Identification of cell surface-specific markers to target human nucleus pulposus cells: Expression of carbonic anhydrase XII varies with age and degeneration
Identification of cell surface-specific markers to target human nucleus pulposus cells: Expression of carbonic anhydrase XII varies with age and degeneration
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DOI：
10.1002/art.30607
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发表时间：
2011-12-01
影响因子：
--
通讯作者：
Gallagher, William M.
中科院分区：
文献类型：
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作者：
Power, Karen A.;Grad, Sibylle;Gallagher, William M.
目的背痛是导致残疾的主要原因，影响着全世界数百万人。轴性背痛的一个原因是椎间盘髓核（NP）的变性。本研究旨在研究NP细胞与细胞表面特异性蛋白的相关性，这些蛋白不同于密切相关的细胞类型中的蛋白，即，椎间盘纤维环（AF）细胞和关节软骨（AC）软骨细胞，以鉴定用于定向递送治疗剂的潜在表面分子。我们对来自6个供体的16个人类样本进行了互补DNA微阵列分析，然后使用系统方法进行基因列表缩减。通过定量逆转录-聚合酶链反应（RT-PCR）评估了NP中比AC细胞中表达更高的基因，包含跨膜结构域，并且似乎对靶向具有吸引力。通过免疫组化和功能研究，对碳酸酐酶XII（CAXII）进行了蛋白水平的分析。微阵列结果表明AC和AF之间以及AC和NP样品之间有明显的区别。然而，AF和NP样本的转录组学特征显示出更大的受试者间相似性。在NP细胞中表达上调的552个基因中，90个含有跨膜结构域，28个通过RT-PCR定量。在年轻受试者的椎间盘髓核和退变组织中观察到最强的CAXII标记。CAXII可以被考虑用于检测或靶向退化的椎间盘细胞。此外，CAXII可能参与NP细胞的pH调节。其再生因子的定向递送潜力及其在NP细胞稳态中的功能作用值得进一步研究。
Objective Back pain is a major cause of disability, affecting millions of people worldwide. One cause of axial back pain is degeneration of the nucleus pulposus (NP) of the intervertebral disc. This study was undertaken to investigate associations of NP cells with cell surfacespecific proteins that differ from proteins in closely related cell types, i.e., intervertebral disc anulus fibrosus (AF) cells and articular cartilage (AC) chondrocytes, in order to identify potential surface molecules for directed delivery of therapeutic agents.Methods. We conducted a complementary DNA microarray analysis of 16 human samples from 6 donors, followed by gene list reduction using a systematic approach. Genes that were more highly expressed in NP than AC cells, contained transmembrane domains, and appeared attractive for targeting were assessed by quantitative reverse transcription-polymerase chain reaction (RT-PCR). As a viable candidate, carbonic anhydrase XII (CAXII) was analyzed at the protein level by immunohistochemistry and functional study.Results. Microarray results demonstrated a clear divide between the AC and AF and between the AC and NP samples. However, the transcriptomic profile of AF and NP samples displayed a greater intersubject similarity. Of the 552 genes with up-regulated expression in NP cells, 90 contained transmembrane domains, and 28 were quantified by RT-PCR. Most intense CAXII labeling was observed in the NP of discs from young subjects and in degenerative tissue.Conclusion. CAXII may be considered for detection or targeting of degenerating disc cells. Furthermore, CAXII may be involved in pH regulation of NP cells. Its potential for directed delivery of regenerative factors and its functional role in NP cell homeostasis warrant further investigation.