Department of Chemistry, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Department of Molecular Medicine, The Scripps Research Institute, La Jolla, CA 92037, USA.
Sci Transl Med. 2017 Sep 13;9(407). doi: 10.1126/scitranslmed.aam7621.
Increasing evidence supports the hypothesis that soluble misfolded protein assemblies contribute to the degeneration of postmitotic tissue in amyloid diseases. However, there is a dearth of reliable nonantibody-based probes for selectively detecting oligomeric aggregate structures circulating in plasma or deposited in tissues, making it difficult to scrutinize this hypothesis in patients. Hence, understanding the structure-proteotoxicity relationships driving amyloid diseases remains challenging, hampering the development of early diagnostic and novel treatment strategies. We report peptide-based probes that selectively label misfolded transthyretin (TTR) oligomers circulating in the plasma of TTR hereditary amyloidosis patients exhibiting a predominant neuropathic phenotype. These probes revealed that there are much fewer misfolded TTR oligomers in healthy controls, in asymptomatic carriers of mutations linked to amyloid polyneuropathy, and in patients with TTR-associated cardiomyopathies. The absence of misfolded TTR oligomers in the plasma of cardiomyopathy patients suggests that the tissue tropism observed in the TTR amyloidoses is structure-based. Misfolded oligomers decrease in TTR amyloid polyneuropathy patients treated with disease-modifying therapies (tafamidis or liver transplant-mediated gene therapy). In a subset of TTR amyloid polyneuropathy patients, the probes also detected a circulating TTR fragment that disappeared after tafamidis treatment. Proteomic analysis of the isolated TTR oligomers revealed a specific patient-associated signature composed of proteins that likely associate with the circulating TTR oligomers. Quantification of plasma oligomer concentrations using peptide probes could become an early diagnostic strategy, a response-to-therapy biomarker, and a useful tool for understanding structure-proteotoxicity relationships in the TTR amyloidoses.
越来越多的证据支持这样一种假说,即可溶性错误折叠蛋白聚集体有助于淀粉样变性疾病中处于有丝分裂后组织的退化。然而,目前缺乏可靠的非抗体探针,无法选择性地检测在血浆中循环或在组织中沉积的寡聚聚集结构,这使得难以在患者中仔细研究这一假说。因此,理解导致淀粉样变性疾病的结构-毒性关系仍然具有挑战性,这阻碍了早期诊断和新型治疗策略的发展。我们报告了基于肽的探针,这些探针可选择性地标记在 TTR 遗传性淀粉样变性患者的血浆中循环的错误折叠转甲状腺素蛋白(TTR)寡聚体,这些患者表现出主要的神经病变表型。这些探针表明,在健康对照者、与淀粉样多神经病相关突变的无症状携带者以及 TTR 相关性心肌病患者中,错误折叠的 TTR 寡聚体要少得多。在心肌病患者的血浆中不存在错误折叠的 TTR 寡聚体表明,在 TTR 淀粉样变性中观察到的组织嗜性是基于结构的。用疾病修饰疗法(塔法米地或肝移植介导的基因治疗)治疗 TTR 淀粉样多神经病患者后,TTR 寡聚体减少。在 TTR 淀粉样多神经病患者的亚组中,探针还检测到一种循环 TTR 片段,该片段在塔法米地治疗后消失。对分离的 TTR 寡聚体进行蛋白质组学分析揭示了一种特定的与患者相关的特征,其组成部分是可能与循环 TTR 寡聚体结合的蛋白质。使用肽探针定量检测血浆寡聚体浓度可能成为一种早期诊断策略、一种治疗反应生物标志物,以及一种用于理解 TTR 淀粉样变性中结构-毒性关系的有用工具。