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Data Supplement from Neuronal Pentraxin 2 Supports Clear Cell Renal Cell Carcinoma by Activating the AMPA-Selective Glutamate Receptor-4

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posted on 2023-03-30, 22:44 authored by Christina A. von Roemeling, Derek C. Radisky, Laura A. Marlow, Simon J. Cooper, Stefan K. Grebe, Panagiotis Z. Anastasiadis, Han W. Tun, John A. Copland

Supplementary Figure 1. (A) QPCR for NPTX2 expression in an array of normal (n=4) and ccRCC (n=7) cell lines. All samples are normalized to a water negative control. Four shRNA lentiviral constructs (sh804, sh855, sh1316, sh1623) against NPTX2 were used to infect A498 cells. (B) Description of ccRCC cell line VHL mutational status. (C) QPCR for NPTX2 mRNA in each of the A498 NT vs. target lentiviral clones. (D) Western blot analysis for NPTX2 protein expression in all four A498 NPTX2 shRNA clones compared to NT control. Protein expression level quantitation is normalized to β-actin. (E) Proliferation (7 day) rescue assay of A498 cells transfected with either a human NPTX2 expression vector (+NPTX2) plasmid or empty vector (EV) control, and infected with NT control, sh1316 or sh1623 NPTX2 targeting lentivirus.

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ARTICLE ABSTRACT

Clear cell renal cell carcinoma (ccRCC) is the most common subtype of kidney cancer and has the highest propensity to manifest as metastatic disease. Recent characterizations of the genetic signature of ccRCC have revealed several factors correlated with tumor cell migration and invasion; however, the specific events driving malignancy are not well defined. Furthermore, there remains a lack of targeted therapies that result in long-term, sustainable response in patients with metastatic disease. We show here that neuronal pentraxin 2 (NPTX2) is overexpressed specifically in ccRCC primary tumors and metastases, and that it contributes to tumor cell viability and promotes cell migration through its interaction with the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor subunit GluR4. We propose NPTX2 as a novel molecular target for therapy for patients with ccRCC diagnosed with or at risk of developing metastatic disease. Cancer Res; 74(17); 4796–810. ©2014 AACR.