Abstract 2377: microRNA expression profile in urinary exosomes is dependent on non-invasive lymphoma induction in mice

B. Wilson, Rebekah Betar, Alexander Martin, Z. Niazi, Michael P. Boyer, Lori Winter, V. Babich, F. Sole, E. Ananieva
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Abstract

Lymphoma accounts for approximately 4% of cancers in the United States, with an estimated 20,910 number of deaths per year. The standard method of diagnosis and staging of lymphoma involves surgical biopsy of the tumor - a procedure that has many negative associated risks. Exosomes are extracellular vesicles secreted in biological fluids that can serve as “liquid biomarkers”. Identification of unique cancer-related biomarkers in urinary exosomes may provide a novel non-invasive and cost-effective tool for lymphoma diagnosis. Analyses of biomarkers obtained from urinary exosomes have been used to evaluate urological cancers, however, these methods have not yet been translated to non-urological pathologies, such as lymphomas. The objective for this study was to determine the profile of microRNAs (miRNAs) expressed in urinary exosomes of mice challenged with lymphoma and compare it to miRNAs identified in urinary exosomes of control mice. Male and female C57BL/6 mice, (tumor (+), n=12), were injected with either 2.5x105 mouse EL-4 lymphoma cells or phosphate-buffered saline (tumor (-), n=12). Tumor growth was monitored for 20 days. Urine was collected for 48 hours starting on day 17 and serum, tumor tissues, and organs were collected on day 20. Extraction of urinary exosomes was followed by total RNA isolation and RT-qPCR for which a set of PCR arrays consisting of 709 mouse-specific miRNA primers was used. Fold changes in miRNA expression were quantified using the ΔΔCt method. Mice developed tumors by day 13 with initial tumor appearance around day seven. There were no statistically significant differences between final tumor mass, body weight, or food and water intake in tumor (+) versus tumor (-) mice. RT-qPCR arrays of miRNAs extracted from urinary exosomes revealed 464 miRNAs that were differentially expressed between tumor (+) and tumor (-). 215 miRNAs were up-regulated, while 249 miRNAs were down-regulated in tumor (+) mice. These results will be compared to miRNAs from serum and tumor tissues to identify tumor-specific miRNAs that can be used for potential application in the clinical setting. Citation Format: Brittany Wilson, Rebekah Betar, Alexander Martin, Zackaria Niazi, Michael Boyer, Lori Winter, Victor Babich, Francesca Di Sole, Elitsa Ananieva. microRNA expression profile in urinary exosomes is dependent on non-invasive lymphoma induction in mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2377.
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摘要2377:小鼠尿外泌体中的microRNA表达谱依赖于非侵袭性淋巴瘤诱导
在美国,淋巴瘤约占癌症的4%,估计每年有20910人死亡。淋巴瘤诊断和分期的标准方法包括肿瘤的手术活检,这一过程有许多负面的相关风险。外泌体是生物体液中分泌的细胞外囊泡,可作为“液体生物标志物”。识别尿外泌体中独特的癌症相关生物标志物可能为淋巴瘤诊断提供一种新的非侵入性和经济有效的工具。从尿外泌体获得的生物标志物分析已被用于评估泌尿系统癌症,然而,这些方法尚未被转化为非泌尿系统疾病,如淋巴瘤。本研究的目的是确定淋巴瘤小鼠尿外泌体中表达的microRNAs (miRNAs)的谱,并将其与对照小鼠尿外泌体中鉴定的miRNAs进行比较。将雄性和雌性C57BL/6小鼠(肿瘤(+),n=12)分别注射2.5 × 105小鼠EL-4淋巴瘤细胞或磷酸盐缓冲盐水(肿瘤(-),n=12)。监测肿瘤生长20 d。从第17天开始收集尿液48小时,第20天收集血清、肿瘤组织和器官。提取尿外泌体后进行总RNA分离和RT-qPCR,其中使用了一组由709个小鼠特异性miRNA引物组成的PCR阵列。使用ΔΔCt方法定量miRNA表达的折叠变化。小鼠在第13天出现肿瘤,第7天左右出现肿瘤。肿瘤(+)和肿瘤(-)小鼠的最终肿瘤质量、体重、食物和水的摄入量没有统计学上的显著差异。从尿外泌体中提取的mirna的RT-qPCR阵列显示464个mirna在肿瘤(+)和肿瘤(-)之间差异表达。在肿瘤(+)小鼠中,215个mirna上调,249个mirna下调。这些结果将与来自血清和肿瘤组织的mirna进行比较,以确定可用于临床环境潜在应用的肿瘤特异性mirna。引文格式:Brittany Wilson, Rebekah Betar, Alexander Martin, Zackaria Niazi, Michael Boyer, Lori Winter, Victor Babich, Francesca Di Sole, Elitsa Ananieva。小鼠尿外泌体中的microRNA表达谱依赖于非侵袭性淋巴瘤诱导[摘要]。见:美国癌症研究协会2021年年会论文集;2021年4月10日至15日和5月17日至21日。费城(PA): AACR;癌症杂志,2021;81(13 -增刊):摘要nr 2377。
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