-Ddx4 + cells. Among these gene expression alterations, only ADGRL2 and PES1 upregulation were observed in A2780 cells conditioned with control ML-Ddx4 + cells, in a similar fashion to what emerged from the A1-related co-culture.…”