Department of Chemistry, W. M. Keck Institute of Proteomics Technology and Applications, The George Washington University, Washington, D. C., USA.
PLoS One. 2010 Sep 7;5(9):e12590. doi: 10.1371/journal.pone.0012590.
Viral transformation of a cell starts at the genetic level, followed by changes in the proteome and the metabolome of the host. There is limited information on the broad metabolic changes in HTLV transformed cells.
Here, we report the detection of key changes in metabolites and lipids directly from human T-lymphotropic virus type 1 and type 3 (HTLV1 and HTLV3) transformed, as well as Tax1 and Tax3 expressing cell lines by laser ablation electrospray ionization (LAESI) mass spectrometry (MS). Comparing LAESI-MS spectra of non-HTLV1 transformed and HTLV1 transformed cells revealed that glycerophosphocholine (PC) lipid components were dominant in the non-HTLV1 transformed cells, and PC(O-32:1) and PC(O-34:1) plasmalogens were displaced by PC(30:0) and PC(32:0) species in the HTLV1 transformed cells. In HTLV1 transformed cells, choline, phosphocholine, spermine and glutathione, among others, were downregulated, whereas creatine, dopamine, arginine and AMP were present at higher levels. When comparing metabolite levels between HTLV3 and Tax3 transfected 293T cells, there were a number of common changes observed, including decreased choline, phosphocholine, spermine, homovanillic acid, and glycerophosphocholine and increased spermidine and N-acetyl aspartic acid. These results indicate that the lipid metabolism pathway as well as the creatine and polyamine biosynthesis pathways are commonly deregulated after expression of HTLV3 and Tax3, indicating that the noted changes are likely due to Tax3 expression. N-acetyl aspartic acid is a novel metabolite that is upregulated in all cell types and all conditions tested.
We demonstrate the high throughput in situ metabolite profiling of HTLV transformed and Tax expressing cells, which facilitates the identification of virus-induced perturbations in the biochemical processes of the host cells. We found virus type-specific (HTLV1 vs. HTLV3), expression-specific (Tax1 vs. Tax3) and cell-type-specific (T lymphocytes vs. kidney epithelial cells) changes in the metabolite profiles. The new insight on the affected metabolic pathways can be used to better understand the molecular mechanisms of HTLV induced transformation, which in turn can result in new treatment strategies.
病毒对细胞的转化始于遗传水平，随后是宿主蛋白质组和代谢组的变化。关于 HTLV 转化细胞的广泛代谢变化，信息有限。
在这里，我们报告了通过激光烧蚀电喷雾电离（LAESI）质谱（MS）直接从人 T 淋巴细胞白血病病毒 1 型和 3 型（HTLV1 和 HTLV3）以及 Tax1 和 Tax3 表达细胞系中检测到关键代谢物和脂质变化。比较非 HTLV1 转化细胞和 HTLV1 转化细胞的 LAESI-MS 谱表明，甘油磷酸胆碱（PC）脂质成分在非 HTLV1 转化细胞中占主导地位，而 HTLV1 转化细胞中 PC（O-32：1）和 PC（O-34：1）类质体被 PC（30：0）和 PC（32：0）取代。在 HTLV1 转化细胞中，胆碱、磷酸胆碱、精脒和谷胱甘肽等下调，而肌酸、多巴胺、精氨酸和 AMP 水平升高。比较 HTLV3 和 Tax3 转染 293T 细胞之间的代谢物水平时，观察到许多共同变化，包括胆碱、磷酸胆碱、精脒、高香草酸和甘油磷酸胆碱减少，以及精脒和 N-乙酰天冬氨酸增加。这些结果表明，在表达 HTLV3 和 Tax3 后，脂质代谢途径以及肌酸和多胺生物合成途径普遍失调，表明所注意到的变化可能是由于 Tax3 表达所致。N-乙酰天冬氨酸是一种新型代谢物，在所有测试的细胞类型和条件下均上调。
我们展示了 HTLV 转化和 Tax 表达细胞的高通量原位代谢物分析，这有助于确定病毒对宿主细胞生化过程的诱导扰动。我们发现病毒类型特异性（HTLV1 与 HTLV3）、表达特异性（Tax1 与 Tax3）和细胞类型特异性（T 淋巴细胞与肾上皮细胞）代谢谱变化。受影响代谢途径的新见解可用于更好地理解 HTLV 诱导转化的分子机制，进而可以产生新的治疗策略。
