Abstract 3017: Lack of MCPIP1 enzymatic activity leads to neoplastic transformation of normal kidney cells
Authors: Marona Paulina , Piasecka Iga , Gorka Judyta , Kwapisz Oliwia , Myrczek Rafal , Pospiech Ewelina , Jura Jolanta , Miekus Katarzyna 
Published: March 2024

Introduction: The MCPIP1 protein (Monocyte Chemotactic Protein-1 Induced Protein) is involved in the negative regulation of inflammation due to its RNase activity. A growing number of publications suggest that the MCPIP1 protein may influence the development of cancer by regulation of factors involved in angiogenesis, proliferation and cell death. We believe, that MCPIP1 may be a potential tumor suppressor. Our recent results show that lack of enzymatic activity of MCPIP1 leads to increased tumor growth, angiogenesis and metastasis. The main aim of our research is to study the importance of MCPIP1 for neoplastic transformation of normal, epithelial kidney cells, as well as acquirement of stemness features.

Material and Methods: To examine the effect of MCPIP1 in normal epithelial cell lines TCMK-1, RPTEC-Tert1 and HK-2 were transduced to overexpress mutated, inactive form of MCPIP1 (pLIX D141N) with proper control (pLIX PURO). We analyzed cells clonogenicity, as well as the levels of CSCs markers by western blot, NGS and qPCR. Next, cells were injected subcutaneously into NOD-SCID mice to check if mutation of MCPIP1 will induce tumor growth in vivo. Mice blood was analyzed for the presence of circulating tumor cells (CTCs), and secretion of cytokines by Proteome Profiler assay. Tumors were evaluated by multiple IHC stainings, as well as western blot and qPCR.

Results and Discussion: We have shown that the mutation of MCPIP1 increases the clonogenicity of kidney cells. Moreover, pLIX D141N cells were characterized by higher levels of the c-Met receptor, vimentin, Twist, CD44 and c-Myc, which are the markers of epithelial to mesenchymal transition and cancer stemness. The obtained results suggest that the loss of the MCPIP1 protein causes the acquisition of the characteristics of neoplastic cells. Next, we checked whether MCPIP1 mutation in normal cells predisposes them to proliferate when administered to mice. We found that, D141N mutation caused the growth of large tumors with higher expression of Cd44, Klf4 and Myc and number of CTCs, while the control cells injected into mice developed only local fibrosis with high expression of aSMA. pLIX D141N tumors secreted CXCL16 into mice plasma, and were characterized by increased phosphorylation of c-Met, Src kinase and c-Myc.

Conclusions: We believe that the MCPIP1 protein may be a marker of tumor initiation and play a key role in neoplastic transformation by regulating the changes in cell phenotype and levels of CSCs markers.

This study was supported by National Science Center grants no. 2022/47/B/NZ5/02724, 2022/45/B/NZ5/01973 and MNS 15/2021.

Citation Format: Paulina Marona, Iga Piasecka, Judyta Gorka, Oliwia Kwapisz, Rafal Myrczek, Ewelina Pospiech, Jolanta Jura, Katarzyna Miekus. Lack of MCPIP1 enzymatic activity leads to neoplastic transformation of normal kidney cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3017.