Biobizkaia Health Research Institute, Barakaldo, Spain.
Institute for Cancer Research, Oslo University Hospital, Oslo, Norway.
PLoS One. 2023 Aug 1;18(8):e0289369. doi: 10.1371/journal.pone.0289369. eCollection 2023.
PTEN is a major tumor suppressor gene frequently mutated in human tumors, and germline PTEN gene mutations are the molecular diagnostic of PTEN Hamartoma Tumor Syndrome (PHTS), a heterogeneous disorder that manifests with multiple hamartomas, cancer predisposition, and neurodevelopmental alterations. A diversity of translational and splicing PTEN isoforms exist, as well as PTEN C-terminal truncated variants generated by disease-associated nonsense mutations. However, most of the available anti-PTEN monoclonal antibodies (mAb) recognize epitopes at the PTEN C-terminal tail, which may introduce a bias in the analysis of the expression of PTEN isoforms and variants. We here describe the generation and precise characterization of anti-PTEN mAb recognizing the PTEN C2-domain, and their use to monitor the expression and function of PTEN isoforms and PTEN missense and nonsense mutations associated to disease. These anti-PTEN C2 domain mAb are suitable to study the pathogenicity of PTEN C-terminal truncations that retain stability and function but have lost the PTEN C-terminal epitopes. The use of well-defined anti-PTEN mAb recognizing distinct PTEN regions, as the ones here described, will help to understand the deleterious effects of specific PTEN mutations in human disease.
PTEN 是一种重要的肿瘤抑制基因，经常在人类肿瘤中发生突变，种系 PTEN 基因突变是 PTEN 错构瘤肿瘤综合征（PHTS）的分子诊断依据，这是一种异质性疾病，表现为多种错构瘤、癌症易感性和神经发育改变。存在多种翻译和剪接的 PTEN 异构体，以及由疾病相关无义突变产生的 PTEN C 端截断变体。然而，大多数可用的抗 PTEN 单克隆抗体（mAb）识别 PTEN C 端尾部的表位，这可能会在分析 PTEN 异构体和变体的表达时引入偏差。我们在这里描述了识别 PTEN C2 结构域的抗 PTEN mAb 的产生和精确表征，以及它们用于监测与疾病相关的 PTEN 错义和无义突变的 PTEN 异构体和功能表达。这些抗 PTEN C2 结构域 mAb 适合研究保留稳定性和功能但失去了 PTEN C 端表位的 PTEN C 端截断的致病性。使用针对不同的 PTEN 区域的定义明确的抗 PTEN mAb，如本文所描述的那样，将有助于了解特定的 PTEN 突变在人类疾病中的有害影响。
