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血清细胞外囊泡表达前列腺特异性膜抗原在泌尿系统恶性肿瘤中的诊断潜力。

Diagnostic potential of serum extracellular vesicles expressing prostate-specific membrane antigen in urologic malignancies.

机构信息

Research Team for Mechanism of Aging, Tokyo Metropolitan Institute of Gerontology, 35-2 Sakae-cho, Itabashi-ku, Tokyo, 173-0015, Japan.

Biological Process of Aging, Tokyo Metropolitan Institute of Gerontology, 35-2 Sakae-cho, Itabashi-ku, Tokyo, 173-0015, Japan.

出版信息

Sci Rep. 2021 Jul 22;11(1):15000. doi: 10.1038/s41598-021-94603-9.

DOI:10.1038/s41598-021-94603-9
PMID:34294841
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8298409/
Abstract

We aimed to develop a sandwich ELISA to detect prostate-specific membrane antigen (PSMA) on small extracellular vesicles (EVs) using T-cell immunoglobulin domain and mucin domain-containing protein 4 (Tim4) as a capture molecule for EVs and to evaluate its diagnostic potential in urologic malignancies. First, we optimized the conditions for sandwich ELISA measuring the PSMA level on EVs captured from serum by Tim4 and found that the use of highly-purified EVs released from Tim4 that had captured EVs in serum reduced the background. Second, we confirmed its validity by studying mouse xenograft model for prostate cancer (PC). Lastly, we measured PSMA-EVs in serum of patients with urologic malignancies. The PSMA-EV levels were significantly higher in metastatic PC and castration-resistant PC (CRPC) patients than in therapy-naïve PC patients. In renal cell carcinoma (RCC) patients, PSMA-EVs were elevated in those with metastasis compared with those without metastasis, which may reflect the development of the neovasculature positive for PSMA in tumors. In conclusion, we developed a sandwich ELISA for detection of PSMA-EVs using highly-purified EVs isolated from serum by Tim4. Our results suggest that PSMA-EVs may be useful to diagnose and monitor not only PC but also RCC and possibly other hypervascular solid tumors.

摘要

我们旨在开发一种使用 T 细胞免疫球蛋白结构域和粘蛋白结构域蛋白 4（Tim4）作为 EVs 捕获分子的检测前列腺特异性膜抗原（PSMA）的夹心 ELISA，用于检测小细胞外囊泡（EVs），并评估其在泌尿系统恶性肿瘤中的诊断潜力。首先，我们优化了使用 Tim4 从血清中捕获 EVs 来测量 EVs 上 PSMA 水平的夹心 ELISA 的条件，并发现使用从血清中捕获 EVs 的高度纯化的 EVs 可以减少背景。其次，我们通过研究前列腺癌（PC）的小鼠异种移植模型证实了其有效性。最后，我们测量了泌尿系统恶性肿瘤患者血清中的 PSMA-EVs。转移性 PC 和去势抵抗性 PC（CRPC）患者的 PSMA-EV 水平明显高于未经治疗的 PC 患者。在肾细胞癌（RCC）患者中，与无转移的患者相比，转移患者的 PSMA-EVs 升高，这可能反映了肿瘤中 PSMA 阳性新生血管的发展。总之，我们使用从 Tim4 从血清中分离的高度纯化的 EVs 开发了一种用于检测 PSMA-EVs 的夹心 ELISA。我们的结果表明，PSMA-EVs 可能不仅对 PC，而且对 RCC 以及可能的其他高血管实体瘤的诊断和监测都很有用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/8298409/b9fbfd06781b/41598_2021_94603_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/8298409/c53092336d02/41598_2021_94603_Fig1_HTML.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/8298409/97c77a151d7d/41598_2021_94603_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/8298409/b9fbfd06781b/41598_2021_94603_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/8298409/c53092336d02/41598_2021_94603_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/8298409/244183185979/41598_2021_94603_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/8298409/55afbcc90c8f/41598_2021_94603_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/8298409/600feecf9226/41598_2021_94603_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/8298409/a26410ab567e/41598_2021_94603_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/8298409/97c77a151d7d/41598_2021_94603_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6a0/8298409/b9fbfd06781b/41598_2021_94603_Fig7_HTML.jpg

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